From 776af5504e8a7c88bc2cb9a2524dbcd1bd044c4d Mon Sep 17 00:00:00 2001 From: Keoni Gandall Date: Wed, 28 Aug 2024 21:35:30 -0700 Subject: [PATCH 1/9] init --- go.work | 1 + parts/README.md | 33 ++++ parts/go.mod | 10 ++ parts/go.sum | 6 + parts/linkers.md | 244 ++++++++++++++++++++++++++ parts/main.go | 130 ++++++++++++++ parts/parts/bsub.yaml | 282 +++++++++++++++++++++++++++++++ parts/parts/ecoli/ecoli.yaml | 103 +++++++++++ parts/parts/ecoli/origins.yaml | 52 ++++++ parts/parts/ecoli/promoters.yaml | 101 +++++++++++ parts/parts/pichia_toolkit.yaml | 114 +++++++++++++ parts/parts/vnat.yaml | 178 +++++++++++++++++++ parts/parts/yeast_toolkit.yaml | 252 +++++++++++++++++++++++++++ parts/primers.tsv | 166 ++++++++++++++++++ 14 files changed, 1672 insertions(+) create mode 100644 parts/README.md create mode 100644 parts/go.mod create mode 100644 parts/go.sum create mode 100644 parts/linkers.md create mode 100644 parts/main.go create mode 100644 parts/parts/bsub.yaml create mode 100644 parts/parts/ecoli/ecoli.yaml create mode 100644 parts/parts/ecoli/origins.yaml create mode 100644 parts/parts/ecoli/promoters.yaml create mode 100644 parts/parts/pichia_toolkit.yaml create mode 100644 parts/parts/vnat.yaml create mode 100644 parts/parts/yeast_toolkit.yaml create mode 100644 parts/primers.tsv diff --git a/go.work b/go.work index 1c851ee..0509257 100644 --- a/go.work +++ b/go.work @@ -4,4 +4,5 @@ use ( ./external ./lib ./py + ./parts ) diff --git a/parts/README.md b/parts/README.md new file mode 100644 index 0000000..9429ecc --- /dev/null +++ b/parts/README.md @@ -0,0 +1,33 @@ +# KG Parts Library + +This is the KG Genetic Parts library. It is written with basic yaml for data portability purposes. + +The library is static and intended for embedded distribution in python or go, or online using json. The built files in json or yaml are therefore saved to the project itself. + +## Purpose + +The KG Genetic Parts Library is a complete genetic parts library, covering all major organisms, with consistent and high-quality documentation. + +## Rules +1. All genes must have a unique name, including genes used between different toolkits +2. Parts should be accessible from a URL appended with `.json`. No weird special characters or spaces. +3. A given sequence should only have 1 name. +4. In cases of proteins or tags encoded for a certain organism, add a parathesis tag to the end of the protein. For example, `SceI(Scerevisae)`. +5. Sequences are identified by their fragment seqhash. Any code interacting with these genetic parts should identify by seqhash, NOT name. The name is only for human readability sake. +6. No I-SceI sites or enzyme expression. + +## Some more +- terminators have stop codons (`ATCCTAA` prefix) in the case you want to use a c-taggable protein with a GS on the c terminal. ctag terminators rely on ctags to have stop codons. +- Unlike most MoClo toolkits, we do not place the promoter next to the coding sequence in eukaryotic toolkits. There is space for kozak sequences or ntags, which occupy the same prefix/suffix space. + +## Primers +- Dialout primers P1-P96 are reserved for synthesis and assembly usage. Do not use them. +- Dialout primers P97-P144 are reserved for pooling toolkits that are going to be distributed together, so try not to use them. +- Dialout primers 165-161 are used for standardized primers. Do not use them. +- If you need to use primers, it is recommended to use set P150,P151,P152,P153,P154,P155,P156,P158 + +## Organisms: +- Escherichia coli (Ec) +- Bacillus subtilis (Bs) +- Vibrio natriegens (Vn) +- Saccharomyces cerevisiae (Sc) diff --git a/parts/go.mod b/parts/go.mod new file mode 100644 index 0000000..181d68d --- /dev/null +++ b/parts/go.mod @@ -0,0 +1,10 @@ +module github.com/koeng101/dnadesign/parts + +go 1.22.5 + +require ( + github.com/koeng101/dnadesign/lib v0.0.0-00010101000000-000000000000 + gopkg.in/yaml.v2 v2.4.0 +) + +replace github.com/koeng101/dnadesign/lib => ../lib diff --git a/parts/go.sum b/parts/go.sum new file mode 100644 index 0000000..9d687d8 --- /dev/null +++ b/parts/go.sum @@ -0,0 +1,6 @@ +github.com/koeng101/dnadesign/lib v0.0.0-20240829011445-cf75dc91ccf1 h1:Yxg8tE7B2suXwpK2qGVxNxZ5zIW4r2AeI3quxq3/UOQ= +github.com/koeng101/dnadesign/lib v0.0.0-20240829011445-cf75dc91ccf1/go.mod h1:oTFawZ8aVIhAQSRNJKH9Yk4rp/nTDBVLDSsYEaxJPTQ= +gopkg.in/check.v1 v0.0.0-20161208181325-20d25e280405 h1:yhCVgyC4o1eVCa2tZl7eS0r+SDo693bJlVdllGtEeKM= +gopkg.in/check.v1 v0.0.0-20161208181325-20d25e280405/go.mod h1:Co6ibVJAznAaIkqp8huTwlJQCZ016jof/cbN4VW5Yz0= +gopkg.in/yaml.v2 v2.4.0 h1:D8xgwECY7CYvx+Y2n4sBz93Jn9JRvxdiyyo8CTfuKaY= +gopkg.in/yaml.v2 v2.4.0/go.mod h1:RDklbk79AGWmwhnvt/jBztapEOGDOx6ZbXqjP6csGnQ= diff --git a/parts/linkers.md b/parts/linkers.md new file mode 100644 index 0000000..4cb98b4 --- /dev/null +++ b/parts/linkers.md @@ -0,0 +1,244 @@ +**NOTE! This is old documentation on linkers, which is still relevant for part overhangs, but the linkers themselves are still under design.** + +# Linkers + +There have been many standard methods for assemblying DNA, with the most notable being the [BioBrick assembly](https://en.wikipedia.org/wiki/BioBrick) method being developed in 2003. However, BioBrick assembly could only assemble 2 DNA parts at once, and therefore limited the amount of assembly that one could do in a short amount of time. In 2008, a new method called [GoldenGate assembly](https://dx.doi.org/10.1371%2Fjournal.pone.0003647) was developed that overcame limitations of BioBrick assembly, allowing many fragments to be put together at once in a single tube. In 2011, this GoldenGate assembly was standardized with the [MoClo assembly](https://doi.org/10.1371/journal.pone.0016765) method. + +The Linkers Collection is a collection of linkers ([here](https://doi.org/10.3389/fbioe.2019.00271) is a good introduction) for MoClo assembly. These linkers have 2 special attributes: + +- The BsaI overhangs are optimized using [empirical data](https://doi.org/10.1371/journal.pone.0238592) +- The assembly is recursive - the same linkers are used at each level of assembly. This is accomplished using methyltransferases which [methylate at GACNNNGTC](https://doi.org/10.1093%2Fnar%2Fgky596) or [methylate at CCGG positions](http://www.greatlakesbiotech.org/news/2016/8/26/designing-a-low-cost-molecular-biology-platform) + +### Changes +Linkers were recreated from my previous toolkits because `CGAG,GTCT` are required for the assembly method, and these greatly lowered the efficiency of traditional MoClo, mainly because GGAG is used as one of the classic overhangs. I added `AAAA` and `GGGG` as two standard overhangs - while they aren't used in in normal assemblies, they are available for if a fragment needs to build polyA/polyT/polyG/polyC. + +overhangs: `CGAG,GTCT,GGGG,AAAA,AACT,AATG,ATCC,CGCT,TTCT,AAGC,ATAG,ATTA,ATGT,ACTC,ACGA,TATC,TAGG,TACA,TTAC,TTGA,TGGA,GAAG,GACC,GCCG` +overhangs with numbers: +1. CGAG +2. TACA +3. AACT +4. AATG +5. ATCC +6. CGCT +7. GTCT +8. AAGC +9. ATAG +10. ATTA +11. TTCT +12. ATGT +13. ACTC +14. ACGA +15. TATC +16. TAGG +17. TACA +18. TTAC +19. TTGA +20. TGGA +21. GAAG +22. GACC +23. GCCG +24. AAAA +25. GGGG + +Once a plasmid cloned using a linker, its new definition is derived from the particular linkers used to construct it. For example, an assembly with A1(2) + B1(4) creates a new plasmid with a fragment defined with the overhangs 2 and 4. + +Note: 11 is no longer defined as ntag. + +# Normal build process +### Simple build +A simple build constructs an Escherichia coli vector with no added fluff. +* [A1] CGAG - TACA (linker prefix) +* [P] TACA - AACT (promoter) +* [R] AACT - AATG (rbs) +* [C] AATG - ATCC (cds) +* [T] ATCC - CGCT (terminator) +* [B1] CGCT - GTCT (linker suffix) +* [E1] GTCT - CGAG (e coli vector) + +Or, with numbered overhangs: + +``` +Simple build: +1 [linker prefix] 2 [promoter] 3 [RBS] 4 [CDS] 5 [terminator] 6 [linker suffix] 7 [vector1] ... +``` + +### Operon assembly +The first operon component, X number of mid operon components, and the last operon component can then be combined based off of their prefix and suffix linkers. + +Operon assembly first: +* [A1] CGAG - TACA (linker prefix) +* [P] TACA - AACT (promoter) +* [R] AACT - AATG (rbs) +* [C] AATG - ATCC (cds) +* [B2] ATCC - GTCT (linker suffix) +* [E1] GTCT - CGAG (e coli vector) + +Operon assembly mid: +* [A2] CGAG - AACT (linker prefix) +* [R] AACT - AATG (rbs) +* [C] AATG - ATCC (cds) +* [B2] ATCC - GTCT (linker suffix) +* [E1] GTCT - CGAG (e coli vector) + +Operon assembly last: +* [A2] CGAG - AACT (linker prefix) +* [R] AACT - AATG (rbs) +* [C] AATG - ATCC (cds) +* [T] ATCC - CGCT (terminator) +* [B1] CGCT - GTCT (linker suffix) +* [E1] GTCT - CGAG (e coli vector) + +``` +Operon assembly (first): +1 [linker prefix] 2 [promoter] 3 [RBS] 4 [CDS] 5 [linker suffix] 7 [vector1] ... + +Operon assembly (mid): +1 [linker prefix] 3 [RBS] 4 [CDS] 5 [linker suffix] 7 [vector1] ... + +Operon assembly (last): +1 [linker prefix] 3 [RBS] 4 [CDS] 5 [terminator] 6 [linker suffix] 7 [vector1] ... +``` + + +### Shuttle vector +Often, users will want to move transcription units to new organisms of interest. The following is a simple shuttle vector: + +* [A] CGAG - TACA (linker prefix) +* [P] TACA - AACT (promoter) +* [R] AACT - AATG (rbs) +* [C] AATG - ATCC (cds) +* [T] ATCC - CGCT (terminator) +* [B] CGCT - GTCT (linker suffix) +* [D] GTCT - AAGC (target origin of replication) +* [S] AAGC - ATAG (target selective marker) +* [E2] ATAG - CGAG (e coli vector 2) + +``` +Simple shuttle: +1 [linker prefix] 2 [promoter] 3 [RBS] 4 [CDS] 5 [terminator] 6 [linker suffix] 7 [target ori] 8 [target marker] 9 [vector2] ... +``` + +### Integration vector +Instead of shuttle vectors, users will sometimes want to integrate sections of DNA into their organism of interest. The following is a build definition for an integration vector: + +* [A] CGAG - TACA (linker prefix) +* [P] TACA - AACT (promoter) +* [R] AACT - AATG (rbs) +* [C] AATG - ATCC (cds) +* [T] ATCC - GTCT (terminator) +* [B] GTCT - ATAG (linker suffix) +* [D] GTCT - AAGC (target ori / downstream homology) +* [S] AAGC - ATAG (target selective marker) +* [EC3] ATAG - ATTA (e coli vector 3) +* [U] ATTA - CGAG (upstream homology) +``` +Integration vector: +1 [linker prefix] 2 [promoter] 3 [RBS] 4 [CDS] 5 [terminator] 6 [linker suffix] 7 [downstream homology] 8 [target marker] 9 [vector3] 10 [upstream homology] ... +``` + +### Protein tags +It is common that one would want to add tags to a protein sequence. You can add to the N terminal or C terminal. The N tag is carried along with the ribosomal binding site. + +* [A1] CGAG - TACA (linker prefix) +* [P] TACA - AACT (promoter) +* [R] AACT - AATG (rbs/nterminal tag) +* [C] AATG - ATCC (cds) +* [Cc] ATCC - ATGT (c terminal tag) +* [Tt] ATGT - CGCT (terminator with c terminal tag) +* [B1] CGCT - GTCT (linker suffix) +* [E1] GTCT - CGAG (e coli vector) + +Or, with numbered overhangs: + +``` +Protein tag build: +1 [linker prefix] 2 [promoter] 3 [RBS/N tag] 4 [CDS] 5 [C tag] 12 [terminator] 6 [linker suffix] 7 [vector1] ... +``` + +### Conserved overhangs +The following overhangs are reserved for more complicated backbone assemblies when manipulating Escherichia coli backbones. These represent overhangs 13, 14, 15, 16. + +* [EFX] NNNN - ACTC (e coli vector compatibilizer prefix) +* [M] ACTC - ACGA (e coli marker) {always in R6K backbones} +* [O] ACGA - TATC (e coli origin) +* [Z] TATC - TAGG (package signal, usually oriT) +* [ERX] TAGG - NNNN (e coli vector compatibilizer suffix) + +EFX and ERX can have the following definitions: +* EF1 +* EF2 +* ER1 +* ER3 + +An E3 vector, for example, would use the flanks EF2 and ER3, while E1 would use EF1 and ER1. + +Backbones parts are typically used in situations where users can't rely on ccdB counter-selection normally present in cached vectors - for example, when constructing a new vector that a single part, like a CDS, can be integrated into. + +In addition, the following overhangs are used for specialty vector construction: +* TTAC +* TTGA + +# Special constructions +### Backbone part construction +Parts can be constructed to function in backbones, except type M parts. +``` +1 [linker prefix (7)] 2 [promoter] 3 [RBS] 4 [Kanamycin resistance coding sequence] 5 [terminator] 6 [linker suffix (8)] 7 [negative selection marker] 8... [vector] ... +After assembly: +7 [Kanamycin resistance] 13 +``` + +### Reversing during higher level construction +Each linker is defined with a number, representing the overhang it switches its assembly to. These can be negative numbers, which are the reverse complements of that particular overhang. These can be used to flip sequences during construction. For example: +``` +Simplified transcription unit now represented by `-->` + +1( ----> )2 + -3( --> )-2 + 3( -> )4 = ( ----> <-- -> ) +``` + +## Vectors +### Vector [E0] construction +E0 vectors, or vectors used in recurse builds or normal foundry synthesis orders, are always constructs by-hand in a non-modular fashion using BbsI. Users cannot construct E0 vectors in our foundry. + +### Resistance marker construction (M) +Escherichia coli resistrction marker parts, or M parts, are always in R6K vectors. This allows switching to a non-R6K strain as a way to select out the original vector. The foundry does provide R6K transformation resources. + +### Vector [E1,E2,E3] construction +Vector types E1,E2,E3 are used in constructing normal DNA. They use ccdB for negative selection. They are always constructed from (M) parts, so that the marker can be switched during the GoldenGate reaction. + +### Specialty vector construction +Speciality vectors use special linkers to add a `ccdB-MOsp87` cassette into any site of a normal construction. For example, if you have an expression vector that you know works well, you can swap out your gene of interest with 2 linkers and `ccdB-MOsp87`. This creates a new vector that you can directly add new genes of interest into, without adding the corresponding promoters or terminators to the reaction. + +### Recurse builds +Recurse builds are the exception case to the rule that assemblies are redefined with their given linkers. In recurse builds, nothing is ever redefined: whatever overhang the input fragments had, the output assembly will have. + +A recursive build is designed to build DNA from blocks. The designed DNA can be built from any number of blocks at any given step: If there are 20 blocks to be added together, one could design the DNA to be built with 4-5, or 2-2-5, or anything else. For difficult sequences, this allows clone-time optimizations, without going back to the synthesis phase. +* [A1] CGAG - {} (recurse linker prefix) +* [ ] {} - {} (fragment of interest) +* [B1] {} - GTCT (recurse linker suffix) +* [E1] GTCT - CGAG (e coli vector) + +``` +recurse build: +1 [recurse linker prefix] x [n] x [n+1...] x [recurse linker suffix] 7 [vector1] ... +``` + + +# FAQ +## How were the linkers designed +The efficiency designer is based off of [this datasheet](https://doi.org/10.1371/journal.pone.0238592.s001) from "Enabling one-pot Golden Gate assemblies of unprecedented complexity using data-optimized assembly design". I am using [Poly](https://github.com/timothystiles/poly)'s fragment designer, which I coded. + +## What are linkers? +When building a construct using GoldenGate, simply ligate linkers between your vector and your genes during a GoldenGate reaction to enable use of that gene in multi-gene constructs. + +Typically, you will do an assembly reaction (also known as a level 1 cloning reaction in MoClo lingo) to give context to your gene. For example, you may have a protein called GFP that you wish to express. In this case, you would do a level 1 cloning reaction to contextualize GFP with a proper promoter and terminator for your target organism to make a transcriptional unit (TU). In that reaction, you may have to add linkers to connect your construct into the vector it belongs in. + +Afterwards, you can combine the GFP transcriptional unit to up to 24 other constructs with clever usage of linkers. To answer specifically which ones to use and when, read below. + +## What linkers are included in the Linkers Collection? + +This collection has 384 linkers. There are 96 linkers for building independent transcription units, 96 linkers for building operons, 96 linkers for recursive builds, and 96 linkers for defining new speciality vectors. For each set of 96, linkers are split into 48 prefix and suffix linkers. Those 48 linkers are split into 24 positive and 24 negative linkers. The positive linkers are used for constructing genes in the forward direction, and the negative linkers are used to construct genes in the reverse direction. + +Each overhang is assigned a number. The reverse complement of each overhang is represented as the negative version of its number, which is also how we are able to flip constructs. + +Linkers are named with a 1 letter + 3 number scheme, separated by underscores ( _ ) in the format `Y_X_X_X`. Y describes the direction (F, or forward, for prefix, and R, or reverse, for suffix) of the linker. The first 2 numbers describe the two overhangs which the linker itself will be cut out with, the third number describes the overhang which the linker will introduce to the construct. After a GoldenGate assembly and transformation, anything between the prefix and suffix lin +kers can be cut out with BsaI and used in another assembly. diff --git a/parts/main.go b/parts/main.go new file mode 100644 index 0000000..226f505 --- /dev/null +++ b/parts/main.go @@ -0,0 +1,130 @@ +package main + +import ( + "embed" + "encoding/json" + "fmt" + "io/fs" + "io/ioutil" + "os" + "path/filepath" + "strings" + + "github.com/koeng101/dnadesign/lib/seqhash" + "gopkg.in/yaml.v2" +) + +// Embed the entire parts directory +// +//go:embed parts +var embeddedFiles embed.FS + +// Part represents a single part part. +type Part struct { + Seqhash string `yaml:"seqhash" json:"seqhash"` + Name string `yaml:"name" json:"name"` + Description string `yaml:"description" json:"description"` + Tags []string `yaml:"tags" json:"tags"` + Prefix string `yaml:"prefix" json:"prefix"` + Suffix string `yaml:"suffix" json:"suffix"` + Sequence string `yaml:"sequence" json:"sequence"` +} + +func main() { + // Use fs.WalkDir to walk through embedded directory + partMap := make(map[string]Part) + err := fs.WalkDir(embeddedFiles, "parts", func(path string, d fs.DirEntry, err error) error { + if err != nil { + fmt.Printf("Error walking through embedded directory: %v\n", err) + return err + } + + if !d.IsDir() && filepath.Ext(path) == ".yaml" { + data, err := embeddedFiles.ReadFile(path) + if err != nil { + fmt.Printf("Error reading embedded file %s: %v\n", path, err) + return err + } + + var contents map[string]Part + err = yaml.Unmarshal(data, &contents) + if err != nil { + fmt.Printf("Error unmarshalling YAML from embedded file %s: %v\n", path, err) + return err + } + + for name, part := range contents { + if part.Prefix == "" || part.Suffix == "" || part.Sequence == "" { + continue + } + sq, err := seqhash.EncodeHash2(seqhash.Hash2Fragment(strings.ToUpper(part.Prefix+part.Sequence+part.Suffix), 4, 4)) + if err != nil { + fmt.Printf("Error seqhashing: %v\n", err) + return err + } + partMap[sq] = Part{Name: name, Seqhash: sq, Description: part.Description, Tags: part.Tags, Prefix: strings.ToUpper(part.Prefix), Suffix: strings.ToUpper(part.Suffix), Sequence: strings.ToUpper(part.Sequence)} + } + } + return nil + }) + if err != nil { + fmt.Printf("Error walking the embedded file system: %v\n", err) + } + + /* + Build parts directory + */ + + directory := "build/parts" + + // Ensure the directory exists + if _, err := os.Stat(directory); os.IsNotExist(err) { + os.MkdirAll(directory, 0755) + } + + // Clear all files in the directory + files, err := ioutil.ReadDir(directory) + if err != nil { + fmt.Println("Error reading directory:", err) + return + } + for _, file := range files { + err := os.RemoveAll(filepath.Join(directory, file.Name())) + if err != nil { + fmt.Println("Error removing file:", err) + return + } + } + // Serialize and write files for each part + for _, part := range partMap { + fmt.Println(part) + jsonFileName := filepath.Join(directory, part.Seqhash+".json") + yamlFileName := filepath.Join(directory, part.Seqhash+".yaml") + + // Marshal part to JSON + jsonData, err := json.MarshalIndent(part, "", " ") + if err != nil { + fmt.Println("Error marshaling to JSON:", err) + continue + } + + // Marshal part to YAML + yamlData, err := yaml.Marshal(part) + if err != nil { + fmt.Println("Error marshaling to YAML:", err) + continue + } + + // Write JSON file + if err := ioutil.WriteFile(jsonFileName, jsonData, 0644); err != nil { + fmt.Println("Error writing JSON file:", err) + continue + } + + // Write YAML file + if err := ioutil.WriteFile(yamlFileName, yamlData, 0644); err != nil { + fmt.Println("Error writing YAML file:", err) + continue + } + } +} diff --git a/parts/parts/bsub.yaml b/parts/parts/bsub.yaml new file mode 100644 index 0000000..6b3bdfa --- /dev/null +++ b/parts/parts/bsub.yaml @@ -0,0 +1,282 @@ +PxylA: + description: Xylose inducible promoter + prefix: TACA + suffix: AACT + tags: [promoter] + sequence: aaggccaaaaaactgctgccttcggatcagcgatatccacttcatccactccatttgtttaatctttaaattaagtatcaacatagtacatagcgaatcttccctttattatatctaatgtgttcataaaaaactaaaaaaaatattgaaaatactgacgaggttatataagatgaaaataagttagtttgtttaaacaacaaactaataggtgatgtacttactatatgaaataaaatgcatctgggatcccaagcttatcgaa +PliaI: + description: Bacitracin-inducible promoter + prefix: TACA + suffix: AACT + tags: [promoter] + sequence: attggccaaagcagaaaggtccgacctaattaaagaaagggaagcaagtgttcatctgtaaagggttttaaaacgccatgcctcgtgcatggcgtttttttgtgccaatgggtccggtgcgagatacgactccggtcttatataaaaatcaatctctgattcgttttgcatatcttccaacttgtataagatgaagacaaggaaaacga +P_J23101: + description: Very weak constitutive promoter (Bsub) + prefix: TACA + suffix: AACT + tags: [promoter] + sequence: tttacagctagctcagtcctaggtattatgctagc +PlepA: + description: Strong constitutive promoter + prefix: TACA + suffix: AACT + tags: [promoter] + sequence: agtcaatgtatgaatggatacgggatatgaatcaataagtacgtgaaagagaaaagcaacccagatatgatagggaacttttctctttcttgttttacattgaatctttacaatcctattgatataatctaagctagtgtattttgcgtttaatagt +PliaG: + description: Constitutive promoter + prefix: TACA + suffix: AACT + tags: [promoter] + sequence: caaaaatcagaccagacaaaagcggcaaatgaataagcggaacggggaaggatttgcggtcaagtccttcccttccgcacgtatcaattcgcaagcttttcctttataatagaatgaatga +Pveg: + description: Very strong constitutive promoter + prefix: TACA + suffix: AACT + tags: [promoter] + sequence: ggagttctgagaattggtatgccttataagtccaattaacagttgaaaacctgcataggagagctatgcgggttttttattttacataatgatacataatttaccgaaacttgcggaacataattgaggaatcatagaattttgtcaaaataattttattgacaacgtcttattaacgttgatataatttaaattttatttgacaaaaatgggctcgtgttgtacaataaatgtagt +US_amyE: + description: upstream flank amyE + prefix: ATTA + suffix: CGAG + tags: [upstream_flank] + sequence: ttaacaaaattctccagtcttcacatcggtttgaaaggaggaagcggaagaatgaagtaagagggatttttgactccgaagtaagtcttcaaaaaatcaaataaggagtgtcaagaatgtttgcaaaacgattcaaaacctctttactgccgttattcgctggatttttattgctgtttcatttggttctggcaggaccggcggctgcgagtgctgaaacggcgaacaaatcgaatgagcttacagcaccgtcgatcaaaagcggaaccattcttcatgcatggaattggtcgttcaatacgttaaaacacaatatgaaggatattcatgatgcaggatatacagccattcagacatctccgattaaccaagtaaaggaagggaatcaaggagataaaagcatgtcgaactggtactggctgtatcagccgacatcgtatcaaattggcaaccgttacttaggtactgaacaagaatttaaagaaatgtgtgcagccgctgaagaatatggcataaaggtcattgttgacgcggtcatcaatcataccaccagtgattatgccgcgatttccaatgaggttaagagtattccaaactggacacatggaaacacacaaattaaaaactggtctgatc +DS_amyE: + description: downstream flank amyE + prefix: GTCT + suffix: AAGC + tags: [downstream_flank] + sequence: agagtcgacatggatgagcgatgatgatatccgtttaggctgggcggtgatagcttctcgttcaggcagtacgcctcttttcttttccagacctgagggaggcggaaatggtgtgaggttcccggggaaaagccaaataggcgatcgcgggagtgctttatttgaagatcaggctatcactgcggtcaatagatttcacaatgtgatggctggacagcctgaggaactctcgaacccgaatggaaacaaccagatatttatgaatcagcgcggctcacatggcgttgtgctggcaaatgcaggttcatcctctgtctctatcaatacggcaacaaaattgcctgatggcaggtatgacaataaagctggagcgggttcatttcaagtgaacgatggtaaactgacaggcacgatcaatgccaggtctgtagctgtgctttatcctgatgatattgcaaaagcgcctcatgttttccttgagaattacaaaacaggtgtaacacattctttcaatgatcaactgacgattaccttgcgtgcagatgcgaatacaacaaaagccgtttatcaaatcaataatggaccagagacggcgtttaaggatggagatcaattcacaatcggaaaaggagatccatttggcaaaacatacaccatcatgttaaaaggaacgaacagtgatggtgtaacgaggaccgagaaatacagttttgttaaaagagatccagcgtcggccaaaaccatcggctatcaaaatccgaatcattggagccaggtaaatgcttatatctataaacatgatgggagccgagtaattgaattgaccggatcttggcctggaaaaccaatgactaaaaatgcagacggaatttacacgctgacgctgcctgcggacacggatacaaccaacgcaaaagtgatttttaataatggcagcgcccaagtgcccggtcagaatcagcctggctttgattacgtgctaaatggtttatataatgactcgggcttaagcggttctcttccccattgagggcaaggctagacgggacttaccgaaagaaaccatcaatgatggtttcttttttgttcataaatcagacaaaacttttctcttgcaaaagtttgtgaagtgttgcacaatataaatgtgaaatacttcacaaacaaaaagacatcaaagagaaacataccctg +camR(Bsub): + description: Chloramphenicol resistance in Bacillus subtilis + prefix: AAGC + suffix: ATAG + tags: [selection] + sequence: tccggcaaaaaagggcaaggtgtcaattctcatgtttgacagcttatcatcggcaatagttacccttattatcaagataagaaagaaaaggatttttcgctacgctcaaatcctttaaaaaaacacaaaagaccacattttttaatgtggtctttattcttcaactaaagcacccattagttcaacaaacgaaaattggataaagtgggatatttttaaaatatatatttatgttacagtaatattgacttttaaaaaaggattgattctaatgaagaaagcagacaagtaagcctcctaaattcactttagataaaaatttaggaggcatatcaaatgaactttaataaaattgatttagacaattggaagagaaaagagatatttaatcattatttgaaccaacaaacgacttttagtataaccacagaaattgatattagtgttttataccgaaacataaaacaagaaggatataaattttaccctgcatttattttcttagtgacaagggtgataaactcaaatacagcttttagaactggttacaatagcgacggagagttaggttattgggataagttagagccactttatacaatttttgatggtgtatctaaaacattctctggtatttggactcctgtaaagaatgacttcaaagagttttatgatttatacctttctgatgtagagaaatataatggttcggggaaattgtttcccaaaacacctatacctgaaaatgctttttctctttctattattccatggacttcatttactgggtttaacttaaatatcaataataatagtaattaccttctacccattattacagcaggaaaattcattaataaaggtaattcaatatatttaccgctatctttacaggtacatcattctgtttgtgatggttatcatgcaggattgtttatgaactctattcaggaattgtcagataggcctaatgactggcttttataatatgagataatgccgactgtactttttacagtcggttttctaatgtcactaacctgccccgttagttgaagaaggtttttatattacagctccagatcctctacgccggacgcatcgtggccggcatcaccggcgccacaggtgcggttgctggcgcctatatcgccgacatcaccgatggggaagatcgggctcgccacttcgggctcatgagcgcttgtttcggcgtgggtatggtggcaggccccgtggccgggggactgttgggcgccatctccttgcatgcaccattccttgcggcggcggtgctcaacggcctcaacctactactgggctgcttcctaatgcaggagtcgcataagggag +US_lacA: + description: upstream flank lacA + prefix: ATTA + suffix: CGAG + tags: [upstream_flank] + sequence: gtgatgtcaaagcttgaaaaaacgcacgtaacaaaagcaaaatttatgctccatgggggagactacaaccccgatcagtggctggatcggcccgatattttagctgacgatatcaaactgatgaagctttctcatacgaatacgttttctgtcggcatttttgcatggagcgcacttgagccggaggagggcgtatatcaatttgaatggctggatgatatttttgagcggattcacagtataggcggccgggtcatattagcaacgccgagcggagcccgtccggcctggctgtcgcaaacctatccggaagttttgcgcgtcaatgcctcccgcgtcaaacagctgcacggcggaaggcacaaccactgcctcacatctaaagtctaccgagaaaaaacacggcacatcaaccgcttattagcagaacgatacggacatcacccggcgctgttaatgtggcacatttcaaacgaatacgggggagattgccactgtga +DS_lacA: + description: downstream flank lacA + prefix: GTCT + suffix: AAGC + tags: [downstream_flank] + sequence: ttcaagctatatttggagttgagcctcttgaaacggacaccctgtatccgaaggatcgaaacgctgtcagctaccgcagccaaatatatgaaatgaaggattatgcaaccgtgattgatgtaaagacagcttcagtggaagcggtgtatcaagaagatttttatgcgcgcacgccagcggtcacaagccatgagtatcagcagggcaaggcgtattttatcggcgcgcgtttggaggatcaatttcagcgtgatttctatgagggtctgatcacagacctgtctctctctccagtttttccggttcggcacggaaaaggcgtctccgtacaagcgaggcaggatcaggacaatgattatatttttgtcatgaatttcacggaagaaaaacagctggtcacgtttgatcagagtgtgaaggacataatgacaggagacatattgtcaggcgacctgacgatggaaaagtatgaagtgagaattgtcgtaaacacacattag +eryR(Bsub): + description: erythromycin resistance in Bacillus subtilis + prefix: AAGC + suffix: ATAG + tags: [selection] + sequence: GAGTTAACAAGAGTTTGTAGAAACGCAAAAAGGCCATCCGTCAGGATGGCCTTCTGCTTAGCTAGAGCGGCGGATTTGTCCTACTCAGGAGAGCGTTCACCGACAAACAACAGATAAAACGAAAGGCCCAGTCTTTCGACTGAGCCTTTCGTTTTATTTGATGCCTCAAGCTAGAGAGTCCTAGAGTCTAGGGACCTCTTTAGCTCCTTGGAAGCTGTCAGTAGTATACCTAATAATTTATCTACATTCCCTTTAGTAACGTGTAACTTTCCAAATTTACAAAAGCGACTCATAGAATTATTTCCTCCCGTTAAATAATAGATAACTATTAAAAATAGACAATACTTGCTCATAAGTAACGGTACTTAAATTGTTTACTTTGGCGTGTTTCATTGCTTGATGAAACTGATTTTTAGTAAACAGTTGACGATATTCTCGATTGACCCATTTTGAAACAAAGTACGTATATAGCTTCCAATATTTATCTGGAACATCTGTGGTATGGCGGGTAAGTTTTATTAAGACACTGTTTACTTTTGGTTTAGGATGAAAGCATTCCGCTGGCAGCTTAAGCAATTGCTGAATCGAGACTTGAGTGTGCAAGAGCAACCCTAGTGTTCGGTGAATATCCAAGGTACGCTTGTAGAATCCTTCTTCAACAATCAGATAGATGTCAGACGCATGGCTTTCAAAAACCACTTTTTTAATAATTTGTGTGCTTAAATGGTAAGGAATACTCCCAACAATTTTATACCTCTGTTTGTTAGGGAATTGAAACTGTAGAATATCTTGGTGAATTAAAGTGACACGAGTATTCAGTTTTAATTTTTCTGACGATAAGTTGAATAGATGACTGTCTAATTCAATAGACGTTACCTGTTTACTTATTTTAGCCAGTTTCGTCGTTAAATGCCCTTTACCTGTTCCAATTTCGTAAACGGTATCGGTTTCTTTTAAATTCAATTGTTTTATTATTTGGTTGAGTACTTTTTCACTCGTTAAAAAGTTTTGAGAATATTTTATATTTTTGTTCATGTAATCACTCCTTCTTAATTACAAATTTTTAGCATCTAATTTAACTTCAATTCCTATTATACAAAATTTTAAGATACTGCACTATCAACACACTCTTAAGTTTGCTTCTAAGTCTTATTTCCATAACTTCTTTTACGTTTCCGCCATTCTTTGCTGTTTCGATTTTTATGATATGGTGCAAGTCAGCACGAACACGAACCGTCTTATCTCCCATTATATCTTTTTTTGCACTGATTGGTGTATCATTTCGTTTTTCTTTTGTGCTAGAGGATCAATTCTTGAAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTG +US_thrC: + description: upstream flank thrC + prefix: ATTA + suffix: CGAG + tags: [upstream_flank] + sequence: aattcatgtaaaagatgaggttggttcattctcgaaaattacatctgtgttctcagagcggggcgtgagctttgaaaaaatccttcagctgccaattaaaggccatgatgagttagctgaaatcgtaattgtcacacatcatacatcagaagctgatttcagtgatatcctgcaaaacctaaatgatttggaagtcgttcaagaagtcaaaagcacatatcgtgtagaagggaacggttggagctaatgtggaaaggacttatccatcaatataaagaatttttacctgtaacagatcaaacaccggcgctaactttacatgaaggaaacacacctcttattcacctgccgaagctgtctgagcagctcggaattgagcttcatgtcaaaacggaaggcgtcaatcctacgggatcatttaaagatcgcggaatggttatggctgtggcaaaggcaaaagaagaaggcaatgacacgattatgtgcgcgtcaacaggtaacacttccgctgctgcggcagcatatgcagcccgtgctaacatgaaatgcattgtcatcatcccgaacggaaaaattgcatttggaaaactcgctcaagctgtcatgtacggagccgagattatcgcaattgacggaaactttgacgatgcgcttaaaattgtccgttccatctgtgagaaatcaccgattgcccttgtcaactcagtcaacccttaccgcattgaaggccaaaaaactgctgccttcg +DS_thrC: + description: + prefix: GTCT + suffix: AAGC + tags: [downstream_flank] + sequence: AGGCCTTCCGAAAATGCGCGGCTTTGAAGCTGAGGGTGCGGCGGCAATCGTGCGCAATGAAGTGATTGAAAATCCGGAAACAATAGCGACAGCCATTCGTATCGGAAACCCGGCAAGCTGGGACAAAGCTGTAAAGGCAGCCGAGGAATCCAATGGGAAAATTGACGAAGTCACTGATGATGAAATCCTTCACGCATATCAGCTGATCGCCCGTGTAGAAGGCGTGTTTGCAGAACCAGGTTCTTGCGCGTCTATCGCAGGAGTGCTGAAACAGGTGAAATCCGGAGAAATTCCGAAAGGCAGCAAGGTCGTAGCTGTGTTAACAGGAAACGGACTGAAAGATCCGAACACAGCGGTCGACATTTCAGAAATCAAGCCTGTCACATTGCCGACTGATGAAGACAGCATCCTTGAATATGTAAAAGGAGCGGCCCGTGTATGAACGAAGCCGACATGCTGTTCTCTGTCACTGTTCCCGGAAGCACAGCTAACCTAGGCCCCGGCTTTGATTCAGTCGGAATGGCGCTCAGCAGATATTTGAAGCTGACCGTCTTTGAAAGCGACAAATGGTCTTTTGAGGCTGAAACAGAAACAGTCGCCGGAATT +Pars: + description: Promoter of ars operon. Repressed by ArsR, induced by arsenate or arsenite. + prefix: TACA + suffix: AACT + tags: [promoter] + sequence: ttaatcaaaataaattgatttatttgcttgcattaatttaaaaatcatgagtataataaatacatcaaaa +P43: + description: Strong promoter activity during exponential and stationary phase. + prefix: TACA + suffix: AACT + tags: [promoter] + sequence: tattttacatttttagaaatgggcgtgaaaaaaagcgcgcgattatgtaaaatataa +PxylA_xylR: + description: Full xylose inducible promoter + prefix: TACA + suffix: AACT + tags: [promoter] + sequence: AGCGGATAACAATTTCACACAGGAGGCCGATCGCGTTATGCAGCAATGGCAAGAACGTCCCGGGGAGCTCCTAACTTATAGGGGTAACACTTAAAAAAGAATCAATAACGATAGAAACCGCTCCTAAAGCAGGTGCATTTTTTCCTAACGAAGAAGGCAATAGTTCACATTTATTGTCTAAATGAGAATGGACTCTAGAAGAAACTTCGTTTTTAATCGTATTTAAAACAATGGGATGAGATTCAATTATATGATTTCTCAAGATAACAGCTTCTATATCAAATGTATTAAGGATATTGGTTAATCCAATTCCGATATAAAAGCCAAAGTTTTGAAGTGCATTTAACATTTCTACATCATTTTTATTTGCGCGTTCCACAATCTCTTTTCGAGAAATATTCTTTTCTTCTTTAGAGAGCGAAGCCAGTAACGCTTTTTCAGAAGCATATAATTCCCAACAGCCTCGATTTCCACAGCTGCATTTGGGTCCATTAAAATCTATCGTCATATGACCCATTTCCCCAGAAAAACCCTGAACACCTTTATACAATTCGTTGTTAATAACAAGTCCAGTTCCAATTCCGATATTAATACTGATGTAAACGATGTTTTCATAGTTTTTTGTCATACCAAATACTTTTTCACCGTATGCTCCTGCATTAGCTTCATTTTCAACAAAAACCGGAACATTAAACTCACTCTCAATTAAAAACTGCAAATCTTTGATATTCCAATTTAAGTTAGGCATGAAAATAATTTGCTGATGACGATCTACAAGGCCTGGAACACAAATTCCTATTCCGACTAGACCATAAGGGGACTCAGGCATATGGGTTACAAAACCATGAATAAGTGCAAATAAAATCTCTTTTACTTCACTAGCGGAAGAACTAGACAAGTCAGAAGTCTTCTCGAGAATAATATTTCCTTCTAAGTCGGTTAGAATTCCGTTAAGATAGTCGACTCCTATATCAATACCAATCGAGTAGCCTGCATTCTTATTAAAAACAAGCATTACAGGTCTTCTGCCGCCTCTAGATTGCCCTGCCCCAATTTCAAAAATAAAATCTTTTTCAAGCAGTGTATTTACTTGAGAGGAGACAGTAGACTTGTTTAATCCTGTAATCTCAGAGAGAGTTGCCCTGGAGACAGGGGAGTTCTTCAAAATTTCATCTAATATTAATTTTTGATTCATTTTTTTTACTAAAGCTTGATCTGCAATTTGAATAATAACCACTCCTTTGTTTATCCACCGAACTAAGTTGGTGTTTTTTGAAGCTTGAATTAGATATTTAAAAGTATCATATCTAATATTATAACTAAATTTTCTAAAAAAAACATTGAAATAAACATTTATTTTGTATATGATGAGATAAAGTTAGTTTATTGGATAAACAAACTAACTCAATTAAGATAGTTGATGGATAAACTTGTTCACTTAAATCAAAGGCTGAG +mRuby_with_RBS(Bsub): + description: mRuby with ribosomal binding site for Bacillus + prefix: AACT + suffix: ATCC + tags: [rbs_cds, reporter] + sequence: +US_glmS: + description: upstream glmS flank + prefix: ATTA + suffix: CGAG + tags: [upstream_flank] + sequence: CAACATGCCGCTTCTGTCTAAGAAACCGCTCTTCATTTTCCTTTCTCAAAGCGGAGAAACAGCAGACAGCCGCGCGGTACTCGTTCAAGTCAAAGCGCTCGGACACAAAGCCCTGACAATCACAAACGTACCTGGATCAACGCTTTCTCGTGAAGCTGACTATACATTGCTGCTTCATGCAGGCCCTGAGATCGCTGTTGCGTCAACGAAAGCATACACTGCACAAATCGCAGTTCTGGCGGTTCTTGCTTCTGTGGCTGCTGACAAAAATGGCATCAATATCGGATTTGACCTCGTCAAAGAACTCGGTATCGCTGCAAACGCAATGGAAGCTCTATGCGACCAGAAAGACGAAATGGAAATGATCGCTCGTGAATACCTGACTGTATCCAGAAATGCTTTCTTCATCGGACGCGGCCTTGACTACTTCGTATGTGTCGAAGGCGCACTGAAGCTGAAAGAGATTTCTTACATCCAGGCAGAAGGTTTTGCCGGCGGTGAGCTAAAGCACGGAACGATTGCCTTGATCGAACAAGGAACACCAGTATTCGCACTGGCAACTCAAGAGCATGTAAACCTAAGCATCCGCGGAAACGTCAAAGAAGTTGCTGCTCGCGGAGCAAACACATGCATCATCTCACTGAAAGGCCTAGACGATGCGGATGACAGATTCGTATTGCCGGAAGTAAACCCAGCGCTTGCTCCGTTGGTATCTGTTGTTCCATTGCAGCTGATCGCTTACTATGCTGCACTGCATCGCGGCTGTGATGTGGATAAACCTCGTAACCTTGCGAAGAGTGTTACTGTGGAGTAATAAATGTTTAACCCCTTTGGATAAGATTATCTAAAGGGGTGT +DS_glmS: + description: downstream glmS flank + prefix: GTCT + suffix: AAGC + tags: [downstream_flank] + sequence: TTATGTCCAAAAGCGAAAGAACGTCTACAGTGGAAAAATGGATTAAAGAAGTTGATGGTACAGGACCCGACTATCGATCGACCATGGCTATAGATTTAAGATGTTTCCTCATTAGATCGGTCAATCAGGTTAAAAGGCATTAAAACTTGGAGAGAAACTGCTTTTGCCTAACTGAGTTTTCTAATGTTAATACAGATATAAGAGAGCAGTTCCTCATTAAGATACGCTTTTGATAGCAGTTGGAATTTAGCATGCTAGAAATCCGAAAACCGGAGAGCCTATTGGAATGATAACGTGTTTTTTTATTGACAATTAATAAAGGGCAAAGAATTCTAGAAGTCGCACTTACAATGTACTTGTGGGATTAGGTTTTAAAGTTCATTCTTTTTTTATTGAGTACATTCCGGTTAAAGTTGAAATGATTCAGTAAAGGACACGCCATAAATAAAAGGTAGCGTATGATTGCCATTCTTTCCATGGAACCGATATTTCTAAGATTTCATCTTTCGTAGGCTTTCGGTTCATATTACGTAATATTTTTATCGAATGAATAAGACCCACATCATCGATTGGAAAAGCTGTCGGGAACCTAAGGCAGCGCATTAGAACATAATTGGCTGTCCAAGGACCGATTCCTCGTATTTTAATTAAGTTTTTTTCAGCATCTTTAAAGTTCATTTTCATTAATTTTTCCCTCGATAATTCTCCACTTGCCATTAATCTGGCTATTCCAATGATATATTCACTTTTTTTCACCGTCATTTTAATATCTGCCAGGTCGGTAGGGGTTAACCGTGCAATTCGCTCGTACGGTGGGAACACCCAATACTTTTTACCATTCCATTCAATAGAATCGCCAAATGCTTCTACAAATTGCTTCTTTAAGGAGTACGCGAAGGCTAA +Pmtl-short: + description: Pmtl promoter. Mannitol inducible + prefix: TACA + suffix: AACT + tags: [promoter] + sequence: tcctgaaaccaggagcctttttatttttaaaaaattgtcacagtcatgtgccaaagtcctctttactttcaattgtatagggactgtaagcgttttaacatagagtcaaagggaagcatcattctctgaacatattt +Ptet_tetR: + description: Ptet inducible promoter with tetR. Tet inducible + prefix: TACA + suffix: AACT + tags: [promoter] + sequence: TTAAGACCCACTTTCACATTTAAGTTGTTTTTCTAATCCGCATATGATCAATTCAAGGCCGAATAAGAAGGCTGGCTCTGCACCTTGGTGATCAAATAATTCGATAGCTTGTCGTAATAATGGCGGCATACTATCAGTAGTAGGTGTTTCCCTTTCTTCTTTAGCGACTTGATGCTCTTGATCTTCCAATACGCAACCTAAAGTAAAATGCCCCACAGCGCTGAGTGCATATAATGCATTCTCTAGTGAAAAACCTTGTTGGCATAAAAAGGCTAATTGATTTTCGAGAGTTTCATACTGTTTTTCTGTAGGCCGTGTACCTAAATGTACTTTTGCTCCATCGCGATGACTTAGTAAAGCACATCTAAAACTTTTAGCGTTATTACGTAAAAAATCTTGCCAGCTTTCCCCTTCTAAAGGGCAAAAGTGAGTATGGTGCCTATCTAACATCTCAATGGCTAAGGCGTCGAGCAAAGCCCGCTTATTTTTTACATGCCAATACAATGTAGGCTGCTCTACACCTAGCTTCTGGGCGAGTTTACGGGTTGTTAAACCTTCGATTCCGACCTCATTAAGCAGCTCTAATGCGCTGTTAATCACTTTACTTTTATCTAATCTAGACATCATTAATTCCTCCTTTTTGTTGACATTATATCATTGATAGAGTTATTTGTCAAACTAGATTTTTATTTCGATGCCCTGGACTTCATGAAAAACTAAAAAAAATATTGACACTCTATCATTGATAGAGTATAATTAAAATAAGCTTGATCGTAGCGTTAACAGATCTGAGCTCCTGCAGTAAGCTGAG +Pman_manR: + description: Pman inducible promoter with manR. Inducible with mannose. + prefix: TACA + suffix: AACT + tags: [promoter] + sequence: ttaaagtgtgaataataagatcttgtctcaactgtataccgaaatcagctcattaaaatcgctttttttaccataggttccggtaataaaggcatttttccctataacaaaaaaagcaaggaataatccctgcttttaataatccaaatgagataaaaatgtcatgacattggtttctttcgtcagcttctggacgaaggccggctgctcgctaagatatgaaagctcgctaaacagctgctttgtcatagtttgatcctcgtgtttgacagccagcataaaaacgagcgaaactttttcatttccccattcaagcggctcctttaatgtggcaatcgcgattgctgattgtttaataaatttagcgtttgcatgaggaatggcaatgcctgacccaatatttgtagccgacattttttccctcatcaccgcatgaaccgcatagtccttatcaacatagcctttctcaaacaaagctgtcgccagctgttcgattaacttgtagcgatgctccgcctcctgctgcaaaaaaaccaaaaaaggagtcgtattgttcagcatctgaaacgttttttgtttttggcggtgtgattcgcccagctgacgtataaacgcacttaatttcttctcttcgccaggctctaaaagcggagacacaacaatatggggaaccgttatattctccagcgcaatggtggatatgacaagatcaatatcctcatgctttttgatataatcctttaaatcggcctttgcaatacaagccattacagcgatttggtgatatttccgttcaatttttgtccgcagcaattgtgacatcccaatccccatgtggcagacgataaccgcctttttatgtgtctcgctgctgccctgcatccgttcgatcgctgcctgaaaatgcaatgtcaaatatgctgcctcttcctcaggaatatgaagatcaaaagtttgattgatgtcttccaaaacatcaatgatgaggtggaacaaataagggtacatctttttgatgtcattgagcataggatttgctacagaaagatcgtagctcagccgttgtagcacagtattgagatgaatgttcaaaccattgatcaaatcctgatccttgtgaaaatccatcattttcagctctgaaacacgattgattaaatgccctacaaccttaggaagcacagcattttcaaggttctcttctgtctgcaatggataacgaaccttaccgcccaatatatgtaatgttaagtacacggcttcttcttcagggaagcgaattgcaaaaaccggttcaagccgctgtaaacaagcaaatgtccattgatattctttcttcttttttactgccgccatttcttttggggacaatgaaatcggctgcttcattttgatgcggcggaccattaacaatgtatgcagcaacaaactttcaaacgtttcgtctgtgaaatagagggaatgctttttttgtagtgatttgatttctttcgtcacaaagtctacttcataatgtaaaaacttacttttaataaattggctggtaaacgctgtattgtgaatcaaatcagatattcttgccaatgcctttcttttatttctttcgttcccttcaacctttaatccaagccgctgcctagaaaccagtgtgagatcaaaacgcttcagccactcttctactgcgtataaatcctttttaatggaagatcgattgacaaaatgccgagcggcaatgtcttttgcggaaacaggcttcggattcattaataagtcataagcaatgtgcagcattctttccttatcgctccgctgcctagaagaaaaatgttcagtatgtaattgttggcttagccaggtcctttcctgttcttctatatgtaaataaacgcccaaccctggttttcgaataagctgtgcatgagaatgctcattgagataatcctcgatcactttcaaatcattccggattgttttttcagaacactgtaccctatctgcgaaatcttgcacaactaaatagtcatcaggttcagataacaacagatacaaaatctctttttgtcttgtatttatgtattccatctgtttttctatccttcctgtcagtatctacttgagcaaattgatagtataagtatactatataagaaaccgtttacattataaaacaaaatcatagagacaaaagtttcgttttttaccgaagcttccggaaaaaagcgatttttatactctttgagtttgctatg +P3P: + description: Ultra strong constitutive promoter. 42-13x stronger than Pveg + prefix: TACA + suffix: AACT + tags: [promoter] + sequence: tcttttggaagaaaatatagggaaaatggtatttgttaaaaattctgaatatttatacaatatcatatgtttcacattgaaatatgctgtccagactgtccgctgtgtaaaaaataggaataaaggggggttgttattattttactgatatgtaaaatataatttgtataagaaaattcgaaacgtaagatgaaaccttagataaaagtgctttttttgttgcaattgaagaattattaatgttaagcttaattaaagataatatctttgaattgtaacgcccctcaaaagtaagaactacaaaaaaagaatacgttatatagaaatatgtttgaaccttcttcagattacaaatatattcggacggactctacctcaaatgcttatctaactatagaatgacatacaagcacaaccttgaaaatttgaaaatataactaccaatgaacttgttcatgtgaattatcgctgtatttaattttctcaattcaatatataatatgccaatacattgttacaagtagaaattaagacacccttgatagccttactatacctaacatgatgtagtattaaatgaatatgtaaatatatttatgataagaagcgacttatttataatcattacatatttttctattggaatgattaagattccaatagaatagtgtataaattatttatcttgaaaggagggatgcctaaaaacgaagaacattaaaaacatatatttgcaccgtctaatggatttatgaaaaatcattttatcagtttgaaaattatgtattat +US_sigF: + description: upstream flank sigF + prefix: ATTA + suffix: CGAG + tags: [upstream_flank] + sequence: gctgaacttggaggacctttcctttatggacagctcggggcttggcgttattttaggaagatacaagcaaattaagcaaattggcggagaaatggttgtttgcgctatctctcctgcggtgaagcgattgtttgatatgtcgggtctgtttaaaattatccgatttgaacaatctgaacagcaggcactcctgacactgggggtggcatcatgaaaaatgaaatgcaccttgagttttctgccctcagtcagaatgaatcgttcgcccgtgtgacagttgcttcatttatagctcagctggacccgacaatggatgaactgactgaaatcaaaacagtcgtgtcagaggctgtcacgaatgcgattatccatggatatgaagagaactgtgaagggaaagtttacatttcagtgacgctggaagatcatgtcgtatatatgactattcgtgatgaaggcttaggcattacagatcttgaagaagcccgtcagcctctatttacgactaagcctgagcttgagcgctctggaatgggctttaccattatggaaaatttcatggatgatgtcagtatcgattcatcgcctgaaatgggaacaacgattcgcttaacaaagcacttatcaaaaagcaaagcgctttgtaattaaggagatttgttatggatta +DS_sigF: + description: downstream flank sigF + prefix: GTCT + suffix: AAGC + tags: [downstream_flank] + sequence: tctgcagtgcaggctagcttttttgtgcaaaagcgtggtaatttatggtcttttcgagcggatgaatgagaacaaaatcgaaccacatactacatatataaccaccgaaagatggtgatcaatgatggaacgacgaatatttatccggcttcgccaccgagtgctggcacatccaggggatattattaccgttggagatgccgcgcaaatagaagggcagcttcagctgaaaaagaaactttcggctatgccgctttatcaggtgagcgaaaaagataaaaatatcgtaattctggatatcatacaagtcctcagagccattcatttacaagacccgacaattgatgttcaaaccgtaggcggagcagaaaccattgttgaaattcagtatcgaaagcgaaatttatcaacggttctatttatcggtgtctggctgcttctgtttattggatcgtgtcttgccatcatgaactttcatgaggatgtaagcatgagagatgttcatatcgcactatatgaaatcataaccggagagaggaatgactatccatatttgcttcaaatcccatacagcatcggtttgggactggggatgatcgtgttttttaaccacatatttaaaaagcgcctaaatgaagagcccagcccgctggaggttgagatgtttaactatcagcttgatctcgatcaatatgtggccatgcatgagaatcaagaaaccataaaggatctgcatgatcgttagtgtattgttcatcatttttgtcgggctcggcggaggcataacggtgggggcgggtttcgttgcttttttaaccgtaatgggaatcattccgcggctgatgcagctcaccaaaacaatgagatttgttcaggcttatgaagcg +kanR(Bsub): + description: bacillus subtilis kanamycin resistance + prefix: AAGC + suffix: ATAG + tags: [selection] + sequence: gaggtcatcgttcaaaatggtatgcgttttgacacatccactatatatccgtgtcgttctgtccactcctgaatcccattccagaaattctctagcgattccagaagtttctcagagtcggaaagttgaccagacattacgaactggcacagatggtcataacctgaaggaagatctgattgcttaactgcttcagttaagaccgaagcgctcgtcgtataacagatgcgatgatgcagaccaatcaacatggcacctgccattgctacctgcacagtcaaggatggtagaaatgttgtcggtccttgcacacgaatattacgccatttgcctgcatattcaaacagctcttctacgataagggcacaaatcgcatcgtggaacgtttgggcttctaccgatttagcagtttgatacactttctctaagtatccacctgaatcataaatcggcaaaatagagaaaaattgaccatgtgtaagcggccaatctgattccacctgagatgcataatctagtagaatctcttcgctatcaaaattcacttccaccttccactcaccggttgtccattcatggctgaactctgcttcctctgttgacatgacacacatcatctcaatatccgaatagggcccatcagtctgacgaccaagagagccataaacaccaatagccttaacatcatccccatatttatccaatattcgttccttaatttcatgaacaatcttcattctttcttctctagtcattattattggtccattcactattctcattcccctttcagataattttagatttgcttttctaaataagaatatttggagagcaccgttcttattcagctattaaacccattatatcgggtttttgaggggatttcaactgcag +US_yhgE: + description: upstream flank yhgE + prefix: ATTA + suffix: CGAG + tags: [upstream_flank] + sequence: gacgggtctaaacaggtccaaagcggatcatcaaaacttgcagacggatctgcatcacttgatacaggtttaggcaagctattggacggcacaggagagctgtctagtaagctgaaggatgcggctgaccaaacaggagatatcgatgctgatgatcaaacatacggcatgtttgcggatccggttaaaaccaaagatgatgccattcattccgttcctaactatggaacagggctgacaccttatatcctgtcaatgggcttgtatgtcggcggcatcatgctgacggttgtcttcccgctgaaggaagcgtcaggacgtccgagaaacggttttgagtggttcttcagcaaattcaatgtcatgatgcttgtaggaatcattcaatcattgattgtggcgacggttcttctattgggaattggtcttgaggtagagagcacgtggagattttacgtgtttacgatcattacgagtctcgctttcctggcgataattcaatttttggctacaacgatgggcaacccggggcgatttatcgcagtcattattttagtgctgcagctgggagcgagcggaggaaccttcccgctggaattgcttccgaatttctatcaggtcatccacggcgcattgccgatgacctacagcattaacggattcagagcagtcatttcaaacggtgactttggctacatgtggcagatggccggcgtcttaatcggcattgccctcgtcatgattgcactaagcattacttactttacaatgttaagccggaaggaagaaacatcagaagaacagccggcttcataa +DS_yhgE: + description: downstream_flank yhgE + prefix: GTCT + suffix: AAGC + tags: [downstream_flank] + sequence: agaaaaagaacttgtttccttggaaataagttctttttttgttacatcccccatttaataagcaatcctgtataggttaatccgccgccaaacccgaaaagcaaaacgatttgatcttttttcagcttcccggctttcactgcgagatcgagcgccaaaacaattgaaaccgaagacgtgtttccgtagtgctcaacactagtgagcgttttttcaatcgggaacggtgttttttcacaaattgactcgatcatgcgcaagttggcgctgtgaggaacaaaccaatcgagatcatcggagctgagtcctgctttatgtaaaagccgttcaaattcgccagggacggttcttgcggcccatttatatacctcgcgtccgttttgcaccatttttccggaaccgacaagctgcacaccgtttatttcatttcgcagtccggcacgatacaaaatatcgccgccgttcccgcttgttccttgtacagacgcaagaaatcccggcgtctcttcatctcgttctactaacagcgcacccgcggcatcgccgaacagtacgcatgtcgttcgatcggtataatcggttacctttgataacgtctctccggcgatgacgagaattttttgatgaaggccagatgtgatcaatccatttgccaaatggaggccgtatgtcagcccggcgcatgtcgcattaatatccagcgcgccggtgctttcccagccgaaatattcctgtacgcggcatgccgtactcggaaaggcgtaatcggatgtggttgtg +spcR(Bsub): + description: bacillus subtilis spectinomycin resistance + prefix: AAGC + suffix: ATAG + tags: [selection] + sequence: acgtgtaggctggagctgcttccctgcaggagtcaggtcaatcatcagataactatggcggcacgtgcattaaccacggttgtatcccgtctaaagtactcgtctgtgcaacaggtgacatcacgaaacgatcctcatcctgtctcttgatccatggattacgcgttaacccgggcccgcggatgcatatgatcagatcttaaggcctaggtctagaggatcgatctgtataataaagaataattattaatctgtagacaaattgtgaaaggatgtacttaaacgctaacggtcagctttattgaacagtaatttaagtatatgtccaatctagggtaagtaaattgagtatcaatataaactttatatgaacataatcaacgaggtgaaatcatgagcaatttgattaacggaaaaataccaaatcaagcgattcaaacattaaaaatcgtaaaagatttatttggaagttcaatagttggagtatatctatttggttcagcagtaaatggtggtttacgcattaacagcgatgtagatgttctagtcgtcgtgaatcatagtttacctcaattaactcgaaaaaaactaacagaaagactaatgactatatcaggaaagattggaaatacggattctgttagaccacttgaagttacggttataaataggagtgaagttgtcccttggcaatatcctccaaaaagagaatttatatacggtgagtggctcaggggtgaatttgagaatggacaaattcaggaaccaagctatgatcctgatttggctattgttttagcacaagcaagaaagaatagtatttctctatttggtcctgattcttcaagtatacttgtctccgtacctttgacagatattcgaagagcaattaaggattctttgccagaactaattgaggggataaaaggtgatgagcgtaatgtaattttaaccctagctcgaatgtggcaaacagtgactactggtgaaattacctcgaaagatgtcgctgcagaatgggctatacctcttttacctaaagagcatgtaactttactggatatagctagaaaaggctatcggggagagtgtgatgataagtgggaaggactatattcaaaggtgaaagcactcgttaagtatatgaaaaattctatagaaacttctctcaattaggctaattttattgcaataacaggtgcttacttttctggagttctttagcaaatttttttattagctgaacttagtattagtggccatactcctccaatccaaagctatttagaaagattactatatcctcaaacaggcggtaacgcgttgtccttttccgctttaataaactatggaagtatgtacagtcttgcaatgttgagtgaacaaacttccataataaaatatgcaagctggtaccctaaggaggatattcatatgcaagcact +US_yrbD: + description: upstream flank yrbD + prefix: ATTA + suffix: CGAG + tags: [upstream_flank] + sequence: gtctgaattgcctgctgtgatctcattaatattcaaaagcgccttcgcattggattccgcttttggcggcttgatcggtatggcgatttcctggggcgttaaacgtggcatttactcaaacgaagccggccagggaacaggcccgcatccggcagctgctgcagaggtttctcatccggtcaagcagggactggttcaggcattttctgtttatattgacaccttgtttgtatgttccgctacagctttcatgatcttgtttaccggcatgtacaatacccaggctgcagatggttcctttatcgttcaccagcttaaaggcgtagaagcaggacctggtttcacacaagcggcaattgacagtgttcttcccggattcggtgcaggctttgttgccatcgctttattcttcttcgcttttacaacaattatggcgtattactacatcgcagaaacaaatatcgcctatttagcacgaggcagagaaagcaaatgggcgatgcttggcttaaaactgattattttagccgccacgttctatggtactgtgaaaacagcttcacttgcttgggcattaggggatgcagggcttggcattatggtatggctgaacgtcattgccattgtgctgcttgccaagccggcgctccttgctttaaaggattatgagcgccaaaagaagcagggcttagacccgatctttgatccaaaagcgctgggcatcaaaaacgctgatttctgggagaaagaatacactcatgaaagtgaacgtgtctcctgaacatactaaaaccggcccgatatgacctcgtgccggtttt +DS_yrbD: + description: downstream flank yrbD + prefix: GTCT + suffix: AAGC + tags: [downstream_flank] + sequence: ttatgaacgatagtttgtatacacttcttgaacatcatcgtcatcttcaaggacgtctattaatgtttccagcttctcaacagcctgatcatcggcttctgcataggtattgggaagcatggtcacctccgccgaagaaattagatatttcgattcgagggctgttttcacctcttcaaaatgctcaggctctgtgaaaacctcatataggtcttcttcaattcgcagctcttctcctcctgcttcaagcacatcaagcatgagctcatcttcttctatttgccgttcagtacggtctatcgtaatgaacccctttcgttcaaataaaaaggctacacatccgctttctcccagacttccgccatttttattaaaggctgtccgcacatttgacgccgttcggtttttattatctgttacgcattcaaccatgacggcaatccctgaagggccgtagccttcataggtgatttcttcatagctgcttccgtcttggcctcccgcagcttttttaatggcccgatctatattttcattcggcatattggcgccttttgctttttcgatgacaagacgcaatgctgaatttgcttctggatccgctccgccttctttcgctgccacatagatttcttttgccagcttcataaatattttcccgcgctttgcatcctgtgcattttttcttttttgaatgtttttccacttggaatggcctgccatgttttcaccttcttttacattttctatgtacactttaatgggaaatgttcgctgatgcaaagaaaaaggctgc +TO_pE194ts: + description: pE194ts is a temperature sensitive origin in Bacillus subtilis. Can be used for curable plasmids. + prefix: GTCT + suffix: AAGC + tags: [origin] + sequence: gacacctaaattcaaaatctatcggtcagatttataccgatttgattttatatattcttgaataacatacgccgagttatcacataaaagcgggaaccaatcatcaaatttaaacttcattgcataatccattaaactcttaaattctacgattccttgttcatcaataaactcaatcatttctttaattaatttatatctatctgttgttgttttctttaataattcatcaacatctacaccgccataaactatcatatcttctttttgatatttaaatttattaggatcgtccatgtgaagcatatatctcacaagacctttcacacttcctgcaatctgcggaatagtcgcattcaattcttctgtaattatttttatctgttcataagatttattaccctcatacatcactagaatatgataatgctcttttttcatcctatcttctgtatcagtatccctatcatgtaatggagacactacaaattgaatgtgtaactcttttaaatactctaaccactcggcttttgctgattctggatataaaacaaatgtccaattacgtcctcttgaatttttcttgttttcagtttcttttattacattttcgctcatgatataataacggtgctaatacatttaacaaaatttagtcatagataggcagcatgccagtgctgtctatctttttttgtttaaaatgcaccgtattcctcctttgcatatttttttattagaataccggttgcatctgatttgctaatattatatttttctttgattctatttaatatctcattttcttctgttgtaagtcttaaagtaacagcaacttttttctcttcttttctatctacaaccatcactgtacctcccaacatctgtttttttcactttaacataaaaaacaaccttttaacattaaaaacccaatatttatttatttgtttggacaatggacaatggacacctaggggggaggtcgtagtacccccctatgttttctcccctaaataaccccaaaaatctaagaaaaaaagacctcaaaaaggtctttaattaacatctcaaatttcgcatttattccaatttcctttttgcgtgtgatgcg +TO_pHT315: + description: pHT315 ~15 copies in Bacillus subtiis. NOTE 897 was mutated G->A to prevent BsaI cut site. Theta replication. + prefix: GTCT + suffix: AAGC + tags: [origin] + sequence: ccatcctccaaagttggagagtgagttttatgtcgcaaatattaatgtttctggtgaaccttatcaaattttcgttgatttaatagaaacatagcggtaaaattagcagtaacttaatagaacggaaatgaaaaaagccactctcatatgctattggctaccaacctttagcgagaatgacttaatcctgtacagccatacaggacttcgacttataagaggcgccaacttcaaataagttatttgccttgttttcgcgaacaaggcttattagatacacctattgtaccgttactctacgaatatttcagctagtaattactagcattgtcatatacataataaaacggatataaaagggcgttttctatacctagaagtcttgtaaatgtacagggcgtttagatatagagaacgccctttttgtgttccgttccagtggaagctaccactttaaaaagatggtctagtgtagccaatgcaggagagtacactcggatatcagttgtcgttgcattcaactgtctgacgtaagcgaggtaaaggacacaagccttgcataaaacaagcctacgggatgtaaatcctaataatgatgataaccaagacgttagcggcaaaaagtgttgggggttcaaaataagacatgattgtgcgactggagttaaacagttactcgtaagcggcgatcatgacactgattcacggctattcttgtacaagctagctttattacaaggatatgcgggttatatagcgaatcacccgaaagggaacggtgttgggcgtgagaaacgcaccgtacggcgcaatacaatgccaataagctatatacggacggtatagtagttttgtaagctataaccgtttgtcgtcaatgcaaccaatctcaattcAagacctcggcatctaagccagtacgaatgagtgggcgttttaacctcgtaaattttcaacaggggttactatgcccaaaactacattcagatttcctaacaaactcgccagtatgaaaaccttaagaccttaaagtcaagggatttgaaggattttaacctcgattagcaaaaaatgtagagtactgaagcaactaccattaactaagatagtgggggattgaggaagaatccagagctgtttaaatcaagtgaaagacaagatgaaattaaaagaatagtgaaagataggggagtggttctctatgagaaaggaaatggctagagaacaaaggcagcggtttattgatctattgttagactttatggtaaagaatcctcatttatttgttaatggtacagaggatgaaagtaataatgttgttacaaaatgtaatagtgatattaaagaggttgcggagtcatatttaactcttttatagtgagagggttaaaactaattaatatgtattaaggcccaatgttggaattattgtatttcactaggcaacctacttactaaaagtaagattatccattagtggatgttataatattgggttttttaacacaataatcatcgcctttcggtgtcgtttgatagaaaagtaaccattagcgatgaaaaagtcaatataaaaagccatccgtaaaaaacggatggcttaccgtacataggatcgttggtagggcggcgtatcctacatctctggtaacttacctagccaatcaaatgcttgagaacggcggttagataagcgcgtggggaacctttcccacctcaaagatcctatatcattattatgttactttctacaggtagtataccatgttcttatattttagtaaactccccgttagcttaacaggtctttgtaagcaattaaacgtccactattcaatcgtctttggattttcgcaggaccgttttttagatcgaacatagttgataagaacaaataaccgcttgggtccaactttatagcaattagtatatggtcatttaaaatctttaccaattcaacgctattaggttctttaggattttgcccgacatagtcggggtgttcaacgatatcttttatgtgcgatgaatatttttcataaataccaggatgttgtttctttacgtgctttataaatccgggaaacatttttacatcgttagaagtgcaagtcaagttatatgtatctataatgatttgtggaagttttgccacaacagttggtttatttacaatcttttttttattagccgtcaaatttctccctcatctcgtctctttatatctttattttatcataaaggagtatttgaaccgtcgcgcgggacaggtttatgatagggatattttattgaataattgatggtataagggactttcatgcttggaaagtggggattatgaattagatgcttgtccacaatatgttccaatgtaattaaaatttatgttcccaccttgaccaaacatcacgtccatacttaaatcgtccctcctttaataggtaaaatattaatttaccttaataaaaaaataatggataatagtattcgtctgaatttatataatcagggggaactattgatgctggggatactatttacagcggcgccatctactgatgtcgtaaaggatttgcaagataaagttatatcattgcaggatcatgaggtagcgtttttgaacaccacgatatctaatatgttgatcccc +pAMbeta1: + description: + prefix: GTCT + suffix: AAGC + tags: [origin] + sequence: gttaactaactcaacgctagtagtggatttaatcccaaatgagccaacagaaccagaaccagaaacagaatcagaacaagtaacattggatttagaaatggaagaagaaaaaagcaatgacttcgtgtgaataatgcacgaaatcgttgcttattttttttaaaagcggtatactagatataacgaaacaacgaactgaatagaaacgaaaaaagagccatgacacatttataaaatgtttgacgacattttataaatgcatagcccgataagattgccaaaccaacgcttatcagttagtcagatgaactcttccctcgtaagaagttatttaattaactttgtttAaagacggtatataaccgtactatcattatatagggaaatcagagagttttcaagtatctaagctactgaatttaagaattgttaagcaatcaatcggaaatcgtttgattgctttttttgtattcatttatagaaggtggagtttgtatgaatcatgatgaatgtaaaacttatataaaaaatagtttattggagataagaaaattagcaaatatctatacactagaaacgtttaagaaagagttagaaaagagaaatatctacttagaaacaaaatcagataagtatttttcttcggagggggaagattatatatataagttaatagaaaataacaaaataatttattcgattagtggaaaaaaattgacttataaaggaaaaaaatctttttcaaaacatgcaatattgaaacagttgaatgaaaaagcaaaccaagttaattaaacaacctattttataggatttataggaaaggagaacagctgaatgaatatcccttttgttgtagaaactgtgcttcatgacggcttgttaaagtacaaatttaaaaatagtaaaattcgctcaatcactaccaagccaggtaaaagcaaaggggctatttttgcgtatcgctcaaaatcaagcatgattggcggtcgtggtgttgttctgacttccgaggaagcgattcaagaaaatcaagatacatttacacattggacacccaacgtttatcgttatggaacgtatgcagacgaaaaccgttcatacacgaaaggacattctgaaaacaatttaagacaaatcaataccttctttattgattttgatattcacacggcaaaagaaactatttcagcaagcgatattttaacaaccgctattgatttaggttttatgcctactatgattatcaaatctgataaaggttatcaagcatattttgttttagaaacgccagtctatgtgacttcaaaatcagaatttaaatctgtcaaagcagccaaaataatttcgcaaaatatccgagaatattttggaaagtctttgccagttgatctaacgtgtaatcattttggtattgctcgcataccaagaacggacaatgtagaattttttgatcctaattaccgttattctttcaaagaatggcaagattggtctttcaaacaaacagataataagggctttactcgttcaagtctaacggttttaagcggtacagaaggcaaaaaacaagtagatgaaccctggtttaatctcttattgcacgaaacgaaattttcaggagaaaagggtttaatagggcgtaataacgtcatgtttaccctctctttagcctactttagttcaggctattcaatcgaaacgtgcgaatataatatgtttgagtttaataatcgattagatcaacccttagaagaaaaagaagtaatcaaaattgttagaagtgcctattcagaaaactatcaaggggctaatagggaatacattaccattctttgcaaagcttgggtatcaagtgatttaaccagtaaagatttatttgtccgtcaagggtggtttaaattcaagaaaaaaagaagcgaacgtcaacgtgttcatttgtcagaatggaaagaagatttaatggcttatattagcgaaaaatcagatgtatacaagccttatttagtgacgaccaaaaaagagattagagaagtgctaggcattcctgaacggacattagataaattgctgaaggtactgaaggcgaatcaggaaattttctttaagattaaaccaggaagaaatggtggcattcaacttgctagtgttaaatcattgttgctatcgatcattaaagtaaaaaaagaagaaaaagaaagctatataaaggcgctgacaaattcttttgacttagagcatacattcattcaagagactttaaacaagctagcagaacgccctaaaacggacacacaactcgatttgtttagctatgatacaggctgaaaataaaacccgcactatgccattacatttatatctatgatacgtgtttgttttttctttgctgtttagcgaatgattagcagaaatatacagagtaagattttaattaattattagggggagaaggagagagtagcccgaaaacttttagttggcttggactgaacgaagtgagggaaaggctactaaaacgtcgaggggcagtgagagcgaagcgaacacttgattttttaattttctatcttttataggtcattagagtatacttatttgtcctataaactatttagcagcataatagatttattgaataggtcatttaagttgagcatattagaggaggaaaatcttggagaaatatttgaagaacccgattacatggattggattagttcttgtggttacgtggtttttaactaaaagtagtgaatttttgatttttggtgtgtgtgtcttgttgttagtatttgctagtcaaagtgattaaatagaattc +pBM300: + description: + prefix: GTCT + suffix: AAGC + tags: [origin] + sequence: caagtcctacatcaagtcctaactcgtcaaatagtacatcttcatatagagactcaaaatggttagaaaactgtagggcaaattgttcgagatgtttttggatattatagtagttatcactaaattcctgttttaaaaagatgtcggtaaagcaagtataaccgccgttacctagatttgatacaatacttccctttctacctattcttggatatattgaggtaattacccaatttccatttccatctttttgcacatgtagtctaaaatcatatacatttccagattttattttgcatgttataaacttctgaactaagtaagtaccctcgcttatctgactctcgataaactctattagttggttttcactaagttttaacttttgttcgtttttactaatctcataataattatccgttttctctacaaagaagatacctgcccctttatgtccagataaaggttttacgattatttttttatagcgttcaaccatatctaacaagaccttataactagttagttttgaagtaggaataagatactgggtaaatttcttagcctgattcaccttattataaacggttaatttatcaccaacggagtgactagtgaatgggataatcttgcttaattcatgagttatttttgccattttatcgtttacaggggcgctagcattataaataacatcagggtaagaatattttctctcaacccatttgcctttttcgtaggccattcctaaaataactttttttttcaaatccaccttccctggagtgaaataaaaaaagtctattccttcacctttggcaactatagcaaaagcataagctttttttgcatttattgggtctttacgatgatgaagcatgccgattaaagtcataaaaaccatcctttcccttactcttaattgtttaggtatattcattaatttcatatattatttgtttttgatgaaccctggttgtcctttttttatatcatagattaagagaaggaaatatatgtattttatatgatgaaatcttaggataggatgatagatgtttatggatactattgtacttattgaaacaaataaatctggatcgagtagagaagcaattaaagcagcaaacaaacttaatttctttacggttttgttaactcgcagaaaaaaatttcttgaggaacgaaatgaattcccagatgtacatcaaatgattttcactgatacaaacgattataataatttagttacaataataaaaaagttaaaaaaggaagggaaaaa +pBS72: + description: + prefix: GTCT + suffix: AAGC + tags: [origin] + sequence: gaattcatattacttagaggatactattgagagagctattgataagatggttgagacattacctgagagccaaaaaactttttatgaatatgaattaaaaaaaagaaccaacaaaggctgagacagactccaaacgagtctgtttttttaaaaaaaatattaggagcattgaatatatattagagaattaagaaagacatgggaataaaaatattttaaatccagtaaaaatatgataagattatttcagaatatgaagaactctgtttgtttttgatgaaaaaacaaacaaaaaaaatccacctaacggaatctcaatttaactaacagcggccaaactgagaagttaaatttgagaaggggaaaaggcggatttatacttgtatttaactatctccattttaacattttattaaaccccatacaagtgaaaatcctcttttacactgttcctttaggtgatcgcggagggacattatgagtgaagtaaacctaaaaggaaatacagatgaattagtgtattatcgacagcaaaccactggaaataaaatcgccaggaagagaatcaaaaaagggaaagaagaagtttattatgttgctgaaacggaagagaagatatggacagaagaacaaataaaaaacttttctttagacaaatttggtacgcatataccttacatagaaggtcattatacaatcttaaataattacttctttgatttttggggctattttttaggtgctgaaggaattgcgctctatgctcacctaactcgttatgcatacggcagcaaagacttttgctttcctagtctacaaacaatcgctaaaaaaatggacaagactcctgttacagttagaggctacttgaaactgcttgaaaggtacggttttatttggaaggtaaacgtccgtaataaaaccaaggataacacagaggaatccccgatttttaagattagacgtaaggttcctttgctttcagaagaacttttaaatggaaaccctaatattgaaattccagatgacgaggaagcacatgtaaagaaggctttaaaaaaggaaaaagagggccttccaaaggttttgaaaaaagagcacgatgaatttgttaaaaaaatgatggatgagtcagaaacaattaatattccagaggccttacaatatgacacaatgtatgaagatatactcagtaaaggagaaattcgaaaagaaatcaaaaaacaaatacctaatcctacaacatcttttgagagtatatcaatgacaactgaagaggaaaaagtcgacagtactttaaaaagcgaaatgcaaaatcgtgtctctaagccttcttttgatacctggtttaaaaacactaagatcaaaattgaaaataaaaattgtttattacttgtaccgagtgaatttgcatttgaatggattaagaaaagatatttagaaacaattaaaacagtccttgaagaagctggatatgttttcgaaaaaatcgaactaagaaaagtgcaataaactgctgaagtatttcagcagttttttttatttagaaatagtgaaaaaaatataatcagggaggtatcaatatttaatgagtactgatttaaatttatttagactggaattaataattaacacgtagactaattaaaatttaatgagggataaagaggatacaaaaatattaatttcaatccctattaaattttaacaagggggggattaaaatttaattagaggtttatccacaagaaaagaccctaataaaatttttactagggttataacactgattaatttcttaatgggggagggattaaaatttaatgacaaagaaaacaatcttttaagaaaagcttttaaaagataataataaaaagagctttgcgattaagcaaaactctttactttttcattgacattatcaaattcatcgat +pCG1: + description: + prefix: GTCT + suffix: AAGC + tags: [origin] + sequence: attgaaggaggtttgtggcaagattttgatttagatcgttagataaattaggggaacctttattcttttctcctcctagaaaaggacttgcatatgcaagtcctcattcttaatacatgttcaaattacaaaaaatcatagatatctgtctgatgtgatgctttctctgcttctatgattacctctgcagtggttggcatccgtctatccctaattaccatctcatacccgataacttttcgtcctttttttacagttgttaaatctatatcggcgttgaatttcatttctaattcttcgcatgccaccatgagagcattcctcttaaataaacctgcaggccatgacttttcttcacccaagaaccaatcttgccattcctctaaacttcccttaatggttgtatattctcgatttcctaagcgatatgcttcccaaagtttcatcaagatcaaagaatactttgatttcacatttcctaactcatccaaatggaaagagtagtagtttcttttcaactcaaaaactaaaggtccagcatcttcagagaatttaaactctacaaggccatcctcaataaatcttattctgctgaaaagttgagtcatgattattgaacgtctaccatcaggcaatgtttcaggaatatatagagctgtattctcattaagtctcgaaaaagcttctgcaatccgtctataattctgaccgtttttagttaagccaaagtgcttcataatgtcaagactatgaaccgtaaagacatcaccaactctactatctttcgttacagatgaaaaacagtagtctaaaaccttgtgttcaaaagcttttaaattaccaaatgctttggccaagtcattcgcctggacaactaaataattttgacgagaagctagttcttcataaatcttacttgattttctatttgataccatcttgtttaccttttactatgttttctattatgctatattctatcaatttttaattttatagtcaataatggggtacaaaatagtcatcaatggggtacaaaatagtcatcaatggggtacaaaatagtcatcaatggggtacaaaatagtcaataatggggtacaaaatagtcaattatttatctgtaaacccttgatattactgagtttctaaggtcgacaaagtaatataaagtaatataaagtaggttagaaagtaaattagaaagtatcttcttaagaaaaaaatctatattttatttttaaaaaataaaaaaatctttcattcagttcactattttctatctaaccactttaaaatataatggctcaaaatctctattctaatatca +pCM66: + description: + prefix: GTCT + suffix: AAGC + tags: [origin] + sequence: cgacttccggcaagctatacgcgccctagaattgtcaattttaatcctctgtttatcggcagttcgtagagcgcgccgtgcgtcccgagcgatactgagcgaagcaagtgcgtcgagcagtgcccgcttgttcctgaaatgccagtaaagcgctggctgctgaacccccagccggaactgaccccacaaggccctagcgtttgcaatgcaccaggtcatcattgacccaggcgtgttccaccaggccgctgcctcgcaactcttcgcaggcttcgccgacctgctcgcgccacttcttcacgcgggtggaatccgatccgcacatgaggcggaaggtttccagcttgagcgggtacggctcccggtgcgagctgaaatagtcgaacatccgtcgggccgtcggcgacagcttgcggtacttctcccatatgaatttcgtgtagtggtcgccagcaaacagcacgacgatttcctcgtcgatcaggacctggcaacgggacgttttcttgccacggtccaggacgcggaagcggtgcagcagcgacaccgattccaggtgcccaacgcggtcggacgtgaagcccattgccgtcgcctgtaggcgcgacaggcattcctcggccttcgtgtaataccggccattgatcgaccagcccaggtcctggcaaagctcgtagaacgtgaaggtgatcggctcgccgataggggtgcgcttcgcgtactccaacacttgctgccacaccagttcgtcatcgtcggcccgcagctcgacgccggtgtaggtgatcttcacgtccttgttgacgtggaaaatgaccttgttttgcagcgcctcgcgcgggattttcttgttgcgcgtggtgaacagggcagagcgggccgtgtcgtttggcattgctcgcatcgtgtccggccacggcgcaatatcgaacaaggaaagctgcatttccttgatctgctgcttcgtgtgtttcagcaacgcggcctgcttggcctcgctgacctgttttgccaggtcctcgccggcggtttttcgcttcttggtcgtcatagttcctcgcgtgtcgatggtcatcgacttcgccaaacctgccgcctcctgttcgagtcgacgcgaacgctccacggcggccgatggcgcgggcagggcagggggagccagttgcacgctgtcgcgctcgatcttggccgtagcttgctggaccatcgagccgacggactggaaggtttcgcggggcgcacgcatgacggtgcggcttgctatggtttcggcatcctcggcggaaaaccccgcgtcgatcagttcttgcctgtatgccttccggtcaaacgtccgattcattcaccctccttgcgggattgccccgactcacgccggggcaatgtgcccttattcctgatttgacccgcctggtgccttggtgtccagataatccaccttatcggcaatgaagtcggtcccgtagaccgtctggccgtccttctcgtacttggtattccgaatcttgccctgcacgaataccagctccgcgaagtcgctcttcttgatggagcgcatggggacgtgcttggcaatcacgcgcaccccccggccgttttagcggctaaaaaagtcatggctctgccctcgggcggaccacgcccatcatgaccttgccaagctcgtcctgcttctcttcgatcttcgccagcagggcgaggatcgtggcatcaccgaaccgcgccgtgcgcgggtcgtcggtgagccagagtttcagcaggccgcccaggcggcccaggtcgccattgatgcgggccagctcgcggacgtgctcatagtccacgacgcccgtgattttgtagccctggccgacggccagcaggtaggcctacaggctcatgccggccgccgccgccttttcctcaatcgctcttcgttcgtctggaaggcagtacaccttgataggtgggctgcccttcctggttggcttggtttcatcagccatccgcttgccctcatctgttacgccggcggtagccggccagcctcgcagagcaggattcccgttgagcaccgccaggtgcgaataagggacagtgaagaaggaacacccgctcgcgggtgggcctacttcacctatcctgcccggctgacgccgttggatacaccaaggaaagtctacacgaaccctttggcaaaatcctgtatatcgtgcgaaaaaggatggatataccgaaaaaatcgctataatgaccccgaagcagggttatgcagcggaaaagatccgtcgaccctttccgacgctcaccgggctggttgccctcgccgctgggctggcggccgtctatggccctgcaaacgcgccagaaacgccgtcgaagccgtgtgcgagacaccgcggccgccggcgttgtggatacctcgcggaaaacttggccctcactgacagatgaggggcggacgttgacacttgaggggccgactcacccggcgcggcgttgacagatgaggggcaggctcgatttcggccggcgacgtggagctggccagcctcgcaaatcggcgaaaacgcctgattttacgcgagtttcccacagatgatgtggacaagcctggggataagtgccctgcggtattgacacttgaggggcgcgactactgacagatgaggggcgcgatccttgacacttgaggggcagagtgctgacagatgaggggcgcacctattgacatttgaggggctgtccacaggcagaaaatccagcatttgcaagggtttccgcccgtttttcggccaccgctaacctgtcttttaacctgcttttaaaccaatatttataaaccttgtttttaaccagggctgcgccctgtgcgcgtgaccgcgcacgccgaaggggggtgcccccccttctcgaaccctcccggcccgctaacgcgggcctcccatccccccaggggctgcgcccctcggccgcgaacggcctcaccccaaaaatggcagccaagctgacccgctagggacgtgaagtcgattccttcgatggttagcaatcaaagactcagagtgctgtctggagcgtgaatctaacggtacgtatctcgattgctcggtcgctattcgcactctgcgaaagttcgtaccgctcattcactaggttgcgaat +pGRB1: + description: + prefix: GTCT + suffix: AAGC + tags: [origin] + sequence: GATCTactcatcgacatcccaatctgctgtagctcacctacatcgctctcatcgcgtccgctctacgacccacttcttcaacccactccgtgggcaaggccagaccgccgtctgagcgtcttaaccacccggtggcggcgttcgcaacctcctgtacgggtcgctccgtggctccgacgggtccacctcgatgatgtccccgagatccccctccggcgcgatgcccaccatctcccactcctcgatgctgtcgccctcgtcgtcctcctccggttgcatcagctcctgtgcgccgttgctcggacggaaccactgccgaccactcgcccacatcgtcgcgtagaatgcacggacgttctcaggcatctcagacggctctgagccgtactcgccggccatgtacgccgccaggtacgccccgaggttctcgacgccaccgctgcgcgacgggtgcgacgagcgccgaactctcaccgcgtcctcgtcgccgttctcgtcgaacacctgatgggcgtcctcgcccgccgtcgggcagtttcgcagatgcgcgtccagcaccggctcgaactgctcggcgaccaccggccccctcacaaacacgccaagatggatgtgtacgtacccgctctcgtgcggttcaaggatggcgaggtactcccactctctaccctccaaaactcgcgccagcgcccgcctaacagcctcccacgactccagcaggtcctcgaagtgttccagcggaggccgcagccgtccctcgtcgtccgtactcgacgccgtgagcgtcaccatccccgtgtgcagaaggctcccccaccgctcccgaaggccccgttcgaggctctgcgccttcgcgtacagcatctcgcggtactccggcgtgaagcggttcggcatgtcaaagcccaccgtctcgcccagctcgttctcgacgacgatctgcgttccacgctggtcgtcgtaccagctctggaagcggtcgatggcctccccccacgacaccgccttgacggcccggctcgtgtcgaaggtcagttcatcccgcagtttcatcccgtctcgtttcgccatcggcacgtgtgaggcgctgaggctgtcgttctccgggcctgcacggttattcaaaggttgccgagaaccgcccgagttggcaggccggtcgggcggttcacccggctgattaggtgtgctcaccggaacccctccgttccgggaacgaggacggccgcgttgcggcctgtgttccgggctgcgggccgctgcgcggcccgttccgggctttcacccatcccttcactgcggggatgctggacagcacgggagatcccccgcctgccggctgttgcggcccctgagagggacctggggcacatataccagccgtctcgccgtcgggaaagacctaagcggacggacgaagaagggccgagtgaaccagccaggcaaggtggttcgtttgccggttgcaggccgcgggtacggccccttccggttttgtcccggcccttctccggggcacctttcacgttgcgtagctgttactccaccgtcattcagctatccagttattagttgcggcgtggagccagtattggtccttaat +pIP404: + description: + prefix: GTCT + suffix: AAGC + tags: [origin] + sequence: gaattcttttaaggggcttatagattatctattttaaataaaaaggattcttttaatttattattaaaagaatcctttttattaattatttttaactgaattaatgtatatatatatatgataaaatccaaatgtaaaaatgcaagaaaaaacatacattaaaaagcttataatataatattgtggaatattgaaattaattttactaaaaactattccaatcaaggcagcaataataaaaatagttttatttattggtttagagtacgtgtttttatttttaagtttctttttaatagctgttagaaagaaacttaaaaataaaatatatataataataaatataataaaaatatatataaatttagaaaaagataatatttgataaaacatatttaaagatataatcaaatcctgtaaagatgaaaatgaaaataaaattcctaaaaataagtttaataaagccggaaatttcactagtttttttaaaaacaataaatataataaagatattattataaaacttaaatttaataataacatcataaggaaaatggattggtttacttcaacggctttattatatgaaaaaaatacaattaaatttaaacataatattacaaaaagtattgttaaaattgaaaaaattaaatctttagtatctttggaaataatcataatttataagctcctttgattttttatataaattatacattaattatttaatatataaaaactaaaattagtaaatcatctaaatattacaaatgaagaaatattatttgtaaataattatagtaatatattatggaatgaaatagtaaaaaattaaaaagaatcatttaatttagtttaaaatattttaataaaaataataaattaaaacataaaaaatagattaagtttcaattggggacttactctattttttattaagagtttaagaatagttaataattattgaaaatatgatataattttcttagttttacagaaggggaggtgaataagtgagaaacgcattaaagctgaataaaaagaactatatagataatattcatagtagaagtaaagggtggataaccaggagcgttatagataaaaaaggatatagccaatggcactataaatacgctgaattaaaagatttagatatgagtgatgaaaatatctatataactctaaataccttttataagccgtgtaggcgattagaaaatataaaagagttaaatacactgtttatagacttggattattataaaactggcaaaactaaagaccaggtattaatggacttagaaaagaattattttaatcaaagtattcctataccaaactatgtaatagatagtggaagaggaatgtatttaatatggataataaatgcagtacctagtaaagcattaccattatggaaagcggttcaagaatatttatataatcaattaaaatactttggagcagatagacaagccttagatgcaaccagaatattaagagttccaggaagtataaactctaaatcaaaaacagtagtcaatatattagatgagtacgaatatatttatgacttaagagaaatacaaaatggatttttacctgaattaaaaccatatgaaaggaaaaagggtagaccaagcaaaataaattatatttatagagaaagaagtttatattatggaagaatacaagacataataaaactttgtgaactaagagaatatgatttaaaaggacacagagagcttatattatttttatatagatattatctttgtagctttacagaggacattgagaaggcattaaatgatgttttagaacttaatagtatgtttagacaacatttaagtgaaagagaagttataagagcaactagaagtgctgaaagatgttatttagataaaaataagcaatataagtataagaatgaaactcttatagagttattagaaattacagaagaagaacaaaaatatatgacaataataatttctaaaaaggaatataagagaagagaaaatattagaggtaaaaaaaattatcaagagcaattaaaagctaaaggaaaagcaacaaaaaaagaggaattaaatgtattaagaaaaaaaataaaagcccttaaagaaaaaggctttaaaaataaagaaattactctaatgttagaagtaccaataaaaacattagaacgtcatattacgtatatgaaaaaaaatgggcttttataaaggctcattttttatattcttttcttcaaagattatataatataaaaaaatttttttcaaactttaaataaaaaatatttttatatttttttatttttttatttttatatttttttatttttttatttttatatttttttatttttatatttttttatttttatatttttttatttttatatttttttatttttttatttttttatttttttatttttttatttttttatttttttatttttttattttttttatttttttatttttttatttttttatttttttatttttaccctcatttttttacgcttgtattatagggtactttgtacctgttcttttttttggggaggttgtaaagataattttttactttagttagttccgaaggaacttttattatagtgcccctttatttttttgcgtggcattgaatgtaaaaaattatcactatactagggcgtaaagtaatattacatgtgtctcaaagtgggattaaagcgggattttatagggcgtgtttgtggcttagagtgggattattggaaatttttttgatatc +pLS20: + description: + prefix: GTCT + suffix: AAGC + tags: [origin] + sequence: ATCATTACTAcaggcccggggctttacgtcaaaaaaaaaatcgtgtaagataaagtcaagttaagtcaatgtgaaagttttggcattgaattctggttctagttattctaaataaacaacaaaaaaccctgttgtttgcagacaacaaggttaatagatttgatgatacatattttgtttgcgagctatttgcaaaattgaatacaaataaagaatgacaaataaaaagtgtccgatgttgcagcatcgaacacttggtttgtaagccctcacttacaatgtttaagttatAtcttcattttaccgaaaacaagtgtatacgtcaacacttttcggctatttttggacgacagaaaaccactgtttggagggtatcaaaccgtggttttctgtcgttttttgtttgtcacccagctgccgttaaggggtgtaaaataaactgacgtcaggagttgccccgtccaccgcaaacccggggatataccacaataagcgttccttgctgcgcttgtggtgaatggcgaggacggccattaacggtccggagaatgtggataaaccactaggttgcttggcgcactacggtgcgggcaactatggccaagcttctatgcagcggcacacaattgagtgtgaccgtgtgagagaacacgagacaaaccccatagaagttgcatacaggacaagcattgtatgtaaaaaggtcatggcaggcgaaagcctggcgatagcaaggcagagctggaaacctcagaTgtctctatgacgctagtttcaaatcctgatcgattgcctatgtctgtccgtttttattttttgcgcgacagacataggcaatcgaaattcgccctgtttcctagaacagtcagctattctgctagtttcaagttctcaaagtcaagagaaaaacactaaataaaatgcgaatttccttaaaacccagaggttttaagttcctttttaagggttgttcgtttgatagaacattagtattttaatatatttttcctgtgggatttcactaacgtatactgttattccctttggcctatcttaaaaacttgtatagattgtgtttgagccgtttagtatgatatcagtacatgcttggg +pMflST: + description: + prefix: GTCT + suffix: AAGC + tags: [origin] + sequence: CCGTGAACACGAGCGTGTTTAGTTTTTGATGGTTGTCAAGTTCTTTTCATtgtattcaaacccccttttaaagttgataattcttgttaattataatataagtgctggaaaaagtatagttttttattgttatttttaagaaaatttatccattgtgtttttatatgaactttttcaacattttttataaccttttgtggaaaagcggaaaaaagatatataaatactaaactattgctttttagtgttattcattatgtttatttctttttaaaatatttttttatccacacattgtggaaaagttttattattatgtgttttaatttattttatgaaggggaattaatatggaaacaaaagcgctttgagaaaaattaattaataagttaaagaaggaaaaattaatagaccaagacattattgaagaacatattgttacttcagaattaattaaaatttcaaatacagagttcgtaatccttgttagaagtaatcttggtgtaactattttaaatgaatttaaagaagtatttgtgtacgaatttaaataagttttaaacagttatgtgtctgttgattttttaactaaagaaatttttgagaaaaatacaaaaaaagagaataaaaaagaacccataaatactgttttatctgaaaatgcacttacttttgagaattttatagttgggtcaagtaataaacaagctaatttagctgctaaaaatgttgttgcaaaccctggtatgagttttaatcctttatttatttatggagattcaggactgggtaaaacacatttattacaagctattaaaaatcaagcagaattaaatggaaaaaaagttttatatttaacatctgaagaatttacaaaaagaattgttaatgctttaaacaaaggtgatttaagtgaaatagaagaattaaaaactgagataaattctaatgaattctttattttagatgatgttcaatttttaagtaaaaaagataaaacaaacgagtttttctttaacattattaacaattttactgaaaatggtaaacaattagttttttctagtgataagactcctgaattgttaaatggttttgataaaagaatgataactagatttaattcaggtttatcaacacctattaatgctttagatattccaactgccaaattgataattgaagctgaaattaaaaaacaaggtttaaaacaaaaaattaaagaagatgccgttgtttacttagctcaaaactttagcgacgatgttagaaaaattaaaggtttagttaatagattacttttctttggtattcaaaacgatttaggtcacataattgatttggaagatgttattgacttatttaaagatacaccttcagctaatttaggattattaaatgttaaaaaaattaaagaagttgttgctaaaaaatatgatgttactataaaagccatcgatggtaaagcaagaacaactgctataaaaaatgctagacatctttctatgtattttgcaaaaataattttaaaccatacatcaactcaaattggtgcagaatttggtggaagagatcatagtacagttttaagtgctatttcgcgtattgaaaaattaatatataaagaaaaagaattcaaaaaaatagttgagtctttaaaaaacgaaataataggaaaatagatgtaaattcgcatctattttttttgtgtggaaaagaaattaaacttgttgaataaaaagtcatttaaacctttatttaccgttgttttatgagtgttttccacatatcaactgcattataaatattacttatataaagaataatataatatataaaagcatgcgtatttcttttgtttttattttataaatatgagataatatatattgtaaataagaggtgttatatgag +pMK3: + description: + prefix: GTCT + suffix: AAGC + tags: [origin] + sequence: aataaaaaaagcacctgaaaaggtgtctttttttgatggttttgaacttgttctttcttatcttgatacatatagaaataacgtcatttttattttagttgctgaaaggtgcgttgaagtgttggtatgtatgtgttttaaagtattgaaaacccttaaaattggttgcacagaaaaaccccatctgttaaagttataagtgactaaacaaataactaaatagatgggggtttcttttaatattatgtgtcctaatagtagcatttattcagatgaaaaatcaagggttttagtggacaagacaaaaagtggaaaagtgcgtccatggagagaaaagaaaatcgctaatgttgattactttgaacttctgcatattcttgaatttaaaaaggctgaaagagtaaaagattgtgctgaaatattagagtataaacaaaatcgtgaaacaggcgaaagaaagttgtatcgagtgtggttttgtaaatccaggctttgtccaatgtgcaactggaggagagcaatgaaacatggcattcagtcacaaaaggttgttgctgaagttattaaacaaaagccaacagttcgttggttgtttctcacattaacagttaaaaatgtttatgatggcgaagaattaaataagagtttgtcagatatggctcaaggatttcgccgaatgatgcaatataaaaaaattaataaaaatcttgttggttttatgcgtgcaacggaagtgacaataaataataaagataattcttataatcagcacatgcatgtattggtatgtgtggaaccaacttattttaagaatacagaaaactacgtgaatcaaaaacaatggattcaattttggaaaaaggcaatgaaattagactatgatccaaatgtaaaagttcaaatgattcgaccgaaaaataaatataaatcggatatacaatcggcaattgacgaaactgcaaaatatcctgtaaaggatacggattttatgaccgatgatgaagaaaagaatttgaaacgtttgtctgatttggaggaaggtttacaccgtaaaaggttaatctcctatggtggtttgttaaaagaaatacataaaaaattaaaccttgatgacacagaagaaggcgatttgattcatacagatgatgacgaaaaagccgatgaagatggattttctattattgcaatgtggaattgggaacggaaaaattattttattaaagagtagttcaacaaacgggccagtttg +pMTLB: + description: + prefix: GTCT + suffix: AAGC + tags: [origin] + sequence: Aaattcatattacttagaggatactattgagagagctattgataagatggttgagacattacctgagagccaaaaaactttttatgaatatgaattaaaaaaaagaaccaacaaaggctgagacagactccaaacgagtctgtttttttaaaaaaaatattaggagcattgaatatatattagagaattaagaaagacatgggaataaaaatattttaaatccagtaaaaatatgataagattatttcagaatatgaagaactctgtttgtttttgatgaaaaaacaaacaaaaaaaatccacctaacggaatctcaatttaactaacagcggccaaactgagaagttaaatttgagaaggggaaaaggcggatttatacttgtatttaactatctccattttaacattttattaaaccccatacaagtgaaaatcctcttttacactgttcctttaggtgatcgcggagggacattatgagtgaagtaaacctaaaaggaaatacagatgaattagtgtattatcgacagcaaaccactggaaataaaatcgccaggaagagaatcaaaaaagggaaagaagaagtttattatgttgctgaaacggaagagaagatatggacagaagaacaaataaaaaacttttctttagacaaatttggtacgcatataccttacatagaaggtcattatacaatcttaaataattacttctttgatttttggggctattttttaggtgctgaaggaattgcgctctatgctcacctaactcgttatgcatacggcagcaaagacttttgctttcctagtctacaaacaatcgctaaaaaaatggacaagactcctgttacagttagaggctacttgaaactgcttgaaaggtacggttttatttggaaggtaaacgtccgtaataaaaccaaggataacacagaggaatccccgatttttaagattagacgtaaggttcctttgctttcagaagaacttttaaatggaaaccctaatattgaaattccagatgacgaggaagcacatgtaaagaaggctttaaaaaaggaaaaagagggtctgccaaaggttttgaaaaaagagcacgatgaatttgttaaaaaaatgatggatgagtcagaaacaattaatattccagaggccttacaatatgacacaatgtatgaagatatactcagtaaaggagaaattcgaaaagaaatcaaaaaacaaatacctaatcctacaacatcttttgagagtatatcaatgacaactgaagaggaaaaagtcgacagtactttaaaaagcgaaatgcaaaatcgtgtctctaagccttcttttgatacctggtttaaaaacactaagatcaaaattgaaaataaaaattgtttattacttgtaccgagtgaatttgcatttgaatggattaagaaaagatatttagaaacaattaaaacagtccttgaagaagctggatatgttttcgaaaaaatcgaactaagaaaagtgcaataaactgctgaagtatttcagcagttttttttatttagaaatagtgaaaaaaatataatcagggaggtatcaatatttaatgagtactgatttaaatttatttagactggaattaataattaacacgtagactaattaaaatttaatgagggataaagaggatacaaaaatattaatttcaatccctattaaattttaacaagggggggattaaaatttaattagaggtttatccacaagaaaagaccctaataaaatttttactagggttataacactgattaatttcttaatgggggagggattaaaatttaatgacaaagaaaacaatcttttaagaaaagcttttaaaagataataataaaaagagctttgcgattaagcaaaactctttactttttcattgacattatcaaattcatcg +pNG168: + description: + prefix: GTCT + suffix: AAGC + tags: [origin] + sequence: accatagagttgagtagaatatttgacgaatgggaaacacatcatcgaaatttggtggttctacactgtgcgggatcgtcagacaccgactgttacctttaagttgcatccgtgaaaacaaggggccaaggcaggacagagtaatcactggtcggcgttggaagcctgaagaaggccccggacacgaacctctcgtgtccccacaggttgctacctcggccttcttccgctgtgggcttaagcgttaccccacgtagccgctgcgctgccaaatgaaatgtcctcatcacgatgacagcgatgcaaactacacagacacaccaagaccgaggtgaccgcgagtagctgacgctactccagaccaatgactcacaggtacagactcaaacgacaatccgggtccgatgctgccaggcttcgccccggatgtcgttgtgaagcatcgtcgcagcggaccagcagagccggtcctcaacacggatggagcggtattatcagtacgctcctatctacaatgtggacgcgaggggtcaaattggtttcgggaccctcgcatttgccggcttccacgcagtccgcacttctccacggactgcatgcacacgcctaatcaacaacaaggaatccggaagatcgttccaggagggaccctctctactgctggaattaccattaccgaggtcaccccacgggtaacagaatggatacctgaccttcttgaggaactactgccacggagtattcaatcggttcgaaaattcatccggcaagaggacccagaagtcctcacgcacgcacgatacaacaccgtctatcgccgacttcaagaggaaaccctgaggttcgaccatcaagaatggtgctccacaacggatatttggagtgatgcagaggctgaagcggttgaatacgtagaatcacttgtcgagttcgcagtcaaatattctgacgttgacgaagatgatctcgacgaactttcggagtatcaccagcagcggtgcaaatcgcttaaacagactctcactacgatcagtaccggacgtggcccactcaacgctggccttgaagccctcgccaagggacccgtacgactccacgatgagctcgatgacgcaccgcaaccgatcacgcttgtccttgatggcgagttgtggtcaaaactcgacgatagaggaacaggtatccgagcacttgcagctatcgccgtgctcggctccaccttcgacgtccgcctggttatttcaccagcgttagacgccgcgattgagcgacggtatccagactggtatgactcccatctccgtcttactgaaacccgtgaaacctcctctgtagaatcagcgggtggcgacggacaaccatcggcggagcagctcgaggaagcatgggaagcaatacagaacctcccagaggaatcagggagactccgacttctccggaacctccctatagagggctctcgagactatcgtgatctcaagcaggatgatgagattgacgtgcaggcagggactgtcggccgttacatcctcgatctagaagaattagggcttgtggatatcgaccgtcgtggacaatacaacagcgcatccctcaccggcttaggacaagtagcagttgagcagtatgtcaccacggactaccgggtgatccatccgacccaatcgacgctggaaacgcatcttacgccgacccctcagccccaagcaagtacagtgtatcccgcgcgatcggacacgagggagggggatcagcctgggacagcggaggattggatagctgcgacaggcagtcctagtgagggtgctgactacgttcaatggctcgatgggccgtctggtgttctcgacgcttggggaatgcatcagcggtaccttgctggccgtcgagatcgtggtgtcaccctagtcgatgaccgtatcgagcgtttcgaggacggtcgcgtatcgtatctcagttgcttcgacgatgatctcttcgtagccactcagtggggcggcccccttcctaccctgggacgtattgcgggtgccttactcagcgacaaagcactgagtaagatccttaccccttctcgactcggaaaccagttcgaagaaatcaacgatgcggtcgttgaacaactcgatagagaagctggggaaatcatccgtagaggacaccaaatcggctggttcagcgaggacgaagaggactacgatggttggcgtgagcgtattggctccgttcgcagtctgtgtttgcaacaagttggtgaactcacaaacagtgacgacgtcgaagcccggactgagttactacgtgacctgcacggcttggttgcctcagcgacacagctgtactacgcagccggtgtagacgtcacaatcaacgttcgagtccctgacacgggaatgctcatctcggatgaacgccgactcgacgatttcctcggtttcgcccggtacaccattccgaaacagtccgtgtacgggattcattcggggtataggatgctcctcgaggatcgaccggagaaactcaaacgacgacttccctacgaggtggatgatgctgattccacaatgcatctgaccgcatcgtgggttttctccggatcaacgatgatcgatcttcacgatgacatcgaggacgccattgaaatggaaaccaacgaaattcgtgaagcaattgctaatggacaggagtcagcaccggtaatggaaatccccgtccagattgggaactcctactcagcaattcgtaatcacgtcgaggactacgcttcagcgaagaactaccaggtagctcaccaggaggatattcacgaaggaaaacaggatctcgagcgactcgtccgcctgttcctccgtgtcctcgggacagaggatcgaccacatcgagcgtgtcctcacgacgttgccgaggcaatgcttcatgttgctcaatcctcccggaactacgatttcatcaccgtgcgagacatctcgtacggactgtcgaatctccccacgaaacgactcttacccgagctcccaccaacagcaacgaagctcctcaaaaccctgcttgatgcggatgacccgatgggacggtctgaaatcatcgataccgctgacatttcggaaagtagctatgatcgctacatcaacgaactcgccgcttgggatatcattgaaccccgcgaaatcgagggacaccgtcggtgggaagctcacttagagccgtggtggacaccgcagagtgatcgtgacgaaccctatgccgaccctgaccccgacacgggaatactgtacgctgaatttccccgtgatgtcgctagtgcggtgatgtgccacctcatcacccactacgacttacccgaccttgagacagcgtatcttgagggtatccaaccgggggacgacatcaaggccctcttcgatgatcacgaccgactcagacgatggcggccattcctctggggtgcattcgccgactcggacaaacttgaaagaggcccatctggtacagcagcttcggactcgaccgtagttcgactcggtcaatctccaggacccgacacagcacaatcgagcttccaagatgtctcagaaacagcaacccaacgagatcgactcagtcaaccgtcgccagggctggactaatgattgacggcaatcgataactacgcaatgaatcctccacaaccagcatctgaaccgtactcgaaaggtgatgcagtaaccgtatacgtgggc +pSK41: + description: + prefix: GTCT + suffix: AAGC + tags: [origin] + sequence: gatttatcggcctcataattttagatatattaattatattagagatttgttatcaaatacatttcattacaagcatataattgacctgtgaggcaacaaaaaatttttaatttttttaaaatagtgttactattgaacatagaaaaagacacctaacgcttggcggttgttagatgtctaaagagtttaaataatatgtgttatttacttttacaatttgtttgtatctttattatatatacaacctcgatacattacaagataaaagcaacaagtctttaagttatatataacggataacattatataaccttattcatcacttgtaaataacgatattacttaaactccttaaatatacatttaaaggagtttttatcatgtctaaacaattttttacagtagaagaaaattataaagaacgcttttaccaattacctaaagtattttttactaatcctaattataaagatttatcgaatgacgctaaaattgcttacgcaatattgagagatcgtttacaattatctattaaaaataactggatagatacagaaggtaacatatatttcatttatacagttgctgatttagaagtgatattaaattgtgggaacaaaaaaatcactaaaattaaaaaagaattagaaaatgtagatttactaatccaaaaaaggcagggtttaaacaaaccaaatctactttatttattaaaaccagccataactaaaaatgatatttatgaaatagataaggctgaaaatgaagttgaggccttacaagacaaggaagtgtcaaaaggacacgtccagaaatgtcaaaaggacacgtctagaaatgtcaaaaggacacgtctagaaatgtcaaaaggacacacaaatgatactgattttattgatactgattttattgatactgaaagtaatgatatgaataatatgaatgatacaaatcaacattcaaatcattcaaatcacttttcaaatattcatgataaagaatctcttaaatatatagaacttcaagaactccccgaattaattaaaagttatattaataatttcagctatgaagaagttaaaagtattaaatctgttattttaaaagctaaaaaatcatttaataacaaatacgatactttctatatgttagaagatatagatgaagaattactattggttttaaaacgatttaaaggttatcttgttaaaaaacaagaaaaagtagctaatatggaaggttatttaatgagaagtatcattgctgaacttgaagaaatgcactcaactattatgagaagaaaaaatatggaaaacaatccattatctctatttaattagtttttgaagttaaattttcaaaaaaaagagattatacacctaaaaaggtgtataatctttac diff --git a/parts/parts/ecoli/ecoli.yaml b/parts/parts/ecoli/ecoli.yaml new file mode 100644 index 0000000..a31b528 --- /dev/null +++ b/parts/parts/ecoli/ecoli.yaml @@ -0,0 +1,103 @@ +# CIDAR +eBFP2: + description: EBFP2 is a basic (constitutively fluorescent) blue fluorescent protein published in 2007, derived from Aequorea victoria. It is reported to be a rapidly-maturing monomer with moderate acid sensitivity. + prefix: + suffix: + references: ["https://www.fpbase.org/protein/ebfp2/"] + sequence: atggtgagcaagggcgaggagctgttcaccggggtggtgcccatcctggtcgagctggacggcgacgtaaacggccacaagttcagcgtgaggggcgagggcgagggcgatgccaccaacggcaagctgaccctgaagttcatctgcaccaccggcaagctgcccgtgccctggcccaccctcgtgaccaccctgagccacggcgtgcagtgcttcgcccgctaccccgaccacatgaagcagcacgacttcttcaagtccgccatgcccgaaggctacgtccaggagcgcaccatcttcttcaaggacgacggcacctacaagacccgcgccgaggtgaagttcgagggcgacaccctggtgaaccgcatcgagctgaagggcgtcgacttcaaggaggacggcaacatcctggggcacaagctggagtacaacttcaacagccacaacatctatatcatggccgtcaagcagaagaacggcatcaaggtgaacttcaagatccgccacaacgtggaggacggcagcgtgcagctcgccgaccactaccagcagaacacccccatcggcgacggccccgtgctgctgcccgacagccactacctgagcacccagtccgtgctgagcaaagaccccaacgagaagcgcgatcacatggtcctgctggagttccgcaccgccgccgggatcactctcggcatggacgagctgtacaagtaa +E1010m: + description: Fluorescent protein. Red. Modified from Bba_E1010 to fix illegal sites. + sequence: atggcttcctccgaggatgttatcaaagagttcatgcgtttcaaagttcgtatggaaggttccgttaacggtcacgagttcgaaatcgaaggtgaaggtgaaggtcgtccgtacgaaggtacccagaccgctaaactgaaagttaccaaaggtggtccgctgccgttcgcttgggacatcctgtccccgcagttccagtacggttccaaagcttacgttaaacacccggctgacatcccggactacctgaaactgtccttcccggaaggtttcaaatgggaacgtgttatgaacttcgaagatggtggtgttgttaccgttacccaggactcctccctgcaagacggtgagttcatctacaaagttaaactgcgtggtaccaacttcccgtccgacggtccggttatgcagaaaaaaaccatgggttgggaagcttccaccgaacgtatgtacccggaggatggtgctctgaaaggtgaaatcaaaatgcgtctgaaactgaaagacggtggtcactacgacgctgaagttaaaaccacctacatggctaaaaaaccggttcagctgccgggtgcttacaaaaccgacatcaaactggacatcacctcccacaacgaggactacaccatcgttgaacagtacgaacgtgctgaaggtcgtcactccaccggtgcttaataa +E0040m: + description: Fluorescent protein. Green. Modified from Bba_E0040 to fix illegal site. + sequence: atgcgtaaaggagaagaacttttcactggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggagagggtgaaggtgatgcaacatacggaaaacttacccttaaatttatttgcactactggaaaactacctgttccatggccaacacttgtcactactttcggttatggtgttcaatgctttgcgagatacccagatcatatgaaacagcatgactttttcaagagtgccatgcccgaaggttatgtacaggaaagaactatatttttcaaagatgacgggaactacaagacacgtgctgaagtcaagtttgaaggtgatacccttgttaatagaatcgagttaaaaggtattgattttaaagaagatggaaacattcttggacacaaattggaatacaactataactcacacaatgtatacatcatggcagacaaacaaaagaatggaatcaaagttaacttcaaaattagacacaacattgaagatggaagcgttcaactagcagaccattatcaacaaaatactccaattggcgatggccctgtccttttaccagacaaccattacctgtccacacaatctgccctttcgaaagatcccaacgaaaagagagatcacatggtccttcttgagtttgtaacagctgctgggattacacatggcatggatgaactatacaaataataa +E0030: + description: Fluorescent protein. Yellow. + sequence: atggtgagcaagggcgaggagctgttcaccggggtggtgcccatcctggtcgagctggacggcgacgtaaacggccacaagttcagcgtgtccggcgagggcgagggcgatgccacctacggcaagctgaccctgaagttcatctgcaccaccggcaagctgcccgtgccctggcccaccctcgtgaccaccttcggctacggcctgcaatgcttcgcccgctaccccgaccacatgaagctgcacgacttcttcaagtccgccatgcccgaaggctacgtccaggagcgcaccatcttcttcaaggacgacggcaactacaagacccgcgccgaggtgaagttcgagggcgacaccctggtgaaccgcatcgagctgaagggcatcgacttcaaggaggacggcaacatcctggggcacaagctggagtacaactacaacagccacaacgtctatatcatggccgacaagcagaagaacggcatcaaggtgaacttcaagatccgccacaacatcgaggacggcagcgtgcagctcgccgaccactaccagcagaacacccccatcggcgacggccccgtgctgctgcccgacaaccactacctgagctaccagtccgccctgagcaaagaccccaacgagaagcgcgatcacatggtcctgctggagttcgtgaccgccgccgggatcactctcggcatggacgagctgtacaagtaataa +cre: + description: cre/lox + sequence: atgtccaatttactgaccgtacaccaaaatttgcctgcattaccggtcgatgcaacgagtgatgaggttcgcaagaacctgatggacatgttcagggatcgccaggcgttttctgagcatacctggaaaatgcttctgtccgtttgccggtcgtgggcggcatggtgcaagttgaataaccggaaatggtttcccgcagaacctgaagatgttcgcgattatcttctatatcttcaggcgcgcggtctggcagtaaaaactatccagcaacatttgggccagctaaacatgcttcatcgtcggtccgggctgccacgaccaagtgacagcaatgctgtttcactggttatgcggcgaatccgaaaagaaaacgttgatgccggtgaacgtgcaaaacaggctctagcgttcgaacgcactgatttcgaccaggttcgttcactcatggaaaatagcgatcgctgccaggatatacgtaatctggcatttctggggattgcttataacaccctgttacgtatagccgaaattgccaggatcagggttaaagatatctcacgtactgacggtgggagaatgttaatccatattggcagaacgaaaacgctggttagcaccgcaggtgtagagaaggcacttagcctgggggtaactaaactggtcgagcgatggatttccgtctctggtgtagctgatgatccgaataactacctgttttgccgggtcagaaaaaatggtgttgccgcgccatctgccaccagccagctatcaactcgcgccctggaagggatttttgaagcaactcatcgattgatttacggcgctaaggatgactctggtcagagatacctggcctggtctggacacagtgcccgtgtcggagccgcgcgagatatggcccgcgctggagtttcaataccggagatcatgcaagctggtggctggaccaatgtaaatattgtcatgaactatatccgtaacctggatagtgaaacaggggcaatggtgcgcctgctggaagatggcgattag +araC: + description: Controller protein, araC repressor/activator (activates pBAD, I13453). Otherwise known as C0080 + sequence: atggctgaagcgcaaaatgatcccctgctgccgggatactcgtttaacgcccatctggtggcgggtttaacgccgattgaggccaatggttatctcgatttttttatcgaccgaccgctgggaatgaaaggttatattctcaatctcaccattcgcggtcagggggtggtgaaaaatcagggacgagaatttgtctgccgaccgggtgatattttgctgttcccgccaggagagattcatcactacggtcgtcatccggaggctcgcgaatggtatcaccagtgggtttactttcgtccgcgcgcctactggcatgaatggcttaactggccgtcaatatttgccaatacgggtttctttcgcccggatgaagcgcaccagccgcatttcagcgacctgtttgggcaaatcattaacgccgggcaaggggaagggcgctattcggagctgctggcgataaatctgcttgagcaattgttactgcggcgcatggaagcgattaacgagtcgctccatccaccgatggataatcgggtacgcgaggcttgtcagtacatcagcgatcacctggcagacagcaattttgatatcgccagcgtcgcacagcatgtttgcctgtcgccgtcgcgtctgtcacatcttttccgccagcagttagggattagcgtcttaagctggcgcgaggaccaacgcatcagccaggcgaagctgcttttgagcactacccggatgcctatcgccaccgtcggtcgcaatgttggttttgacgatcaactctatttctcgcgagtatttaaaaaatgcaccggggccagcccgagcgagttccgtgccggttgtgaagaaaaagtgaatgatgtagccgtcaagttgtcagctgcaaacgacgaaaactacgctttagtagcttaataa +luxR: + description: Controller protein, luxR repressor/activator (in concert with HSL, represses pLuxR(pR) R0063. Also up-regulates pLuxR(pL) R0062). Annotated as C0062. + sequence: atgaaaaacataaatgccgacgacacatacagaataattaataaaattaaagcttgtagaagcaataatgatattaatcaatgcttatctgatatgactaaaatggtacattgtgaatattatttactcgcgatcatttatcctcattctatggttaaatctgatatttcaatcctagataattaccctaaaaaatggaggcaatattatgatgacgctaatttaataaaatatgatcctatagtagattattctaactccaatcattcaccaattaattggaatatatttgaaaacaatgctgtaaataaaaaatctccaaatgtaattaaagaagcgaaaacatcaggtcttatcactgggtttagtttccctattcatacggctaacaatggcttcggaatgcttagttttgcacattcagaaaaagacaactatatagatagtttatttttacatgcgtgtatgaacataccattaattgttccttctctagttgataattatcgaaaaataaatatagcaaataataaatcaaacaacgatttaaccaaaagagaaaaagaatgtttagcgtgggcatgcgaaggaaaaagctcttgggatatttcaaaaatattaggttgcagtgagcgtactgtcactttccatttaaccaatgcgcaaatgaaactcaatacaacaaaccgctgccaaagtatttctaaagcaattttaacaggagcaattgattgcccatactttaaaaattaataa +tetR: + description: Controller protein, tetR repressor (represses pTet, C0040. can be inhibited by tetracyclin or aTc) + sequence: ATGtccagattagataaaagtaaagtgattaacagcgcattagagctgcttaatgaggtcggaatcgaaggtttaacaacccgtaaactcgcccagaagctaggtgtagagcagcctacattgtattggcatgtaaaaaataagcgggctttgctcgacgccttagccattgagatgttagataggcaccatactcacttttgccctttagaaggggaaagctggcaagattttttacgtaataacgctaaaagttttagatgtgctttactaagtcatcgcgatggagcaaaagtacatttaggtacacggcctacagaaaaacagtatgaaactctcgaaaatcaattagcctttttatgccaacaaggtttttcactagagaatgcattatatgcactcagcgctgtggggcattttactttaggttgcgtattggaagatcaagagcatcaagtcgctaaagaagaaagggaaacacctactactgatagtatgccgccattattacgacaagctatcgaattatttgatcaccaaggtgcagagccagccttcttattcggccttgaattgatcatatgcggattagaaaaacaacttaaatgtgaaagtgggtccgctgcaaacgacgaaaactacgctttagtagcttaataa +lacI: + description: Controller protein, lacI repressor (in concert with CAP, represses pLacI, R0010) + sequence: ATGatggtgaatgtgaaaccagtaacgttatacgatgtcgcagagtatgccggtgtctcttatcagaccgtttcccgcgtggtgaaccaggccagccacgtttctgcgaaaacgcgggaaaaagtggaagcggcgatggcggagctgaattacattcccaaccgcgtggcacaacaactggcgggcaaacagtcgttgctgattggcgttgccacctccagtctggccctgcacgcgccgtcgcaaattgtcgcggcgattaaatctcgcgccgatcaactgggtgccagcgtggtggtgtcgatggtagaacgaagcggcgtcgaagcctgtaaagcggcggtgcacaatcttctcgcgcaacgcgtcagtgggctgatcattaactatccgctggatgaccaggatgccattgctgtggaagctgcctgcactaatgttccggcgttatttcttgatgtctctgaccagacacccatcaacagtattattttctcccatgaggacggtacgcgactgggcgtggagcatctggtcgcattgggtcaccagcaaatcgcgctgttagcgggcccattaagttctgtctcggcgcgtctgcgtctggctggctggcataaatatctcactcgcaatcaaattcagccgatagcggaacgggaaggcgactggagtgccatgtccggttttcaacaaaccatgcaaatgctgaatgagggcatcgttcccactgcgatgctggttgccaacgatcagatggcgctgggcgcaatgcgcgccattaccgagtccgggctgcgcgttggtgcggatatctcggtagtgggatacgacgataccgaagatagctcatgttatatcccgccgttaaccaccatcaaacaggattttcgcctgctggggcaaaccagcgtggaccgcttgctgcaactctctcagggccaggcggtgaagggcaatcagctgttgcccgtctcactggtgaaaagaaaaaccaccctggcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcaggctgcaaacgacgaaaactacgctttagtagcttaataactctgatagtgctagtgtag +BCD12: + description: RBS - BiCistronic Design, medium strength (RBS part type, actually contains a small transcriptional unit and second RBS) + sequence: gggcccaagttcacttaaaaaggagatcaacaatgaaagcaattttcgtactgaaacatcttaatcatgctgcggagggtttct +B0032m: + description: RBS - Weiss RBS, medium strength. Modified from Bba_B0032 to adjust spacing in MC system + sequence: agagtcacacaggaaagtacta +BCD8: + description: RBS - BiCistronic Design, low strength (RBS part type, actually contains a small transcriptional unit and second RBS) + sequence: gggcccaagttcacttaaaaaggagatcaacaatgaaagcaattttcgtactgaaacatcttaatcatgcatcggaccgtttct +BCD2: + description: RBS - BiCistronic Design, high strength (RBS part type, actually contains a small transcriptional unit and second RBS) + sequence: gggcccaagttcacttaaaaaggagatcaacaatgaaagcaattttcgtactgaaacatcttaatcatgctaaggaggttttct +B0034m: + description: RBS - Weiss RBS, high strength. Modified from Bba_B0034 to adjust spacing in MC system. + sequence: agagaaagaggagaaatacta +B0033m: + description: RBS - Weiss RBS, low strength. Modified from Bba_B0033 to adjust spacing in MC system. + sequence: agagtcacacaggactacta +B0015: + description: Double terminator (B0010,B0012) + sequence: ccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttata +pBAD: + description: Controllable promoter - pBAD - araC regulated (pBAD promoter, inducible by expression of AraC, C0080) + seqeunce: acattgattatttgcacggcgtcacactttgctatgccatagcatttttatccataagattagcggatcctacctgacgctttttatcgcaactctctactgtttctccataccgtttttttgggctagc + +# +# Ecoflex +# + +eCFP(Ecoli): + description: placeholder + prefix: A + suffix: GGATCC + sequence: atggtgagcaagggcgaggagctgttcaccggggtggtgcccatcctggtcgagctggacggcgacgtgaacggccacaagttcagcgtgtccggcgagggcgagggcgatgccacctacggcaagctgaccctgaagttcatctgcaccaccggcaagctgcccgtgccctggcccaccctcgtgaccaccctgacctggggcgtgcagtgcttcagccgctaccccgaccacatgaagcagcacgacttcttcaagtccgccatgcccgaaggctacgtccaggagcgcaccatcttcttcaaggacgacggcaactacaagacccgcgccgaggtgaagttcgagggcgacaccctggtgaaccgcatcgagctgaagggcatcgacttcaaggaggacggcaacatcctggggcacaagctggagtacaactacatcagccacaacgtctatatcaccgccgacaagcagaagaacggcatcaaggccaacttcaagatccgccacaacatcgaggacggcagcgtgcagctcgccgaccactaccagcagaacacccccatcggcgacggccccgtgctgctgcccgacaaccactacctgagcacccagtccgccctgagcaaagaccccaacgagaagcgcgatcacatggtcctgctggagttcgtgaccgccgcc +eGFP(Ecoli): + description: placeholder + prefix: A + suffix: GGATCC + sequence: atgcgtaaaggagaagaacttttcactggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggagagggtgaaggtgatgcaacatacggaaaacttacccttaaatttatttgcactactggaaaactacctgttccatggccaacacttgtcactactttcggttatggtgttcaatgctttgcgagatacccagatcatatgaaacagcatgactttttcaagagtgccatgcccgaaggttatgtacaggaaagaactatatttttcaaagatgacgggaactacaagacacgtgctgaagtcaagtttgaaggtgatacccttgttaatagaatcgagttaaaaggtattgattttaaagaagatggaaacattcttggacacaaattggaatacaactataactcacacaatgtatacatcatggcagacaaacaaaagaatggaatcaaagttaacttcaaaattagacacaacattgaagatggaagcgttcaactagcagaccattatcaacaaaatactccaattggcgatggccctgtccttttaccagacaaccattacctgtccacacaatctgccctttcgaaagatcccaacgaaaagagagatcacatggtccttcttgagtttgtaacagctgctgggattacacatggcatggatgaactatacaaa +mCherry(Ecoli): + description: placeholder + prefix: A + suffix: GGATCC + sequence: atggtgagcaagggcgaggaggataacatggccatcatcaaggagttcatgcgcttcaaggtgcacatggagggctccgtgaacggccacgagttcgagatcgagggcgagggcgagggccgcccctacgagggcacccagaccgccaagctgaaggtgaccaagggtggccccctgcccttcgcctgggacatcctgtcccctcagttcatgtacggctccaaggcctacgtgaagcaccccgccgacatccccgactacttgaagctgtccttccccgagggcttcaagtgggagcgcgtgatgaacttcgaggacggcggcgtggtgaccgtgacccaggactcctccttgcaggacggcgagttcatctacaaggtgaagctgcgcggcaccaacttcccctccgacggccccgtaatgcagaagaagaccatgggctgggaggcctcctccgagcggatgtaccccgaggacggcgccctgaagggcgagatcaagcagaggctgaagctgaaggacggcggccactacgacgctgaggtcaagaccacctacaaggccaagaagcccgtgcagctgcccggcgcctacaacgtcaacatcaagttggacatcacctcccacaacgaggactacaccatcgtggaacagtacgaacgcgccgagggccgccactccaccggcggcatggacgagctgtacaag + +# +# CIDAR extension +# +J23101: + description: MoClo golden gate assembly AB part plasmid for J23101 (strength 1791x Anderson promoter; see http://parts.igem.org/Promoters/Catalog/Anderson) + sequence: TTTACAGCTAGCTCAGTCCTAGGTATTATGCTAGC +J23150: + description: MoClo golden gate assembly AB part for J23150 (strength ~400x Anderson promoter; see http://parts.igem.org/Promoters/Catalog/Anderson) + sequence: TGCTGTTCCGCTGGGCATGCTTTACGGCTAGCTCAGTCCTAGGTATTATGC +J23151: + description: MoClo golden gate assembly AB part for J23151 (strength ~900x Anderson promoter; see http://parts.igem.org/Promoters/Catalog/Anderson) + sequence: TGCTGTTCCGCTGGGCATGCTTGATGGCTAGCTCAGTCCTAGGTACAATGC +pOR1-OR2: + description: MoClo golden gate assembly AB part for pOR1-OR2 (lambda cI-repressable strong promoter) + sequence: TGCTGTTCCGCTGGGCATGCTGAGCTAACACCGTGCGTGTTGACAATTTTACCTCTGGCGGTGATAATGGTTGCAGC +Ptet: + description: MoClo golden gate assembly AB part for pLas-TetO (Promoter activated by LasR and repressed by TetR; encodes <3OC12 AHL AND aTc> logic). + sequence: CTAGCAAGGGTCCGGGTTCACCGAAATCTATCTCATTTGCTAGTTATAAAATTATGAAATTTGCGTAAATTCCCTATCAGTGATAGAGATTCAGAAGCAAT +pCin-TetO: + description: MoClo golden gate assembly AB part for pCin-TetO (Promoter activated by CinR and repressed by TetR; encodes <3OC14 AHL AND aTc> logic). + sequence: CGGAACTCTTGTGCGTGGCGCATGTCGTTATCGCAAAACCGCTGCACACTTTTGCGCGACATGCTCTGATCCCCCTCATCTGGGGGGGCCTATCTGAGGGAATTTACTTCCCTATCAGTGATAGAGATAATGAGCACATCCCTATCAGTGATAGAGATG +pCin-LacO: + description: MoClo golden gate assembly AB part for pCin-LacO (Promoter activated by CinR and repressed by LacI; encodes <3OC14 AHL AND IPTG> logic). + sequence: CGGAACTCTTGTGCGTGGCGCATGTCGTTATCGCAAAACCGCTGCACACTTTTGCGCGACATGCTCTGATCCCCCTCATCTGGGGGGGCCTATCTGAGGGAATTTACTTGTGAGCGCTCACAATTGATAATGAGCACAATTGTGAGCGGATAACAATTGCT +T7_promoter: + description: The T7 promoter binding site + sequence: TAATACGACTCACTATAG diff --git a/parts/parts/ecoli/origins.yaml b/parts/parts/ecoli/origins.yaml new file mode 100644 index 0000000..17a3808 --- /dev/null +++ b/parts/parts/ecoli/origins.yaml @@ -0,0 +1,52 @@ +# Origins +ColE1: + description: High copy (500-700 copies) common Escherichia coli origin + sequence: TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA +RSF1030: + description: High copy (>100 copies) Escherichia coli origin. Compatible with pUC/pBR322, p15A, ColA, CloDF13, and SC101 + sequence: AACGGAATAGCTGTTCGTTGACTTGATAGACCGATTGATTCATCATCTCATAAATAAAGAAAAACCACCGCTACCAACGGTGGTTTTCTCAAGGTTCGCTGAGCTACCAACTCTTTGAACCAAGGTAAGTGGGTTGGAGGACCGCACTCACCAAAATCTGTTCTTTCAGTTTAGCCTTAACAGGTGCATAACTTCAAGACAAAGTCCTCTAAATCAGTTACCAATGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCAACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGGCTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCTGCGGCGTTGGCTTCTTCCGGTGCTTTGCTTTTTGCTCACATGTTCTTTCCGGCTTTATCCCCTGATTCTGTGGATAACCGTATTACCGCTTTTGAGTGAGCTGACACCGCTCGCCGCAGTCGAACGACCGAGCGTAGCGAGTCAGTGAGCGAGGAAGCGGAAGA +pBR322: + description: Medium copy (~20) common Escherichia coli origin + sequence: CCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGTGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGTATACACTCCGCTATCGCTACGTGACTGGGTCATGGCTGCGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGGCAGCTGCGGTAAAGCTCATCAGCGTGGTCGTGCAGCGATTCACAGATGTCTGCCTGTTCATCCGCGTCCAGCTCGTTGAGTTTCTCCAGAAGCGTTAATGTCTGGCTTCTGATAAAGCGGGCCATGTTAAGGGCGGTTTTTTCCTGTTTGGTCACTGATGCCTCCGTGTAAGGGGGATTTCTGTTCATGGGGGTAATGATACCGATGAAACGAGAGAGGATGCTCACGATACGGGTTACTGATGATGAACATGCCCGGTTACTGGAACGTTGTGAGGGTAAACAACTGGCGGTATGGATGCGGCGGG +p15A: + description: Medium copy (10-12 copies) common Escherichia coli origin. pUC/pBR322, SC101, ColA, CloDF13, and RSF1030 + sequence: TTGAGATCGTTTTGGTCTGCGCGTAATCTCTTGCTCTGAAAACGAAAAAACCGCCTTGCAGGGCGGTTTTTCGAAGGTTCTCTGAGCTACCAACTCTTTGAACCGAGGTAACTGGCTTGGAGGAGCGCAGTCACCAAAACTTGTCCTTTCAGTTTAGCCTTAACCGGCGCATGACTTCAAGACTAACTCCTCTAAATCAATTACCAGTGGCTGCTGCCAGTGGTGCTTTTGCATGTCTTTCCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGACTGAACGGGGGGTTCGTGCATACAGTCCAGCTTGGAGCGAACTGCCTACCCGGAACTGAGTGTCAGGCGTGGAATGAGACAAACGCGGCCATAACAGCGGAATGACACCGGTAAACCGAAAGGCAGGAACAGGAGAGCGCACGAGGGAGCCGCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCACTGATTTGAGCGTCAGATTTCGTGATGCTTGTCAGGGGGGCGGAGCCTATGGAAA +ColE2: + description: Medium copy plasmid. Can be engineered easily for different copy number levels. Orthogonal to R6K origin + sequence: TTAAGGGGGGCGTACAACGGGAGTTATGGTAAATGGATCGGGTTTTCGGGAAGGATCGACAGGATTTGCCGTTGGGTGTAGTGTAAGCGACTGAAAAACAAACGCCCCGTAAATCGTGCTCTCACCGCCAAGATTGATCACGAAATTACAGGGCGCCGGGTTCCGCGTTTCCCGATGGGAAAGCGCGGTTAGTTAAACTGTGTACCGAGAGAAATCGTATCACATGAGCGCCGTACTTCAACGCTTCAGGGAAAAATTACCGCACAAACCGTACTGTACGAACGATTTCGCGTACGGCGTTCGCATTCTGCCGAAAAACATTGCCATTCTTGCCCGTTTCATCCAGCAGAACCAGCCACATGCACTGTACTGGCTTCCCTTTGACGTGGACCGGACGGGGGCATCAATCGACTGGAGCGACCGGAATTGTCCGGCCCCGAACATCACCGTAAAAAATCCCCGTAACGGGCACGCGCATCTGCTCTACGCGCTCGCCCTTCCTGTGAGAACTGCGCCGGATGCATCGGCTTCGGCGCTCAGATACGCTGCCGCTATTGAGCGTGCGTTGTGTGAAAAACTGGGCGCGGATGTGAATTACAGCGGCCTGATCTGCAAAAATCCGTGCCACCCTGAATGGCAGGAAGTGGAATGGCGCGAGGAACCCTACACTCTCGACGAACTGGCTGATTATCTCGATTTGAGCGCCTCAGCGCGCCGTAGCGTCGATAAAAATTACGGGCTGGGGCGAAACTACCATCTGTTCGAAAAGGTCCGTAAATGGGCCTACAGAGCGATTCGTCAGGGCTGGCCTGTATTCTCACAATGGCTTGATGCCGTGATCCAGCGTGTCGAAATGTACAACGCATCGCTTCCCGTTCCGCTTTCTCCGGCTGAATGTCGGGCTATTGGCAAGAGCATTGCGAAATATACACACAGGAAATTCTCACCAGAGGGATTTTCCGCTGTACAGGCCGCTCGAGGTCGCAAGGGCGGAACTAAATCTAAGCGCGCAGCAGTTCCTACATCAGCACGTTCGCTGAAGCCGTGGGAGGCATTAGGGATCAGTCGAGCGACGTACTACCGAAAATTAAAATGTGATCCAGATCTCGCAAAATAAGACCAGATAAGCCTTATCAGATAACAGCGCCCTTTTGGCGTCTTTTTGAGCAGTGATCTTTCTTGCGATTTTCTCCGTTCTGC +pWV01: + description: Medium copy broad host range plasmid. + sequence: CGATTTTTTATTAAAATTTTAGCGTTTATTTCGTTTAGTTATCGGCATAATCGTTAAAACAGGCGTTATCGTAGCGTAAAAGCCCTTGAGCGTAGCGTGGCTTTGCAGCGAAGATGTTGTCTGTTAGATTATGAAAGCCGATGACTGAATGAAATAATAAGCGCAGCGCCCTTCTATTTCGGTTGGAGGAGGCTCAAGGGAGTATGAGGGAATGAAATTCCCTCATGGGTTTGATTTTAAAAATTGCTTGCAATTTTGCCGAGCGGTAGCGCTGGAAAATTTTTGAAAAAAATTTGGAATTTGGAAAAAAATGGGGGGAAAGGAAGCGAATTTTGCTTCCGTACTACGACCCCCCATTAAGTGCCGAGTGCCAATTTTTGTGCCAAAAACGCTCTATCCCAACTGGCTCAAGGGTTTAAGGGGTTTTTCAATCGCCAACGAATCGCCAACGTTTTCGCCAACGTTTTTTATAAATCTATATTTAAGTAGCTTTATTGTTGTTTTTATGATTACAAAGTGATACACTAACTTTATAAAATTATTTGATTGGAGTTTTTTAAATGGTGATTTCAGAATCGAAAAAAAGAGTTATGATTTCTCTGACAAAAGAGCAAGATAAAAAATTAACAGATATGGCGAAACAAAAAGGTTTTTCAAAATCTGCGGTTGCGGCGTTAGCTATAGAAGAATATGCAAGAAAGGAATCAGAACAAAAAAAATAAGCGAAAGCTCGCGTTTTTAGAAGGATACGAGTTTTCGCTACTTGTTTTTGATAAGGTAATTATATCATGGCTATTAAAAATACTAAAGCTAGAAATTTTGGATTTTTATTATATCCTGACTCAATTCCTAATGATTGGAAAGAAAAATTAGAGAGTTTGGGCGTATCTATGGCTGTCAGTCCTTTACACGATATGGACGAAAAAAAAGATAAAGATACATGGAATAGTAGTGATGTTATACGAAATGGAAAGCACTATAAAAAACCACACTATCACGTTATATATATTGCACGAAATCCTGTAACAATAGAAAGCGTTAGGAACAAGATTAAGCGAAAATTGGGGAATAGTTCAGTTGCTCATGTTGAGATACTTGATTATATCAAAGGTTCATATGAATATTTGACTCATGAATCAAAGGACGCTATTGCTAAGAATAAACATATATACGACAAAAAAGATATTTTGAACATTAATGATTTTGATATTGACCGCTATATAACACTTGATGAAAGCCAAAAAAGAGAATTGAAGAATTTACTTTTAGATATAGTGGATGACTATAATTTGGTAAATACAAAAGATTTAATGGCTTTTATTCGCCTTAGGGGAGCGGAGTTTGGAATTTTAAATACGAATGATGTAAAAGATATTGTTTCAACAAACTCTAGCGCCTTTAGATTATGGTTTGAGGGCAATTATCAGTGTGGATATAGAGCAAGTTATGCAAAGGTTCTTGATGCTGAAACGGGGGAAATAAAATGACAAACAAAGAAAAAGAGTTATTTGCTGAAAATGAGGAATTAAAAAAAGAAATTAAGGACTTAAAAGAGCGTATTGAAAGATACAGAGAAATGGAAGTTGAATTAAGTACAACAATAGATTTATTGAGAGGAGGGATTATTGAATAAATAAAAGCCCCCTGACGAAAGTCGAAGGGGGTTTTTATTTTGGTTTGATGTTGCGATTAATAGCAATACAATTGCAATAAACAAAATGATC +pBAV1KT5: + description: High copy (~350 E.coli, ~70 Bacillus subtilis, ~3 Agrobacterium tumefaciens, ~60 Acinetobacter baylyi, 100-500 Streptococcus pneumoniae) broad host range vector derived from pWV01. + description: CCAAGCACTAGGCGATTTTTTATTAAAATTTTAGCGTTTATTTCGTTTAGTTATCGGCATAATCGTTAAAACAGGCGTTATCGTAGCGTAAAAGCCCTTGAGCGTAGCGTGGCTTTGCAGCGAAGATGTTGTCTGTTAGATTATGAAAGCCGATGACTGAATGAAATAATAAGCGCAGCGCCCTTCTATTTCGGTTGGAGGAGGCTCAAGGGAGTATGAGGGAATGAAATTCCCTCATGGGTTTGATTTTAAAAATTGCTTGCAATTTTGCCGAGCGGTAGCGCTGGAAAATTTTTGAAAAAAATCTGGAATTTGGAAAAAAATGGGGGGAAAGGAAGCGAATTTTGCTTCCGTACTACGACCCCCCATTAAGTGCCGAGTGCCAATTTTTGTGCCAAAAACGCTCTATCCCAACTGGCTCAAGGGTTTAAGGGGTTTTTCAATCGCCAACGAATCGCCAACGTTTTCGCCAACGTTTTTTATAAATCTATATTTAAGTAGCTTTATTGTTGTTTTTATGATTACAAAGTGATACACTAACTTTATAAAATTATTTGATTGGAGTTTTTTAAATGGTGATTTCAGAATCGAAAAAAAGAGTTATGATTTCTCTGACAAAAGAGCAAGATAAAAAATTAACAGATATGGCGAAACAAAAAGGTTTTTCAAAATCTGCGGTTGCGGCGTTAGCTATAGAAGAATATGCAAGAAAGGAATCAGAACAAAAAAAATAAGCGAAAGCTCGCGTTTTTAGAAGGATACGAGTTTTCGCTACTTGTTTTTGATAAGGTAATTATATCATGGCTATTAAAAATACTAAAGCTAGAAATTTTGGATTTTTATTATATCCTGACTCAATTCCTAATGATTGGAAGGAAAAATTAGAGAGTTTGGGCGTATCTATGGCTGTCAGTCCTTTACACGATATGGACGAAAAAAAAGATAAAGATACATGGAATAGTAGTGATGTTATACGAAATGGAAAGCACTATAAAAAACCACACTATCACGTTATATATATTGCACGAAATCCTGTAACAATAGAAAGCGTTAGGAACAAGATTAAGCGAAAATTGGGGAATAGTTCAGTTGCTCATGTTGAGATACTTGATTATATCAAAGGTTCATATGAATATTTGACTCATGAATCAAAGGACGCTATTGCTAAGAATAAACATATATACGACAAAAAAGATATTTTGAACATTAATGATTTTGATATTGACCGCTATATAACACTTGATGAAAGCCAAAAAAGAGAATTGAAGAATTTACTCTTAGATATAGTGGATGACTATAATTTGGTAAATACAAAAGATTTAATGGCTTTTATTCGCCTTAGGGGAGCGGAGTTTGGAATTTTAAATACGAATGATGTAAAAGATATTGTTTCAACAAACTCTAGCGCCTTTAGATTATGGTTTGAGGGCAATTATCAGTGTGGATATAGAGCAAGTTATGCAAAGGTTCTTGATGCTGAAACGGGGGAAATAAAATGACACTAGGTCTAGGGCGGCGGATTTGTCCTACTCAGGAGAGCGTTCACCGACAAACAACAGATAAAACGAAAGGCCCAGTCTTTCGACTGAGCCTTTCGTTTTATTTG +RSF1010: + description: Low-med copy very broad host range origin (most gram-negative and some gram-positive) + sequence: GTCCTGGCCGCCACATGAGGCATGTTCCGCGTCCTTGCAATACTGTGTTTACATACAGTCTATCGCTTAGCGGAAAGTTCTTTTACCCTCAGCCGAAATGCCTGCCGTTGCTAGACATTGCCAGCCAGTGCCCGTCACTCCCGTACTAACTGTCACGAACCCCTGCAATAACTGTCACGCCCCCCTGCAATAACTGTCACGAACCCCTGCAATAACTGTCACGCCCCCAAACCTGCAAACCCAGCAGGGGCGGGGGCTGGCGGGGTGTTGGAAAAATCCATCCATGATTATCTAAGAATAATCCACTAGGCGCGGTTATCAGCGCCCTTGTGGGGCGCTGCTGCCCTTGCCCAATATGCCCGGCCAGAGGCCGGATAGCTGGTCTATTCGCTGCGCTAGGCTACACACCGCCCCACCGCTGCGCGGCAGGGGGAAAGGCGGGCAAAGCCCGCTAAACCCCACACCAAACCCCGCAGAAATACGCTGGAGCGCTTTTAGCCGCTTTAGCGGCCTTTCCCCCTACCCGAAGGGTGGGGGCGCGTGTGCAGCCCCGCAGGGCCTGTCTCGGTCGATCATTCAGCCCGGCTCATCCTTCTGGCGTGGCGGCAGACCGAACAAGGCGCGGTCGTGGTCGCGTTCAAGGTACGCATCCATTGCCGCCATGAGCCGATCCTCCGGCCACTCGCTGCTGTTCACCTTGGCCAAAATCATGGCCCCCACCAGCACCTTGCGCCTTGTTTCGTTCTTGCGCTCTTGCTGCTGTTCCCTTGCCCGCACCCGCTGAATTTCGGCATTGATTCGCGCTCGTTGTTCTTCGAGCTTGGCCAGCCGATCCGCCGCCTTGTTGCTCCCCTTAACCATCTTGACACCCCATTGTTAATGTGCTGTCTCGTAGGCTATCATGGAGGCACAGCGGCGGCAATCCCGACCCTACTTTGTAGGGGAGGGCGCACTTACCGGTTTCTCTTCGAGAAACTGGCCTAACGGCCACCCTTCGGGCGGTGCGCTCTCCGAGGGCCATTGCATGGAGCCGAAAAGCAAAAGCAACAGCGAGGCAGCATGGCGATTTATCACCTTACGGCGAAAACCGGCAGCAGGTCGGGCGGCCAATCGGCCAGGGCCAAGGCCGACTACATCCAGCGCGAAGGCAAGTATGCCCGCGACATGGATGAAGTCTTGCACGCCGAATCCGGGCACATGCCGGAGTTCGTCGAGCGGCCCGCCGACTACTGGGATGCTGCCGACCTGTATGAACGCGCCAATGGGCGGCTGTTCAAGGAGGTCGAATTTGCCCTGCCGGTCGAGCTGACCCTCGACCAGCAGAAGGCGCTGGCGTCCGAGTTCGCCCAGCACCTGACCGGTGCCGAGCGCCTGCCGTATACGCTGGCCATCCATGCCGGTGGCGGCGAGAACCCGCACTGCCACCTGATGATCTCCGAGCGGATCAATGACGGCATCGAGCGGCCCGCCGCTCAGTGGTTCAAGCGGTACAACGGCAAGACCCCGGAGAAGGGCGGGGCACAAAAGACCGAAGCGCTCAAGCCCAAGGCATGGCTTGAGCAGACCCGCGAGGCATGGGCCGACCATGCCAACCGGGCATTAGAGCGGGCTGGCCACGACGCCCGCATTGACCACAGAACACTTGAGGCGCAGGGCATCGAGCGCCTGCCCGGTGTTCACCTGGGGCCGAACGTGGTGGAGATGGAAGGCCGGGGCATCCGCACCGACCGGGCAGACGTGGCCCTGAACATCGACACCGCCAACGCCCAGATCATCGACTTACAGGAATACCGGGAGGCAATAGACCATGAACGCAATCGACAGAGTGAAGAAATCCAGAGGCATCAACGAGTTAGCGGAGCAGATCGAACCGCTGGCCCAGAGCATGGCGACACTGGCCGACGAAGCCCGGCAGGTCATGAGCCAGACCCAGCAGGCCAGCGAGGCGCAGGCGGCGGAGTGGCTGAAAGCCCAGCGCCAGACAGGGGCGGCATGGGTGGAGCTGGCCAAAGAGTTGCGGGAGGTAGCCGCCGAGGTGAGCAGCGCCGCGCAGAGCGCCCGGAGCGCGTCGCGGGGGTGGCACTGGAAGCTATGGCTAACCGTGATGCTGGCTTCCATGATGCCTACGGTGGTGCTGCTGATCGCATCGTTGCTCTTGCTCGACCTGACGCCACTGACAACCGAGGACGGCTCGATCTGGCTGCGCTTGGTGGCCCGATGAAGAACGACAGGACTTTGCAGGCCATAGGCCGACAGCTCAAGGCCATGGGCTGTGAGCGCTTCGATATCGGCGTCAGGGACGCCACCACCGGCCAGATGATGAACCGGGAATGGTCAGCCGCCGAAGTGCTCCAGAACACGCCATGGCTCAAGCGGATGAATGCCCAGGGCAATGACGTGTATATCAGGCCCGCCGAGCAGGAGCGGCATGGTCTGGTGCTGGTGGACGACCTCAGCGAGTTTGACCTGGATGACATGAAAGCCGAGGGCCGGGAGCCTGCCCTGGTAGTGGAAACCAGCCCGAAGAACTATCAGGCATGGGTCAAGGTGGCCGACGCCGCAGGCGGTGAACTTCGGGGGCAGATTGCCCGGACGCTGGCCAGCGAGTACGACGCCGACCCGGCCAGCGCCGACAGCCGCCACTATGGCCGCTTGGCGGGCTTCACCAACCGCAAGGACAAGCACACCACCCGCGCCGGTTATCAGCCGTGGGTGCTGCTGCGTGAATCCAAGGGCAAGACCGCCACCGCTGGCCCGGCGCTGGTGCAGCAGGCTGGCCAGCAGATCGAGCAGGCCCAGCGGCAGCAGGAGAAGGCCCGCAGGCTGGCCAGCCTCGAACTGCCCGAGCGGCAGCTTAGCCGCCACCGGCGCACGGCGCTGGACGAGTACCGCAGCGAGATGGCCGGGCTGGTCAAGCGCTTCGGTGATGACCTCAGCAAGTGCGACTTTATCGCCGCGCAGAAGCTGGCCAGCCGGGGCCGCAGTGCCGAGGAAATCGGCAAGGCCATGGCCGAGGCCAGCCCAGCGCTGGCAGAGCGCAAGCCCGGCCACGAAGCGGATTACATCGAGCGCACCGTCAGCAAGGTCATGGGTCTGCCCAGCGTCCAGCTTGCGCGGGCCGAGCTGGCACGGGCACCGGCACCCCGCCAGCGAGGCATGGACAGGGGCGGGCCAGATTTCAGCATGTAGTGCTTGCGTTGGTACTCACGCCTGTTATACTATGAGTACTCACGCACAGAAGGGGGTTTTATGGAATACGAAAAAAGCGCTTCAGGGTCGGTCTACCTGATCAAAAGTGACAAGGGCTATTGGTTGCCCGGTGGCTTTGGTTATACGTCAAACAAGGCCGAGGCTGGCCGCTTTTCAGTCGCTGATATGGCCAGCCTTAACCTTGACGGCTGCACCTTGTCCTTGTTCCGCGAGGACAAGCCTTTCGGCCCCGGCAAGTTTCTCGGTGACTGATATGAAAGACCAAAAGGACAAGCAGACCGGCGACCTGCTGGCCAGCCCTGACGCTGTACGCCAAGCGCGATATGCCGAGCGCATGAAGGCCAAAGGGATGCGTCAGCGCAAGTTCTGGCTGACCGACGACGAATACGAGGCGCTGCGCGAGTGCCTGGAAGAACTCAGAGCGGCGCAGGGCGGGGGTAGTGACCCCGCCAGCGCCTAACCACCAACTGCCTGCAAAGGAGGCAATCAATGGCTACCCATAAGCCTATCAATATTCTGGAGGCGTTCGCAGCAGCGCCGCCACCGCTGGACTACGTTTTGCCCAACATGGTGGCCGGTACGGTCGGGGCGCTGGTGTCGCCCGGTGGTGCCGGTAAATCCATGCTGGCCCTGCAACTGGCCGCACAGATTGCAGGCGGGCCGGATCTGCTGGAGGTGGGCGAACTGCCCACCGGCCCGGTGATCTACCTGCCCGCCGAGGACCCGCCCACCGCCATTCATCACCGCCTGCACGCCCTTGGGGCGCACCTCAGCGCCGAGGAACGGCAAGCCGTGGCTGACGGCCTGCTGATCCAGCCGCTGATCGGCAGCCTGCCCAACATCATGGCCCCGGAGTGGTTCGACGGCCTCAAGCGCGCCGCCGAGGGCCGCCGCCTGATGGTGCTGGACACGCTGCGCCGGTTCCACATCGAGGAAGAAAACGCCAGCGGCCCCATGGCCCAGGTCATCGGTCGCATGGAGGCGATCGCCGCCGATACCGGGTGCTCTATCGTGTTCCTGCACCATGCCAGCAAGGGCGCGGCCATGATGGGCGCAGGCGACCAGCAGCAGGCCAGCCGGGGCAGCTCGGTACTGGTCGATAACATCCGCTGGCAGTCCTACCTGTCGAGCATGACCAGCGCCGAGGCCGAGGAATGGGGTGTGGACGACGACCAGCGCCGGTTCTTCGTCCGCTTCGGTGTGAGCAAGGCCAACTATGGCGCACCGTTCGCTGATCGGTGGTTCAGGCGGCATGACGGCGGGGTGCTCAAGCCCGCCGTGCTGGAGAGGCAGCGCAAGAGCAAGGGGGTGCCCCGTGGTGAAGCCTAAGAACAAGCACAGCCTCAGCCACGTCCGGCACGACCCGGCGCACTGTCTGGCCCCCGGCCTGTTCCGTGCCCTCAAGCGGGGCGAGCGCAAGCGCAGCAAGCTGGACGTGACGTATGACTACGGCGACGGCAAGCGGATCGAGTTCAGCGGCCCGGAGCCGCTGGGCGCTGATGATCTGCGCATCCTGCAAGGGCTGGTGGCCATGGCTGGGCCTAATGGCCTAGTGCTTGGCCCGGAACCCAAGACCGAAGGCGGACGGCAGCTCCGGCTGTTCCTGGAACCCAAGTGGGAGGCCGTCACCGCTGAATGCCATGTGGTCAAAGGTAGCTATCGGGCGCTGGCAAAGGAAATCGGGGCAGAGGTCGATAGTGGTGGGGCGCTCAAGCACATACAGGACTGCATCGAGCGCCTTTGGAAGGTATCCATTATCGCCCAGAATGGCCGCAAGCGGCAGGGGTTTCGGCTGCTGTCGGAGTACGCCAGCGACGAGGCGGACGGGCGCCTGTACGTGGCCCTGAACCCCTTGATCGCGCAGGCCGTCATGGGTGGCGGCCAGCATGTGCGCATCAGCATGGACGAGGTGCGGGCGCTGGACAGCGAAACCGCCCGCCTGCTGCACCAGCGGCTGTGTGGCTGGATCGACCCCGGCAAAACCGGCAAGGCTTCCATAGATACCTTGTGCGGCTATGTCTGGCCGTCAGAGGCCAGTGGTTCGACCATGCGCAAGCGCCGCCAGCGGGTGCGCGAGGCGTTGCCGGAGCTGGTCGCGCTGGGCTGGACGGTAACCGAGTTCGCGGCGGGCAAGTACGACATCACCCGGCCCAAGGCGGCAGGCTGACCCCCCCCACTCTATTGTAAACAAGACATTTTTATCTTTTATATTCAATGGCTTATTTTCCTGCTAATTGGTAATACCATGAAAAATACCATGCTCAGAAAAGGCTTAACAATATTTTGAAAAATTGCCTACTGAGCGCTGCCGCACAGCTCCATAGGCCGCTTTCCTGGCTTTGCTTCCAGATGTATGCTCTTATGCTCC +SC101: + description: Low copy (~5 copies) Escherichia coli origin. Compatible with pUC/pBR322, p15A, ColA, CloDF13, and RSF1030 + sequence: ATGCAGCGATTTGCCCGAGCTTGCGAGGGTGCTACTTAAGCCTTTAGGGTTTTAAGGTCTGTTTTGTAGAGGAGCAAACAGCGTTTGCGACATCCTTTTGTAATACTGCGGAACTGACTAAAGTAGTGAGTTATACACAGGGCTGGGATCTATTCTTTTTATCTTTTTTTATTCTTTCTTTATTCTATAAATTATAACCACTTGAATATAAACAAAAAAAACACACAAAGGTCTAGCGGAATTTACAGAGGGTCTAGCAGAATTTACAAGTTTTCCAGCAAAGGTCTAGCAGAATTTACAGATACCCACAACTCAAAGGAAAAGGACTAGTAATTATCATTGACTAGCCCATCTCAATTGGTATAGTGATTAAAATCACCTAGACCAATTGAGATGTATGTCTGAATTAGTTGTTTTCAAAGCAAATGAACTAGCGACTAGTCGCTATGACTTAACGGAGCATGAAACCAAGCTAATTTTATGCTGTGTGGCACTACTCAACCCCACGATTGAAAACCCTACAAGGAAAGAACGGACGGTATCGTTCACTTATAACCAATACGTTCAGATGATGAACATCAGTAGGGAAAATGCTTATGGTGTATTAGCTAAAGCAACCAGAGAGCTGATGACGAGAACTGTGGAAATCAGGAATCCTTTGGTTAAAGGCTTTGAGATTTTCCAGTGGACAAACTATGCCAAGTTCTCAAGCGAAAAATTAGAATTAGTTTTTAGTGAAGAGATATTGCCTTATCTTTTCCAGTTAAAAAAATTCATAAAATATAATCTGGAACATGTTAAGTCTTTTGAAAACAAATACTCTATGAGGATTTATGAGTGGTTATTAAAAGAACTAACACAAAAGAAAACTCACAAGGCAAATATAGAGATTAGCCTTGATGAATTTAAGTTCATGTTAATGCTTGAAAATAACTACCATGAGTTTAAAAGGCTTAACCAATGGGTTTTGAAACCAATAAGTAAAGATTTAAACACTTACAGCAATATGAAATTGGTGGTTGATAAGCGAGGCCGCCCGACTGATACGTTGATTTTCCAAGTTGAACTAGATAGACAAATGGATCTCGTAACCGAACTTGAGAACAACCAGATAAAAATGAATGGTGACAAAATACCAACAACCATTACATCAGATTCCTACCTACATAACGGACTAAGAAAAACACTACACGATGCTTTAACTGCAAAAATTCAGCTCACCAGTTTTGAGGCAAAATTTTTGAGTGACATGCAAAGTAAGTATGATCTCAATGGTTCGTTCTCATGGCTCACGCAAAAACAACGAACCACACTAGAGAACATACTGGCTAAATACGGAAGGATCTGAGGTTCTTATGGCTCTTGTATCTATCAGTGAAGCATCAAGACTAACAAACAAAAGTAGAACAACTGTTCACCGTTACATATCAAAGGGAAAACTGTCCATATGCACAGATGAAAACGGTGTAAAAAAGATAGATACATCAGAGCTTTTACGAGTTTTTGGTGCA +pRK2: + description: Low copy (4-7 copies) origin with broad gram-negative host range + sequence: GTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGGGGGATCAGGACCGCTGCCGGAGCGCAACCCACTCACTACAGCAGAGCCATGTAGGGCCGCCGGCGTTGTGGATACCTCGCGGAAAACTTGGCCCTCACTGACAGATGAGGGGCGGACGTTGACACTTGAGGGGCCGACTCACCCGGCGCGGCGTTGACAGATGAGGGGCAGGCTCGATTTCGGCCGGCGACGTGGAGCTGGCCAGCCTCGCAAATCGGCGAAAACGCCTGATTTTACGCGAGTTTCCCACAGATGATGTGGACAAGCCTGGGGATAAGTGCCCTGCGGTATTGACACTTGAGGGGCGCGACTACTGACAGATGAGGGGCGCGATCCTTGACACTTGAGGGGCAGAGTGCTGACAGATGAGGGGCGCACCTATTGACATTTGAGGGGCTGTCCACAGGCAGAAAATCCAGCATTTGCAAGGGTTTCCGCCCGTTTTTCGGCCACCGCTAACCTGTCTTTTAACCTGCTTTTAAACCAATATTTATAAACCTTGTTTTTAACCAGGGCTGCGCCCTGTGCGCGTGACCGCGCACGCCGAAGGGGGGTGCCCCCCCTTCTCGAACCCTCCCGGCCCGCTAACGCGGGCCTCCCATCCCCCCAGGGGCTGCGCCCCTCGGCCGCGAACGGCCTCACCCCAAAAATGGCAGCCACGTAGAAAGCCAGTCCGCAGAAACGGTGCTGACCCCGGATGAATGTCAGCTACTGGGCTATCTGGACAAGGGAAAACGCAAGCGCAAAGAGAAAGCAGGTAGCTTGCAGTGGGCTTACATGGCGATAGCTAGACTGGGCGGTTTTATGGACAGCAAGCGAACCGGAATTGCCAGCTGGGGCGCCCTCTGGTAAGGTTGGGAAGCCCTGCAAAGTAAACTGGATGGCTTTCTTGCCGCCAAGGATCTGATGGCGCAGGGGATCAAGATCGACGGATCGATCCGGGGAATTAATTCCGGGGCAATCCCGCAAGGAGGGTGAATGAATCGGACGTTTGACCGGAAGGCATACAGGCAAGAACTGATCGACGCGGGGTTTTCCGCCGAGGATGCCGAAACCATTGCAAGCCGCACCGTCATGCGTGCGCCCCGCGAAACCTTCCAGTCCGTCGGCTCGATAGTCCAGCAAGCTACGGCCAAGATCGAGCGCGACAGCGTGCAACTGGCTCCCCCTGCCCTGCCCGCGCCATCGGCCGCCGTGGAGCGTTCGCGTCGCCTCGAACAGGAGGCGGCAGGTTTGGCGAAGTCGATGACCATCGACACGCGAGGAACTATGACGACCAAGAAGCGAAAAACCGCCGGCGAGGACCTGGCAAAACAGGTCAGCGAAGCCAAGCAGGCCGCGTTGCTGAAACACACGAAGCAGCAGATCAAGGAAATGCAGCTTTCCTTGTTCGATATTGCGCCGTGGCCGGACACGATGCGAGCCATGCCAAACGACACGGCCCGCTCTGCCCTGTTCACCACGCGCAACAAGAAAATCCCGCGCGAGGCGCTGCAAAACAAGGTCATTTTCCACGTCAACAAGGACGTGAAGATCACCTACACCGGCGTCGAGCTGCGGGCCGACGATGACGAACTGGTGTGGCAGCAAGTGTTGGAGTACGCGAAGCGCACCCCTATCGGCGAGCCGATCACCTTCACGTTCTACGAGCTTTGCCAGGACCTGGGCTGGTCGATCAATGGCCGGTATTACACGAAGGCCGAGGAATGCCTGTCGCGCCTACAGGCGACGGCCATGGGCTTCACGTCCGACCGCGTTGGGCACCTGGAATCGGTGTCGCTGCTGCACCGCTTCCGCGTCCTGGACCGTGGCAAGAAAACGTCCCGTTGCCAGGTCCTGATCGACGAGGAAATCGTCGTGCTGTTTGCTGGCGACCACTACACGAAATTCATATGGGAGAAGTACCGCAAGCTGTCGCCGACGGCCCGACGGATGTTCGACTATTTCAGCTCGCACCGGGAGCCGTACCCGCTCAAGCTGGAAACCTTCCGCCTCATGTGCGGATCGGATTCCACCCGCGTGAAGAAGTGGCGCGAGCAGGTCGGCGAAGCCTGCGAAGAGTTGCGAGGCAGCGGCCTGGTGGAACACGCCTGGGTCAATGATGACCTGGTGCATTGCAAACGCTAGGGCCTTGTGGGGTCAGTTCCGGCTGGGGGTTCAGCAGCCACCTG +pRO1600: + description: Broad host range plasmid (varies per host) for gram negative cells. + sequence: GCCGCTGGTGCCGCTGGTTGGACGCCAAGGGTGAATCCGCCTCGATACCCTGATTACTCGCTTCCTGCGCCCTCTCAGGCGGCGATAGGGGACTGGTAAAACGGGGATTGCCCAGACGCCTCCCCCGCCCCTTCAGGGGCACAAATGCGGCCCCAACGGGGCCACGTAGTGGTGCGTTTTTTGCGTTTCCACCCTTTTCTTCCTTTTCCCTTTTAAACCTTTTAGGACGTCTACAGGCCACGTAATCCGTGGCCTGTAGAGTTTAAAAAGGGACGGATTTGTTGCCATTAAGGGACGGATTTGTTGTTAAGAAGGGACGGATTTGTTGTTGTAAAGGGACGGATTTGTTGTATTGTGGGACGCAGATACAGTGTCCCCTTATACACAAGGAATGTCGAACGTGGCCTCACCCCCAATGGTTTACAAAAGCAATGCCCTGGTCGAGGCCGCGTATCGCCTCAGTGTTCAGGAACAGCGGATCGTTCTGGCCTGTATTAGCCAGGTGAAGAGGAGCGAGCCTGTCACCGATGAAGTGATGTATTCAGTGACGGCGGAGGACATAGCGACGATGGCGGGTGTCCCTATCGAATCTTCCTACAACCAGCTCAAAGAAGCGGCCCTGCGCCTGAAACGGCGGGAAGTCCGGTTAACCCAAGAGCCCAATGGCAAGGGGAAAAGACCGAGTGTGATGATTACCGGCTGGGTGCAAACAATCATCTACCGGGAGGGTGAGGGCCGTGTAGAACTCAGGTTCACCAAAGACATGCTGCCGTACCTGACGGAACTCACCAAACAGTTCACCAAATACGCCTTGGCTGACGTGGCCAAGATGGACAGCACCCACGCGATCAGGCTTTACGAGCTGCTCATGCAATGGGACAGCATCGGCCAGCGCGAAATAGAAATTGACCAGCTGCGAAAGTGGTTTCAACTGGAAGGCCGGTATCCCTCGATCAAGGACTTCAAGTTGCGAGTGCTTGATCCAGCCGTGACGCAGATCAACGAGCACAGCCCGCTACAGGTGGAGTGGGCGCAGCGAAAGACCGGGCGCAAGGTCACACATCTGTTGTTCAGTTTTGGACCGAAGAAGCCCGCCAAGGCGGTGGGTAAGGCCCCAGCGAAGCGCAAGGCCGGGAAGATTTCAGATGCTGAGATCGCGAAACAGGCTCGCCCTGGTGAGACATGGGAAGCGGCCCGCGCTCGACTAACCCAGATGCCGCTGGATCTGGCCTAGAGGCCGTGGCCACCACGGCCCGGCCTGCCTTTC +ColA: + description: Medium copy (20-40 copies) Escherichia coli origin. Compatible with pUC/pBR322, p15A, RSF1030, CloDF13, and SC101 + sequence: TGGTGTCGGGAATCCGTAAAGGATCTTCTTGAGATCCTTTTACGATCGTCGTAATCTCCTGCTCTGTAAACGAAAAAACCGCCTGGGGAGGCGGTTTGATCGAAGGTTAAGTCAGTTGGGGAACTGCTTAACCTGGTAACTGGCTTTAGTGGAGCGCAGATACCAAATACTGTCCTTTCAGTGTAGCCTCTGTTAGGCCACCACTTCAAGACTCTCGATATCTAAATCCACTAATTCTCAGTTACCAATGGCTGCTGCCAGTGGCGTTTTGTCGTGTCTTTCCGGGTTGGACTCAAGATGATAGTTACCGGATAAGGCGCAGCAGTCGGGCTGAACGGGGGGTTCTTGCACACAGCCCAGCTTGGAGCGAACTGTCTACACGGAACGGGACGTGGTGATTTGGGTAAAGCCTCCACCACAACACGGACGCCGCAGGACGGGAACAGGAGAGCGCAAGAGGGAGCCATCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCACTGATTTGAGCGTCAGATTTCGTGATGTTCGTCAGGGGGGCGGAGCCTATGGAAAAACGGCTTCGCTCCGGCCTTATTGTCTCTCTGCTAAGTATCCTCCTGGCATCTTCTAGGACGTTT +BAC: + description: Single copy origin built to create Bacterial Artificial Chromosomes, which allow for cloning and stable maintenance of very large DNA fragments (up to 300kb) + sequence: CATGGAGCGGCGTAACCGTCGCACAGGAAGGACAGAGAAAGCGCGGATCTGGGAAGTGACGGACAGAACGGTCAGGACCTGGATTGGGGAGGCGGTTGCCGCCGCTGCTGCTGACGGTGTGACGTTCTCTGTTCCGGTCACACCACATACGTTCCGCCATTCCTATGCGATGCACATGCTGTATGCCGGTATACCGCTGAAAGTTCTGCAAAGCCTGATGGGACATAAGTCCATCAGTTCAACGGAAGTCTACACGAAGGTTTTTGCGCTGGATGTGGCTGCCCGGCACCGGGTGCAGTTTGCGATGCCGGAGTCTGATGCGGTTGCGATGCTGAAACAATTATCCTGAGAATAAATGCCTTGGCCTTTATATGGAAATGTGGAACTGAGTGGATATGCTGTTTTTGTCTGTTAAACAGAGAAGCTGGCTGTTATCCACTGAGAAGCGAACGAAACAGTCGGGAAAATCTCCCATTATCGTAGAGATCCGCATTATTAATCTCAGGAGCCTGTGTAGCGTTTATAGGAAGTAGTGTTCTGTCATGATGCCTGCAAGCGGTAACGAAAACGATTTGAATATGCCTTCAGGAACAATAGAAATCTTCGTGCGGTGTTACGTTGAAGTGGAGCGGATTATGTCAGCAATGGACAGAACAACCTAATGAACACAGAACCATGATGTGGTCTGTCCTTTTACAGCCAGTAGTGCTCGCCGCAGTCGAGCGACAGGGCGAAGCCCTCGAGTGAGCGAGGAAGCACCAGGGAACAGCACTTATATATTCTGCTTACACACGATGCCTGAAAAAACTTCCCTTGGGGTTATCCACTTATCCACGGGGATATTTTTATAATTATTTTTTTTATAGTTTTTAGATCTTCTTTTTTAGAGCGCCTTGTAGGCCTTTATCCATGCTGGTTCTAGAGAAGGTGTTGTGACAAATTGCCCTTTCAGTGTGACAAATCACCCTCAAATGACAGTCCTGTCTGTGACAAATTGCCCTTAACCCTGTGACAAATTGCCCTCAGAAGAAGCTGTTTTTTCACAAAGTTATCCCTGCTTATTGACTCTTTTTTATTTAGTGTGACAATCTAAAAACTTGTCACACTTCACATGGATCTGTCATGGCGGAAACAGCGGTTATCAATCACAAGAAACGTAAAAATAGCCCGCGAATCGTCCAGTCAAACGACCTCACTGAGGCGGCATATAGTCTCTCCCGGGATCAAAAACGTATGCTGTATCTGTTCGTTGACCAGATCAGAAAATCTGATGGCACCCTACAGGAACATGACGGTATCTGCGAGATCCATGTTGCTAAATATGCTGAAATATTCGGATTGACCTCTGCGGAAGCCAGTAAGGATATACGGCAGGCATTGAAGAGTTTCGCGGGGAAGGAAGTGGTTTTTTATCGCCCTGAAGAGGATGCCGGCGATGAAAAAGGCTATGAATCTTTTCCTTGGTTTATCAAACGTGCGCACAGTCCATCCAGAGGGCTTTACAGTGTACATATCAACCCATATCTCATTCCCTTCTTTATCGGGTTACAGAACCGGTTTACGCAGTTTCGGCTTAGTGAAACAAAAGAAATCACCAATCCGTATGCCATGCGTTTATACGAATCCCTGTGTCAGTATCGTAAGCCGGATGGCTCAGGCATCGTGTCTCTGAAAATCGACTGGATCATAGAGCGTTACCAGCTGCCTCAAAGTTACCAGCGTATGCCTGACTTCCGCCGCCGCTTCCTGCAGGTCTGTGTTAATGAGATCAACAGCAGAACTCCAATGCGCCTCTCATACATTGAGAAAAAGAAAGGCCGCCAGACGACTCATATCGTATTTTCCTTCCGCGATATCACTTCCATGACGACAGGATAGTCTGAGGGTTATCTGTCACAGATTTGAGGGTGGTTCGTCACATTTGTTCTGACCTACTGAGGGTAATTTGTCACAGTTTTGCTGTTTCCTTCAGCCTGCATGGATTTTCTCATACTTTTTGAACTGTAATTTTTAAGGAAGCCAAATTTGAGGGCAGTTTGTCACAGTTGATTTCCTTCTCTTTCCCTTCGTCATGTGACCTGATATCGGGGGTTAGTTCGTCATCATTGATGAGGGTTGATTATCACAGTTTATTACTCTGAATTGGCTATCCGCGTGTGTACCTCTACCTGGAGTTTTTCCCACGGTGGATATTTCTTCTTGCGCTGAGCGTAAGAGCTATCTGACAGAACAGTTCTTCTTTGCTTCCTCGCCAGTTCGCTCGCTATGCTCGGTTACACGGCTGCGGCGAGCGCTAGTGATAATAAGTGACTGAGGTATGTGCTCTTCTTATCTCCTTTTGTAGTGTTGCTCTTATTTTAAACAACTTTGCGGTTTTTTGATGACTTTGCGATTTTGTTGTTGCTTTGCAGTAAATTGCAAGATTTAATAAAAAAACGCAAAGCAATGATTAAAGGATGTTCAGAATGAAACTCATGGAAACACTTAACCAGTGCATAAACGCTGGTCATGAAATGACGAAGGCTATCGCCATTGCACAGTTTAATGATGACAGCCCGGAAGCGAGGAAAATAACCCGGCGCTGGAGAATAGGTGAAGCAGCGGATTTAGTTGGGGTTTCTTCTCAGGCTATCAGAGATGCCGAGAAAGCAGGGCGACTACCGCACCCGGATATGGAAATTCGAGGACGGGTTGAGCAACGTGTTGGTTATACAATTGAACAAATTAATCATATGCGTGATGTGTTTGGTACGCGATTGCGACGTGCTGAGGACGTATTTCCACCGGTGATCGGGGTTGCTGCCCATAAAGGTGGCGTTTACAAAACCTCAGTTTCTGTTCATCTTGCTCAGGATCTGGCTCTGAAGGGGCTACGTGTTTTGCTCGTGGAAGGTAACGACCCCCAGGGAACAGCCTCAATGTATCACGGATGGGTACCAGATCTTCATATTCATGCAGAAGATACTCTCCTGCCTTTCTATCTTGGGGAAAAGGACGATGTCACTTATGCAATAAAGCCCACTTGCTGGCCGGGGCTTGACATTATTCCTTCCTGTCTGGCTCTGCACCGTATTGAAACTGAGTTAATGGGCAAATTTGATGAAGGTAAACTGCCCACCGATCCACACCTGATGCTCCGACTGGCCATTGAAACTGTTGCTCATGACTATGATGTCATAGTTATTGACAGCGCGCCTAACCTGGGTATCGGCACGATTAATGTCGTATGTGCTGCTGATGTGCTGATTGTTCCCACGCCTGCTGAGTTGTTTGACTACACCTCCGCACTGCAGTTTTTCGATATGCTTCGTGATCTGCTCAAGAACGTTGATCTTAAAGGGTTCGAGCCTGATGTACGTATTTTGCTTACCAAATACAGCAATAGTAATGGCTCTCAGTCCCCGTGGATGGAGGAGCAAATTCGGGATGCCTGGGGAAGCATGGTTCTAAAAAATGTTGTACGTGAAACGGATGAAGTTGGTAAAGGTCAGATCCGGATGAGAACTGTTTTTGAACAGGCCATTGATCAACGCTCTTCAACTGGTGCCTGGAGAAATGCTCTTTCTATTTGGGAACCTGTCTGCAATGAAATTTTCGATCGTCTGATTAAACCACGCTGGGAGATTAGATAATGAAGCGTGCGCCTGTTATTCCAAAACATACGCTCAATACTCAACCGGTTGAAGATACTTCGTTATCGACACCAGCTGCCCCGATGGTGGATTCGTTAATTGCGCGCGTAGGAGTAATGGCTCGCGGTAATGCCATTACTTTGCCTGTATGTGGTCGGGATGTGAAGTTTACTCTTGAAGTGCTCCGGGGTGATAGTGTTGAGAAAACCTCTCGGGTATGGTCAGGTAATGAACGTGACCAGGAGCTGCTTACTGAGGACGCACTGGATGATCTCATCCCTTCTTTTCTACTGACTGGTCAACAGACACCGGCGTTCGGTCGAAGAGTATCTGGTGTCATAGAAATTGCCGATGGGAGTCGCCGTCGTAAAGCTGCTGCACTTACCGAAAGTGATTATCGTGTTCTGGTTGGCGAGCTGGATGATGAGCAGATGGCTGCATTATCCAGATTGGGTAACGATTATCGCCCAACAAGTGCTTATGAACGTGGTCAGCGTTATGCAAGCCGATTGCAGAATGAATTTGCTGGAAATATTTCTGCGCTGGCTGATGCGGAAAATATTTCACGTAAGATTATTACCCGCTGTATCAACACCGCCAAATTGCCTAAATCAGTTGTTGCTCTTTTTTCTCACCCCGGTGAACTATCTGCCCGGTCAGGTGATGCACTTCAAAAAGCCTTTACAGATAAAGAGGAATTACTTAAGCAGCAGGCATCTAACCTTCATGAGCAGAAAAAAGCTGGGGTGATATTTGAAGCTGAAGAAGTTATCACTCTTTTAACTTCTGTGCTTAAAACGTCATCTGCATCAAGAACTAGTTTAAGCTCACGACATCAGTTTGCTCCTGGAGCGACAGTATTGTATAAGGGCGATAAAATGGTGCTTAACCTGGACAGGAGCCGTGTTCCAACTGAGTGTATAGAGAAAATTGAGGCCATTCTTAAGGAACTTGAAAAGCCAGCACCCTGATGCGACCACGTTTTAGTCTACGTTTATCTGTCTTTACTTAATGTCCTTTGTTACAGGCCAGAAAGCATAACTGGCCTGAATATTCTCTCTGGGCCCACTGTTCCACTTGTATCGTCGGTCTGATAATCAGACTGGGACCACGGTCCCACTCGTATCGTCGGTCTGATTATTAGTCTGGGACCACGGTCCCACTCGTATCGTCGGTCTGATTATTAGTCTGGGACCACGGTCCCACTCGTATCGTCGGTCTGATAATCAGACTGGGACCACGGTCCCACTCGTATCGTCGGTCTGATTATTAGTCTGGGACCATGGTCCCACTCGTATCGTCGGTCTGATTATTAGTCTGGGACCACGGTCCCACTCGTATCGTCGGTCTGATTATTAGTCTGGAACCACGGTCCCACTCGTATCGTCGGTCTGATTATTAGTCTGGGACCACGGTCCCACTCGTATCGTCGGTCTGATTATTAGTCTGGGACCACGATCCCACTCGTGTTGTCGGTCTGATTATCGGTCTGGGACCACGGTCCCACTTGTATTGTCGATCAGACTATCAGCGTGAGACTACGATTCCATCAATGCCTGTCAAGGGCAAGTATTGACATGTCGTCGTAACCTGTAGAACGGAGTAACCTCGGTGTGCGGTTGTATGCCTGCTGTGGATTGCTGCTGTGTCCTGCTTATCCACAACATTTTGCGCACGGTTATGTGGACAAAATACCTGGTTACC +R6K: + description: Medium copy (15-30 copies) or high copy depending on Pi expression. Requires Pi protein. Orthogonal to ColE2. + sequence: GATCTGAAGATCAGCAGTTCAACCTGTTGATAGTACGTACTAAGCTCTCATGTTTCACGTACTAAGCTCTCATGTTTAACGTACTAAGCTCTCATGTTTAACGAACTAAACCCTCATGGCTAACGTACTAAGCTCTCATGGCTAACGTACTAAGCTCTCATGTTTCACGTACTAAGCTCTCATGTTTGAACAATAAAATTAATATAAATCAGCAACTTAAATAGCCTCTAAGGTTTTAAGTTTTATAAGAAAAAAAAGAATATATAAGGCTTTTAAAGCCTTTAAGGTTTAACGGTTGTGGACAACAAGCCAGGGATGTAACGCACTGAGAAGCCCTTAGAGCCTCTCAAAGCAATTTTGAGTGACACAGGAACACTTAACGGCTGACATGG +CloDF13: + description: Medium copy (20-40 copies) Escherichia coli origin. Compatible with pUC/pBR322, p15A, RSF1030, ColA, and SC101 + sequence: GATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTTTTGCCCTGTAAACGAAAAAACCACCTGGGGAGGTGGTTTGATCGAAGGTTAAGTCAGTTGGGGAACTGCTTAACCGTGGTAACTGGCTTTCGCAGAGCACAGCAACCAAATCTGTCCTTCCAGTGTAGCCGGACTTTGGCGCACACTTCAAGAGCAACCGCGTGTTTAGCTAAACAAATCCTCTGCGAACTCCCAGTTACCAATGGCTGCTGCCAGTGGCGTTTTACCGTGCTTTTCCGGGTTGGACTCAAGTGAACAGTTACCGGATAAGGCGCAGCAGTCGGGCTGAACGGGGAGTTCTTGCTTACAGCCCAGCTTGGAGCGAACGACCTACACCGAGCCGAGATACCAGTGTGTGAGCTATGAGAAAGCGCCACACTTCCCGTAAGGGAGAAAGGCGGAACAGGTATCCGGTAAACGGCAGGGTCGGAACAGGAGAGCGCAAGAGGGAGCGACCCGCCGGAAACGGTGGGGATCTTTAAGTCCTGTCGGGTTTCGCCCGTACTGTCAGATTCATGGTTGAGCCTCACGGCTCCCACAGATGCACCGGAAAAGCGTCTGTTTATGTGAACTCTGGCAGGAGGGCGGAGCCTATGGAAAAACGCCACCGGCGCGGCCCTGCTGTTTTGCCTCACATGTTAGTCCCCTGCTTATCCACGGAATCTGTGGGTAACTTTGTATGTGTCCGCAGCGC +pLG13: + description: pLG13 is a ColE1 family plasmid that natively contains the EcoRV restriction-modification system + sequence: TTGAGATCCTTTTTTTTGCGCGCAATCTGCTGTCTGTAGACGAAAAAACCACCCTGGTAGGTGGTTTTTCGAAGGTTAAATAATCCTGGCAGATTATTTAACCGTGGTAACAGGGTGTACAAGACCGCTGCCACCAAATCTGTCCTTTCAGTGTAGCCGCAGTTGGTCCTTCACTTCAAGAACTACGTATCAGCAATTCTTGTACATCCTCTACCAGTGGCTGCTGCCAGTGGCGTTAAGGCGTGACTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCAACCCGAAGGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGGTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA +pSCANS-inducible: + description: Inducible copy number vector. Sourced from pOpen_v4 + sequence: agagcttggcaggtgtcgcgtatgcatcgatcacgtgtccacgttctttaatagtggactcttgttccaaactggaacaacactcggatcgatccggcgcgcaccgtgggaaaaactccaggtagaggtacacacgcggatagccaattcagagtaataaactgtgataatcaaccctcatcaatgatgacgaactaacccccgatatcaggtcacatgacgaagggaaagagaaggaaatcaactgtgacaaactgccctcaaatttggcttccttaaaaattacagttcaaaaagtatgagaaaatccatgcaggctgaaggaaacagcaaaactgtgacaaattaccctcagtaggtcagaacaaatgtgacgaaccaccctcaaatctgtgacagataaccctcagactatcctgtcgtcatggaagtgatatcgcggaaggaaaatacgatatgagtcgtctggcggcctttctttttctcaatgtatgagaggcgcattggagttctgctgttgatctcattaacacagacttgcaggaagcggcggcggaagtcaggcatacgctggtaactttgaggcagctggtaacgctctatgatccagtcgattttcagagacacgatgcctgagccatccggcttacgatactgacacagggattcgtataaacgcatggcatacggattggtgatttcttttgtttcactaagccgaaactgcgtaaaccggttctgtaacccgataaagaagggaatgagatatgggttgatatgtacactgtaaagccctctggatggactgtgcgcacgtttgataaaccaaggaaaagattcatagcctttttcatcaccggcatcctcttcagggcgataaaaaaccacttccttccccgcgaaactcttcaatgcctgccgtatatccttactggcttccgcagaggtcaatccgaatatttcagcatatttagcaacatggatctcgcagataccgtcatgttcctgtagggtgccatcagattttctgatctggtcaacgaacagatacagcatacgtttttgatcccgggagagactatatgccgcctcagtgaggtcgtttgactggacgattcgcgggctatttttacgtttcttgtgattgataaccgctgtttccgccatgacagatccatgtgaagtgtgacaagtttttagattgtcacactaaataaaaaagagtcaataagcagggataactttgtgaaaaaacagcttcttctgagggcaatttgtcacagggttaagggcaatttgtcacagacaggactgtcatttgagggtgatttgtcacactgaaagggcaatttgtcacaacaccttctgtagaaccagcatggataaaggcctacaaggcgctctaaaaaagaagatctaaaaactataaaaaaaataattataaaaatatccccgtggataagtggataaccccaagggaagttttttcaggcatcgtgtgtaagcagaatatataagtgctgttccctggtgcttcctcgctcactcgaaattcccggggatagctttatgcttgtaaaccgttttgtgaaaaaatttttaaaataaaaaaggggacctctagggtccccaattaattagtaatataatctattaaaggtcattcaaaaggtcatccaccggatcaattcccctgctcgcgcaggctgggtgccaagctctcgggtaacatcaaggcccgatccttggagcccttgccctcccgcacgatgatcgtgccgtgatcgaaatccagatccttgacccgcagttgcaaaccctcactgatccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatgtgagttagctcactcattaggcaccccaggctttacactttatgcttccggctcgtatgttgtgtggaattgtgagcggataacaatttcacacaggaaacagctatgaccatgattacgccaagcgcggtacctgtcgcggcaacgcgctaacagacgtagtaagaaccaccagcattgtaatgctggctaaagtcactttcctgagctgtataacgatgagcgattttactttttctggctatgaattggcctgctttgtaacacactccggtctatcccgtagcgccgggcatatcctgtcgcaatgtgcaaatctcgcggcaacaaccagtgaatacttcattcacaagcctcaccgcctgatcgcggcagaaactggttatagccaatcaaccgtcgttcgtgcattccgtgaagctgtaaacaaaggtatcctgtctgtagagattgttatcggcgatcaccgtgaacgtcgcgctaacctgtaccggtttacaccatcctttttggccttcgcacaacaagccaaaaatgcgctgatagaaagcaaattaaagatctcttcagcggcaaccaaggttaaagctgttctcgctaagacattggctttatttaattttttatccacacccccatgtcaaaatgataccccctccccctgtcaggatgacgtggcaataaagaataagaagtcacaagttaaaaaaacaaaaagatcagtttccggcggtgccggaacaaccagcctcaaaaaattgacttcatggatcgctaaggcaaaagcaaaggctgacaatctgcggttatccaaaaaacgcactcaaaaacatgagttcaagcagaaagtagaggcggctgcgcggaaatatgcttacctgaagaacaagcgttcgcctgatattggcgggatatcaaacttcgataacctaccgcattgcatgacggtaaacgaagctcttaatgcggttttagccaaaaataaagataacgaacaatggggtataccggcaggattcagagggtaatgaattgctctaattataaccatgcatactttcaacacctctagtttgccatgaggcaaactcataggtgtcctggtaagaggacactgttgccaaaactggacgcc diff --git a/parts/parts/ecoli/promoters.yaml b/parts/parts/ecoli/promoters.yaml new file mode 100644 index 0000000..e6f5a49 --- /dev/null +++ b/parts/parts/ecoli/promoters.yaml @@ -0,0 +1,101 @@ +# Anderson +BBa_J23119: + prefix: TACA + suffix: AACT + tags: ['promoter', 'anderson_promoter'] + sequence: ttgacagctagctcagtcctaggtataatgctagc +BBa_J23100: + prefix: TACA + suffix: AACT + tags: ['promoter', 'anderson_promoter'] + sequence: ttgacggctagctcagtcctaggtacagtgctagc +BBa_J23101: + prefix: TACA + suffix: AACT + tags: ['promoter', 'anderson_promoter'] + sequence: tttacagctagctcagtcctaggtattatgctagc +BBa_J23102: + prefix: TACA + suffix: AACT + tags: ['promoter', 'anderson_promoter'] + sequence: ttgacagctagctcagtcctaggtactgtgctagc +BBa_J23103: + prefix: TACA + suffix: AACT + tags: ['promoter', 'anderson_promoter'] + sequence: ctgatagctagctcagtcctagggattatgctagc +BBa_J23104: + prefix: TACA + suffix: AACT + tags: ['promoter', 'anderson_promoter'] + sequence: ttgacagctagctcagtcctaggtattgtgctagc +BBa_J23105: + prefix: TACA + suffix: AACT + tags: ['promoter', 'anderson_promoter'] + sequence: tttacggctagctcagtcctaggtactatgctagc +BBa_J23106: + prefix: TACA + suffix: AACT + tags: ['promoter', 'anderson_promoter'] + sequence: tttacggctagctcagtcctaggtatagtgctagc +BBa_J23107: + prefix: TACA + suffix: AACT + tags: ['promoter', 'anderson_promoter'] + sequence: tttacggctagctcagccctaggtattatgctagc +BBa_J23108: + prefix: TACA + suffix: AACT + tags: ['promoter', 'anderson_promoter'] + sequence: ctgacagctagctcagtcctaggtataatgctagc +BBa_J23109: + prefix: TACA + suffix: AACT + tags: ['promoter', 'anderson_promoter'] + sequence: tttacagctagctcagtcctagggactgtgctagc +BBa_J23110: + prefix: TACA + suffix: AACT + tags: ['promoter', 'anderson_promoter'] + sequence: tttacggctagctcagtcctaggtacaatgctagc +BBa_J23111: + prefix: TACA + suffix: AACT + tags: ['promoter', 'anderson_promoter'] + sequence: ttgacggctagctcagtcctaggtatagtgctagc +BBa_J23112: + prefix: TACA + suffix: AACT + tags: ['promoter', 'anderson_promoter'] + sequence: ctgatagctagctcagtcctagggattatgctagc +BBa_J23113: + prefix: TACA + suffix: AACT + tags: ['promoter', 'anderson_promoter'] + sequence: ctgatggctagctcagtcctagggattatgctagc +BBa_J23114: + prefix: TACA + suffix: AACT + tags: ['promoter', 'anderson_promoter'] + sequence: tttatggctagctcagtcctaggtacaatgctagc +BBa_J23115: + prefix: TACA + suffix: AACT + tags: ['promoter', 'anderson_promoter'] + sequence: tttatagctagctcagcccttggtacaatgctagc +BBa_J23116: + prefix: TACA + suffix: AACT + tags: ['promoter', 'anderson_promoter'] + sequence: ttgacagctagctcagtcctagggactatgctagc +BBa_J23117: + prefix: TACA + suffix: AACT + tags: ['promoter', 'anderson_promoter'] + sequence: ttgacagctagctcagtcctagggattgtgctagc +BBa_J23118: + prefix: TACA + suffix: AACT + tags: ['promoter', 'anderson_promoter'] + sequence: ttgacggctagctcagtcctaggtattgtgctagc diff --git a/parts/parts/pichia_toolkit.yaml b/parts/parts/pichia_toolkit.yaml new file mode 100644 index 0000000..032a00d --- /dev/null +++ b/parts/parts/pichia_toolkit.yaml @@ -0,0 +1,114 @@ +PARS: + description: placeholder + tags: [origin] + prefix: GTCT + suffix: AAGC + sequence: cgataagctgggggaacattcgcgaaaatgaaacaagtcggctgttatagtatatttattataatattgaaagatctcaaaagactacttatttttgaatgaaccaagtatgaaatcaacctatttggggttgaccaaaataagtaaatattaattggatccgcacaaacgaagg +Pichia_Bxb1: + description: placeholder + tags: [origin] + prefix: GTCT + suffix: AAGC + sequence: tggccgtggccgtgctcgtcctcgtcggccggcttgtcgacgacggcggtcaccgtcgtcaggatcatccgggccacaagcttgctgacagaagcctcaagaaaaaaaaaattcttcttcgactatgctggaggcagagatgatcgagccggtagttaactatatatagctaaattggttccatcac +tAOX1_terminator: + description: placeholder + tags: [terminator] + prefix: ATCCTAA + suffix: CGCT + sequence: tcaagaggatgtcagaatgccatttgcctgagagatgcaggcttcatttttgatacttttttatttgtaacctatatagtataggattttttttgtcattttgtttcttctcgtacgagcttgctcctgatcagcctatctcgcagctgatgaatatcttgtggtaggggtttgggaaaatcattcgagtttgatgtttttcttggtatttcccactcctcttcagagtacagaagattaagtgaga +tAOX1_terminator_ctag: + description: placeholder + tags: [terminator_ctag] + prefix: ATGT + suffix: CGCT + sequence: tcaagaggatgtcagaatgccatttgcctgagagatgcaggcttcatttttgatacttttttatttgtaacctatatagtataggattttttttgtcattttgtttcttctcgtacgagcttgctcctgatcagcctatctcgcagctgatgaatatcttgtggtaggggtttgggaaaatcattcgagtttgatgtttttcttggtatttcccactcctcttcagagtacagaagattaagtgaga +RFP(Ppastoris): + description: placeholder + tags: [cds] + prefix: A + suffix: GGATCC + sequence: atggcaacttccggtatggtgtcaaagggagaggaaaataatatggctattattaaggagtttatgcgttttaaggtacatatggaaggttctgtcaacggtcacgaattcgaaattgaaggtgagggggaggggaggccatacgagggaactcagactgctaagttaaaggtcactaaaggtggtcctttacctttcgcctgggatatcctgtctccacagtttatgtacggttcaaaggcttatgtgaaacatcctgccgatatcccagattatcttaaactttctttccctgagggttttaagtgggagagggtaatgaactttgaagacggtggtgtggtcactgttactcaggactcaagtctgcaggacggtgagttcatctacaaggtgaagctgagaggtaccaattttccatcagatggtcccgtgatgcaaaaaaagacaatgggttgggaagcttctagtgaacgtatgtatcccgaagatggagctttgaaaggtgaaattaagcaaagactaaaacttaaggatggtggacattacgatgctgaagttaagacgacctacaaggccaaaaagccagtccagttgcctggagcatacaatgttaacatcaaattggatataacttcccataatgaagactataccatcgtcgagcaatacgaacgagccgaagggagacacagtactggtggtatggatgaactttataaaggatccggaaccgca +EGFP(Ppastoris): + description: placeholder + tags: [cds] + prefix: A + suffix: GGATCC + sequence: atggtgagcaagggcgaggagctgttcaccggggtggtgcccatcctggtcgagctggacggcgacgtaaacggccacaagttcagcgtgtccggcgagggcgagggcgatgccacctacggcaagctgaccctgaagttcatctgcaccaccggcaagctgcccgtgccctggcccaccctcgtgaccaccctgacctacggcgtgcagtgcttcagccgctaccccgaccacatgaagcagcacgacttcttcaagtccgccatgcccgaaggctacgtccaggagcgcaccatcttcttcaaggacgacggcaactacaagacccgcgccgaggtgaagttcgagggcgacaccctggtgaaccgcatcgagctgaagggcatcgacttcaaggaggacggcaacatcctggggcacaagctggagtacaactacaacagccacaacgtctatatcatggccgacaagcagaagaacggcatcaaggtgaacttcaagatccgccacaacatcgaggacggcagcgtgcagctcgccgaccactaccagcagaacacccccatcggcgacggccccgtgctgctgcccgacaaccactacctgagcacccagagcgccctgagcaaagaccccaacgagaagcgcgatcacatggtcctgctggagttcgtgaccgccgccgggatcactctcggcatggacgagctgtacaag +ntag_pichia_alphaAmylase-alphaMFDelta: + description: placeholder + tags: [ntag] + prefix: AACT + suffix: AATG + sequence: atggtggcatggtggtccttattcttatatggtcttcaagttgctgctcctgcccttgctatgagatttcctagtattttcactgctgtgctatttgccgctagttccgctctagctgctccagttaatactactactgaagatgaattggagggtgacttcgatgttgctgttctgcctttttccgcttctatcgcagccaaggaagaaggtgtatctctagagaagcgtgg +ntag_pichia_killer-alphaMFDelta: + description: placeholder + tags: [ntag] + prefix: AACT + suffix: AATG + sequence: atgaccaaaccaacgcaagtcttagttcgttcagtctctattttattcttcatcacactgttgcacttggttgttgcaatgagatttcctagtattttcactgctgtgctatttgccgctagttccgctctagctgctccagttaatactactactgaagatgaattggagggtgacttcgatgttgctgttctgcctttttccgcttctatcgcagccaaggaagaaggtgtatctctagagaagcgtgg +ntag_pichia_invertase-alphaMFDelta: + escription: placeholder + tags: [ntag] + prefix: AACT + suffix: AATG + sequence: atgttattgcaagcttttttatttctgctggcaggttttgcagcaaagatttctgccatgagatttcctagtattttcactgctgtgctatttgccgctagttccgctctagctgctccagttaatactactactgaagatgaattggagggtgacttcgatgttgctgttctgcctttttccgcttctatcgcagccaaggaagaaggtgtatctctagagaagcgtgg +ntag_pichia_inulinase-alphaMFDelta: + description: placeholder + tags: [ntag] + prefix: AACT + suffix: AATG + sequence: atgaaactggcttactccctgttgctacctctggctggagtttccgctatgagatttcctagtattttcactgctgtgctatttgccgctagttccgctctagctgctccagttaatactactactgaagatgaattggagggtgacttcgatgttgctgttctgcctttttccgcttctatcgcagccaaggaagaaggtgtatctctagagaagcgtgg +ntag_pichia_serumalbumin-alphaMFDelta: + description: placeholder + tags: [ntag] + prefix: AACT + suffix: AATG + sequence: atgaagtgggtaactttcatctcattgttattcttgttctcctctgcttactctatgagatttcctagtattttcactgctgtgctatttgcctctagttccgctctagctgctccagttaatactactactgaagatgaattggagggtgacttcgatgttgctgttctgcctttttccgcttctatcgcagccaaggaagaaggtgtatctctagagaagcgtgg +ntag_pichia_glucoamylase-alphaMFDelta: + description: placeholder + tags: [ntag] + prefix: AACT + suffix: AATG + sequence: atgtctttcagatccctattggcattgtcagggttggtctgttctggattggctatgagatttcctagtattttcactgctgtgctatttgccgctagttccgctctagctgctccagttaatactactactgaagatgaattggagggtgacttcgatgttgctgttctgcctttttccgcttctatcgcagccaaggaagaaggtgtatctctagagaagcgtgg +ntag_pichia_alphaMFDelta-noKex: + description: placeholder + tags: [ntag] + prefix: AACT + suffix: AATG + sequence: atgagattcccatcaatttttactgctgttctgttcgccgcttctagtgcacttgccatgagatttcctagtattttcactgctgtgctatttgccgctagttccgctctagctgctccagttaatactactactgaagatgaattggagggtgacttcgatgttgctgttctgcctttttccgcttctatcacagccaaggaagaaggtgtatctctagagaagcgtgg +ntag_pichia_alphaMFDelta: + description: placeholder + tags: [ntag] + prefix: AACT + suffix: AATG + sequence: atgagatttcctagtattttcactgctgtgctatttgccgctagttccgctctagctgctccagttaatactactactgaagatgaattggagggtgacttcgatgttgctgttctgcctttttccgcttctatcgcagccaaggaagaaggtgtatctctagagaagcgtgg +ntag_pichia_alphaMF-noEAEA: + description: placeholder + tags: [ntag] + prefix: AACT + suffix: AATG + sequence: atgagatttccttcaatttttactgctgttttattcgcagcatcctccgcattagctgctccagtcaacactacaacagaagatgaaacggcacaaattccggctgaagctgtcatcggttactcagatttagaaggggatttcgatgttgctgttttgccattttccaacagcacaaataacgggttattgtttataaatactactattgccagcattgctgctaaagaagaaggggtatctctcgagaaaagagg +ntag_pichia_alphaMF: + description: placeholder + tags: [ntag] + prefix: AACT + suffix: AATG + sequence: atgagatttccttcaatttttactgctgttttattcgcagcatcctccgcattagctgctccagtcaacactacaacagaagatgaaacggcacaaattccggctgaagctgtcatcggttactcagatttagaaggggatttcgatgttgctgttttgccattttccaacagcacaaataacgggttattgtttataaatactactattgccagcattgctgctaaagaagaaggggtatctctcgagaaaagagaggctgaagctgg +pTPI1_promoter: + description: "Triose phosphate isomerase 1 promoter" + tags: [promoter] + prefix: TACA + suffix: AACT + sequence: gtgtttaaagattacggatatttaacttacttagaataatgccatttttttgagttataataatcctacgttagtgtgagcgggatttaaactgtgaggaccttaatacattcagacacttctgcggtatcaccctacttattcccttcgagattatatctaggaacccatcaggttggtggaagattacccgttctaagacttttcagcttcctctattgatgttacacctggacaccccttttctggcatccagtttttaatcttcagtggcatgtgagattctccgaaattaactaaagcaatcacacaattctctcggataccacctcggttgaaactgacaggtggtttgttacgcatgctaatgcaaaggagcctatatacctttggctcggctgctgtaacagggaatataaagggcagcataatttaggagtttagtgaacttgcaacatttactattttcccttcttacgtaaatatttttctttttaattctaaatcaatctttttcaattttttgtttgtattcttttcttgcttaaatctataactacaaaaaacacatacataaactaaaa +pENO1_promoter: + description: "Enolase 1 promoter" + tags: [promoter] + prefix: TACA + suffix: AACT + sequence: agaaagcatactatactattcgacattcctttcaatcctggaattaacagtcacttttaaaaaagacatctaccgtgaaggtgccgtagagtatcgcgttaccatatcgccaaaaactgatatacgccgcggaaaccaggcaaacaattgaaaagaaaaattttgaggaactctctgcatcgaagccgtctagagttaccactagtcagatgccgcgggcacttgagcacctcatgcacagcaataacacaacacaatggttagtagcaacctgaattcggtcattgatgcatgcatgtgccgtgaagcgggacaaccagaaaagtcgtctataaatgccggcacgtgcgatcatcgtggcggggttttaagagtgcatatcacaaattgtcgcattaccgcggaaccgccagatattcattacttgacgcaaaagcgtttgaaataatgacgaaaaagaaggaagaaaaaaaaagaaaaataccgcttctaggcgggttatctactgatccgagcttccactaggatagcacccaaacacctgcatatttggacgacctttacttacaccaccaaaaaccactttcgcctctcccgcccctgataacgtccactaattgagcgattacctgagcggtcctcttttgtttgcagcatgagacttgcatactgcaaatcgtaagtagcaacctctcaaggtcaaaactgtatggaaaccttgtcacctcacttaattctagctagcctaccctgcaagtcaagagctctccgtgattcctagccacctcaaggtatgcctctccccggaaactgtggccttttctggcacacatgatctccacgatttcaacatataaatagcttttgataatggcaatattaatcaaatttattttacttctttcttgtaacatctctcttgtaatcccttattccttctagctatttttcataaaaaaccaagcaactgcttatcaacacacaaacactaaatcaaa +pAOX1_promoter: + description: "Alcohol oxidase 1 promoter" + tags: [promoter] + prefix: TACA + suffix: AACT + sequence: gatctaacatccaaagacgaaaggttgaatgaaacctttttgccatccgacatccacaggtccattctcacacataagtgccaaacgcaacaggaggggatacactagcagcagaccgttgcaaacgcaggacctccactcctcttctcctcaacacccacttttgccatcgaaaaaccagcccagttattgggcttgattggagctcgctcattccaattccttctattaggctactaacaccatgactttattagcctgtctatcctggcccccctggcgaggttcatgtttgtttatttccgaatgcaacaagctccgcattacacccgaacatcactccagatgagggctttctgagtgtggggtcaaatagtttcatgttccccaaatggcccaaaactgacagtttaaacgctgtcttggaacctaatatgacaaaagcgtgatctcatccaagatgaactaagtttggttcgttgaaatgctaacggccagttggtcaaaaagaaacttccaaaagtcggcataccgtttgtcttgtttggtattgattgacgaatgctcaaaaataatctcattaatgcttagcgcagtctctctatcgcttctgaaccccggtgcacctgtgccgaaacgcaaatggggaaacacccgctttttggatgattatgcattgtctccacattgtatgcttccaagattctggtgggaatactgctgatagcctaacgttcatgatcaaaatttaactgttctaacccctacttgacagcaatatataaacagaaggaagctgccctgtcttaaacctttttttttatcatcattattagcttactttcataattgcgactggttccaattgacaagcttttgattttaacgacttttaacgacaacttgagaagatcaaaaaacaactaattattcgaaacg diff --git a/parts/parts/vnat.yaml b/parts/parts/vnat.yaml new file mode 100644 index 0000000..7664aba --- /dev/null +++ b/parts/parts/vnat.yaml @@ -0,0 +1,178 @@ +#https://doi.org/10.1021/acssynbio.1c00126 +dummy_promoter: + prefix: TACA + suffix: AACT + sequence: N +Ptrc_promoter: + prefix: TACA + suffix: AACT + sequence: N +Ptet_promoter: + prefix: TACA + suffix: AACT + sequence: N +Para(Vn)_promoter: + prefix: TACA + suffix: AACT + sequence: N +Prham(Vn)_promoter: + prefix: TACA + suffix: AACT + sequence: N +# dropout sfGFP / mScarlet for each +B0029_rbs: + prefix: AACT + suffix: AATG + sequence: N +B0030_rbs: + prefix: AACT + suffix: AATG + sequence: N +B0031_rbs: + prefix: AACT + suffix: AATG + sequence: N +B0032_rbs: + prefix: AACT + suffix: AATG + sequence: N +B0033_rbs: + prefix: AACT + suffix: AATG + sequence: N +B0034_rbs: + prefix: AACT + suffix: AATG + sequence: N +B0035_rbs: + prefix: AACT + suffix: AATG + sequence: N +B0064_rbs: + prefix: AACT + suffix: AATG + sequence: N +# cds +lux_operon(Vn): + prefix: A + suffix: ATCC + sequence: N +azurite(Vn): + prefix: A + suffix: ATCC + sequence: N +mTurquoise(Vn): + prefix: A + suffix: ATCC + sequence: N +sfGFP(Vn): + prefix: A + suffix: ATCC + sequence: N +mVenus(Vn): + prefix: A + suffix: ATCC + sequence: N +mScarlet-I(Vn): + prefix: A + suffix: ATCC + sequence: N +mCherry(Vn): + prefix: A + suffix: ATCC + sequence: N +mKate-2(Vn): + prefix: A + suffix: ATCC + sequence: N +# terminators -- make ctag versions as well +B0010_terminator: + prefix: ATCCTAA + suffix: CGCT + sequence: N +B0015_terminator: + prefix: ATCCTAA + suffix: CGCT + sequence: N +B1002_terminator: + prefix: ATCCTAA + suffix: CGCT + sequence: N +B1003_terminator: + prefix: ATCCTAA + suffix: CGCT + sequence: N +B1004_terminator: + prefix: ATCCTAA + suffix: CGCT + sequence: N +B1006_terminator: + prefix: ATCCTAA + suffix: CGCT + sequence: N +B1007_terminator: + prefix: ATCCTAA + suffix: CGCT + sequence: N +B1009_terminator: + prefix: ATCCTAA + suffix: CGCT + sequence: N +B1010_terminator: + prefix: ATCCTAA + suffix: CGCT + sequence: N +# n and c tag +# ntags: 3xFlag, 6xHis, Azurite, mTurqoise, sfGFP, mVenus, mScarlet-I, mCherry, mKate-2 +3xFlag(Vn)_ctag: + prefix: ATCC + suffix: ATGT + sequence: N +6xHis(Vn)_ctag: + prefix: ATCC + suffix: ATGT + sequence: N +Azurite(Vn)_ctag: + prefix: ATCC + suffix: ATGT + sequence: N +mTurquoise(Vn)_ctag: + prefix: ATCC + suffix: ATGT + sequence: N +sfGFP(Vn)_ctag: + prefix: ATCC + suffix: ATGT + sequence: N +mVenus(Vn)_ctag: + prefix: ATCC + suffix: ATGT + sequence: N +mScarlet-I(Vn)_ctag: + prefix: ATCC + suffix: ATGT + sequence: N +mCherry_ctag: + prefix: ATCC + suffix: ATGT + sequence: N +mKate-2_ctag: + prefix: ATCC + suffix: ATGT + sequence: N +I11012_ctag: + prefix: ATCC + suffix: ATGT + sequence: N +M0050_ctag: + prefix: ATCC + suffix: ATGT + sequence: N +M0051_ctag: + prefix: ATCC + suffix: ATGT + sequence: N +M0052_ctag: + prefix: ATCC + suffix: ATGT + sequence: N diff --git a/parts/parts/yeast_toolkit.yaml b/parts/parts/yeast_toolkit.yaml new file mode 100644 index 0000000..dbcf2ff --- /dev/null +++ b/parts/parts/yeast_toolkit.yaml @@ -0,0 +1,252 @@ +CUP1_promoter: + description: placeholder + tags: [promoter] + prefix: TACA + suffix: AACT + sequence: atcttagccttgttgctagttagaaaaagacatttttgctgtcagtcactgtcaagagattcttttgctggcatttcttctaggagcaaaaagagcgatgcgtcttttccgctgaaccgttccagcaaaaaagactaccaacgcaatatggattgtcagaatcatataaaagagaagcaaataactccttgtcttgtatcaattgcattataatatcttcttgttagtgcaatatcatatagaagtcatcgaaatagatattaagaaaaacaaactgtacaatcaatcaatcaatcatcacataaaagatc +GAL1_promoter: + description: placeholder + tags: [promoter] + prefix: TACA + suffix: AACT + sequence: ccccattatcttagcctaaaaaaaccttctctttggaactttcagtaatacgcttaactgctcattgctatattgaagtacggattagaagccgccgagcgggcgacagccctccgacggatgactctcctccgtgcgtcctcgtcatcaccggtcgcgttcctgaaacgcagatgtgcctcgcgccgcactgctccgaacaataaagattctacaatactagcttttatggttatgaagaggaaaaattggcagtaacctggccccacaaaccttcaaattaacgaatcaaattaacaaccataggatgataatgcgattagttttttagccttatttctggggtaattaatcagcgaagcgatgatttttgatctattaacagatatataaatggaaaagctgcataaccactttaactaatactttcaacattttcagtttgtattacttcttattcaaatgtcataaaagtatcaacaaaaaattgttaatatacctctatactttaacgtcaaggagaaaaaactataagatc +MF(alpha)2_promoter: + description: placeholder + tags: [promoter] + prefix: TACA + suffix: AACT + sequence: ttgatctgccttttatagctaagaaaaaaaaagagaaaaaaataataagtgcgggtaacctcatgcattgtaaaacggtagggctacaaggcactacttactctacgctatagaagaagcttcaagacatggtgttgttcttatttgagcgaactcagctgtcaccgtttcgggaaactctatagttttctgcgtttcagtacgcagttgggcgtgctaaagttgttttcctaatttgctagtgttcacttgctcattgatgtccctaaaaggtaatgtcgcatagtaagatgtatgtcccatatcaaacgcattcctagaaaaacatgcactgaatgaataagcagttacttcgattactaatcttcacagcataatttttctcttatcttgtaaacctctttctatcttttcataaagtttgtcacttatatttcttcattggtacatcaatgccagcaacgatgtgcgcatctgggcgacgcctgtagtgattgttttcaaggtatcgagccaaactattcatcgttactgtttcaaatattcagttgtttcagtacagagtcgccgtggacctagtgaaacttggtgtctttacagcgcagagatgagggcttatatgtataaaagctgtccttgattctggtgtagtttgaggtgtccttcctatatctgtttttatattctatataatggataattactaccatcacctgcatcaaattccagtaaattcacatattggagaaaagatc +MFA1_promoter: + description: placeholder + tags: [promoter] + prefix: TACA + suffix: AACT + sequence: cgctagaaggaaattggaagaactaaatagagttctgaaaaacaatccatatttcaattcaaatgtgaacgtacacaattcgagactcaaagatgctgtaccgttcacgccgtttaacggtgatagagaagcacacccaaggtttacgttgaaaggttcagtatacaatgacccattcatcaaacatcttgagcacaggaaagaatttattgcgtctgggttcaacactaattatgcgtacgaaagggtgttgacagaggcatttatgggcttaggatgtgttatatccgaggagctttaaaacatcaggatagtgtgcaacgtggcataagctatgtaatcaactactttttattttctatgtacgcatatacatgcattcacgatctgtttcagtgttcagaaaaaaggcacctactgctacggttggcccatacctttattctttgttcttgttacaaacgagtgtgtaattacccaaaaaggaaatttacatgttaaatgaaacccagtaatcagaaaaaacagttaagaaacctaaaatggtagagataaagatacagattcagtggttgctgaaaatcaagtaaaaaaatgaaatagagtcatcatatataaaccgccagaaatgaattaatgagagggatctgtaactgtttctcggataaaaccaaaataagtacaaagccatcgaatagaaagatc +REV1_promoter: + description: placeholder + tags: [promoter] + prefix: TACA + suffix: AACT + sequence: gtgttgttatccgatacaaccggatatttttcttttaatgagtctaaaccgtgatagcttcaggttaatacaatcaaaaaaagctcaaatattcttttaatgccgcgttcacagattccaattgaatacaactaggtagttcattatatgaagcctttgctactatttttcactatagtctgccttcaccttaatgcagacatccacatattttaatcactttaaaataaaaaggaagatatattagaagctatgatccaatctgtaagccagattaaaattcacgaactcttctttcatttgaattgaatgctttgagttggggtagattatcgcaaattactcatcacatttattgactacgaacttgctgatgtcctttttttatttatatttttcttcagtgaagcgattttttttttacacagaccaagacggaaaaaagtagctaaggaagaaaacaaaatcatgaaaaaaatgtgaagtgatcatgcacatcgcatcaacttaaacattggcttagagatatatagagttagagtttacggcaacctttaagcaccaataccttttggcatagtctaaagacctggttcttaattttaaacaaatttaactaaagatttccctatcaaagaagtaacgagttgacagattttctcaaaataaatcgatactgcatttctaggcatatccagcgagatc +PSP2_promoter: + description: placeholder + tags: [promoter] + prefix: TACA + suffix: AACT + sequence: tgacccaacatcagatgacccaaggtccacctcttattaaaggacgtttgatccttcgacaccatggctctgttgaacttttatctgagagaggaaaaaaaggaaggaaaaaaaagaagaaacttcctttatttatttgtcttaaccacaacacacaatgcaataagatgcaatataatatcaaagccaatatcttatgttgctgatcctgagaaggaatatatacaatttatgtagtaaaataccttttcttctgcgagttgcaagaaatagaaaagactccgattgcgcatcgccagaataaaatttcacaaccacactttttggctgaactttttattacctgattaaacagagagagaaaaggtagaggtcaaaattttttaagcaaaactaaaaaagatgcaaaatcacgtgctgaaaatctaacataagggttaagattagagttttataggacttgttttgtaatatttcaaatacgagctaaccctactgatttcaattaggtctaatttagggttgagctgcactgaaatttcggaaattttgggttattttaaatgagacagaagaactacagagatacgttcttcagactttaaagcttatctccacaaagaattggtcaagaaatcatcctagaaaaacacgtttgctcactcgatcttaatcacatagagtgctggaacgggaagaaagatc +RAD27_promoter: + description: placeholder + tags: [promoter] + prefix: TACA + suffix: AACT + sequence: ccttgtgaaattgcaaatatggtgatttgaaacgtttcctagtgcagcaggatcacagataacgtgtaaagggcttagcagttgataatcctctctagttaagacctaaacaaaatgctgtcactaaccgtagtattaaatgacacactttggtgactttcgttaatggggatgtggtagtggccattgccaataaacaaaaagaacagggaaagaagtagaaagtgatataagtttgcttgccacttttcgtttttcacgaaaaaaacaggcgaaaaaaaatgctagacaagtacccggctgaatcacacctcgttaacagtgactttcggtgacagatacccgattgggcacccggctggtaagttatgatagaaagccaacgctgtactattggcttagctatggcaatattttgattatcagctagttttattaacgttataattagtgtaaccagtttttcatctatttcatttatttcatttatttactttaattgcagatccccctaacgcgtttaaagcttttattcactagcttatgtattttttataggaaacgcgacgcgtaacatcgcgcaaatgaaggttttgatgtattataatgaggtattcttccttatatacatcgatgaaaagcgttgacagcatacattggaaagaaataggaaacggacaccggaagaaaaaatagatc +POP6_promoter: + description: placeholder + tags: [promoter] + prefix: TACA + suffix: AACT + sequence: ttcgtgctttgtgataaagtgtttcacgtcatccgacatgacttcgtagttatggactgaactgtgtggtgaggttccatgatttcttaggtccagcagatacatgtctcttcccaatttcttgttaaggttacggccaatgcttcggttgttgagcttgttaccgaataagccgtgaagtatgataataggtggtcttggcttcccttcatccccagtttttactgcatctctcttgattatgtcatatgaaaggtccagtgggacttgcttttgttgcagcacctttgctaatgaatgaaaggcacatagtgactgcttaaaaatgcaggaacttaaattattccgaatggtattttgtctcacatatattgtcccatactgtgccaagatcccggctttacccagtatcatcattgtaccgttaccaattctcctcgtatatcacggttagtttttaaacctcggggtgacgtttactattggcgtactaatatattcttattttcttttcttttttgttggcagtttcaagcaacacatgtactggataaccaacccccgcacgctcttggaaaaaattgagaaggcatcggacacttgctgatgagtatttcgaaaaattccatgaagatgaggccaagattgtttggaagagattgaaaagaagaagaagaaaaaaagataaaagcaaatcaaaagatc +RNR2_promoter: + description: placeholder + tags: [promoter] + prefix: TACA + suffix: AACT + sequence: agtcgaacaagaagcaggcaaagtttagagcactgcccctccgcactcaaaaaagaaaaaactaggaggaaaataaaattctcaaccacacaaacacataaacacatacaaatacaaatacaagcttatttacttgacatcgcgcgatcttccactattcagcgccgtccgccctctctcgtgttttttgtttacgcgacaactatgcgaaatccggagcaacgggcaaccgtttggggaaagaccacacccacgcgcgatcgccatggcaacgaggtcgcacacgccccacacccagacctccctgcgagcgggcatgggtacaatgtccccgttgccacagacaccacttcgtagcacagcgcagagcgtagcgtgttgttgctgctgacaaaagaaaatttttcttagcaaagcaaaggaggggaagcacgggcagatagcaccgtaccatacccttggaaactcgaaatgaacgaagcaggaaatgagagaatgagagttttgtaggtatatatagcggtagtgtttgcgcgttaccatcatcttctggatctatctattgttcttttcctcatcactttcccctttttcgctcttcttcttgtcttttatttctttcttttttttaattgttccctcgattggctatctaccaaagaatccaaacttaatacacgtatttatttgtccaattaccagatc +SAC6_promoter: + description: placeholder + tags: [promoter] + prefix: TACA + suffix: AACT + sequence: tttgagaatgaccttccacgagctaaattgaaagggaagaatttattagttgaactcaagaaagaagaggatgacgtgggaaatggcatagaatcccttactaaatcgaacactaaactgaactccatgctggcgaacgaaggtaagatacacaaagctagtttccagaaaagtgtaaaatttaaactacctgataatatagtgactgaagaaaccgtggaacttaaagaaataaaggacttgctactacaaatgttgagatgacagcgagagattgaatcaagattatccaatatcgaacttcaactcacggaaataccgaaacataagtaatcatatcccttctcacattttttacacaggaagtaagcaagttatgttatatttccgacactataattaattcttagcagttaaaggtgctttgtctatattacatttacatacagcttgagtgatcctgaccggatatagggtcctattttcttacgtgaacggcttttcttcttgttcccgatggccttcatgtgaaaaagcactcctcgggaggcggaaaaatatcaaaagtacggggcgaagtttataatgaagatttatcgatataaattttggttatttcaggagaacaagaaagctctttacactaaaattatcagagaagaagctgatatattagccctaaggagtacaccaaaacacaagatc +RNR1_promoter: + description: placeholder + tags: [promoter] + prefix: TACA + suffix: AACT + sequence: ggatatcgtaaacaaaggcgttaccatagaaatgtactgattggcagaattactcttcaggagaatctttcatacaaaggtattccattggggaaaatctcgttaccaagtcaatgctgaactttctatggcctttgtttactatcgttaattattttacgaccacttctgggtagaaatatttcgtagccctggaacgagcttgtttacgcgttttatcccattatatggcacccaaatcaaatttaaaaagaaaaaacgcgtaaacagtgtcgggtaagttcatcctctgttactttaattgcttctttttttgaaattctaagtaaacgcgtcattttgatcctcaggacacagaaatccttgcagaatcttattgggtgttgaatagaggacgcgtaaaaacgatatggaaatttttttcatatagtgtagaaagaataggttggcgtaggtagtttcgtgtttgatagaaacctccaacaaagtctgcaactcacgttttagaataacaagtttagagtttatcttgttgcctttgttaagtcagtaccattgaataaaaattatataaaggagctaatatttcattgttggaaaattactctaccataattgaagcatatctcatccttttcatccttttcaacgcaagagagacaccaacgaacaacactttatttgttgatatattaacatcagatc +RET2_promoter: + description: placeholder + tags: [promoter] + prefix: TACA + suffix: AACT + sequence: acgatggcttcttatctcacttcaatagtactttccaccggttatacttccggcttttccctattaatacaagctacaatttcaatgggtggcaaataatgtgtagaatagaaaataagccgacagggtaataaagaaaatttttagaaaaaaaaggttagatggcttatttaagttacaggctagcgaaaaaaggaacttcagggcaagtaaagtgtttgattgggcactagcatggcttataaaggcgagcaattgtcgaaactaattaatgttgtacggactattgctgtcatctcgtggtaaatgcgtgttccaggtcgaatactacttgcacacaggcgagcggggccccataaaagtgttgccgatttgttaagttgtcttttcggtttttctactctgttattccttacttccctttttaagaactctttttatccttcatttaggatcttgcacgtttccgcctcatcacttgaattaaaacatgtctctgtcagtaaaccttggcgtttctattgttcttcatagttcaacttttattattacccgccctgcgcgtttacatttttccagcaacagccagcgaaaaattagaaaatctggttgttgacacctcaagaacaagggcaattagcctcagcgtcgaatatagatcatattagaatacctatagctccatcaaaagaaatacacaagatc +PAB1_promoter: + description: placeholder + tags: [promoter] + prefix: TACA + suffix: AACT + sequence: aaggcaagcccagaaaaatatcgcaagcacctttggtcttacagtgccaacttttggcctgccgacgttaagagtacaaagctgatggcaatgtacgacaagataacagagtctcaaaagaagtgaaacaatttttcttcaccacattttccattgttccttccccccataactataaacgtatttatgtatatatatttgcgtgtaagtgtgtgtactatagggcaccgtaaagtaataatgcttaattagttactactatgaccatataagaggtcatactgtatgaagccacaaagcagatagatcaatcatgtttaacgaaaactgttaatcgaagattatttctttttttttttctctttcctttttacaaagaaaattttttttgcgctttttgccatcaccatcgcaagttctgggacaattgttctctttcgctccagttccaaggaaagaggtttctgttttacttaatagaaagtgtcatcttgtattttatatctcttctttcttgtgtaaaattctttagttttgattttgtatttttaggacagtgagctacgaagtaacatttttacttaataaccgtttgaagcatagagcaggccctggtatcaccacctaatatctggctttttattcaataaaaactcaaaaaaaaaaatccaaaaaaaactaaaaaaccaataaaaataaaagatc +ALD6_promoter: + description: placeholder + tags: [promoter] + prefix: TACA + suffix: AACT + sequence: taagggcatgatagaattggattatgtaaaaggtgaagataccattgtagaagcaaccagcacgtcgccgtggctgatgaagtctcctcttgcccgggccgcagaaaagaggggcagtggcctgtttttcgacataaatgaggggcatggccagcaccaagacgtcattgttgcatatggcgtatccaagccgaaacggcgctcgcctcatccccacgggaataaggcagccgacaaaagaaaaacgaccgaaaaggaaccagaaagaaaaaagagggtgggcgcgccgcggacgtgtaaaaagatatgcatccagcttctatatcgctttaactttaccgttttgggcatcgggaacgtatgtaacattgatctcctcttgggaacggtgagtgcaacgaatgcgatatagcaccgaccatgtgggcaaattcgtaataaattcggggtgagggggattcaagacaagcaaccttgttagtcagctcaaacagcgatttaacggttgagtaacacatcaaaacaccgttcgaggtcaagcctggcgtgtttaacaagttcttgatatcatatataaatgtaataagaagtttggtaatattcaattcgaagtgttcagtcttttacttctcttgttttatagaagaaaaaacatcaagaaacatctttaacatacacaaacacatactatcagaatacaagatc +RPL18B_promoter: + description: placeholder + tags: [promoter] + prefix: TACA + suffix: AACT + sequence: aagaggatgtccaatattttttttaaggaataaggatacttcaagactagattcccccctgcattcccatcagaaccgtaaaccttggcgctttccttgggaagtattcaagaagtgccttgtccggtttctgtggctcacaaaccagcgcgcccgatatggctttcttttcacttatgaatgtaccagtacgggacaattagaacgctcctgtaacaatctctttgcaaatgtggggttacattctaaccatgtcacactgctgacgaaattcaaagtaaaaaaaaatgggaccacgtcttgagaacgatagattttctttattttacattgaacagtcgttgtctcagcgcgctttatgttttcattcatacttcatattataaaataacaaaagaagaatttcatattcacgcccaagaaatcaggctgctttccaaatgcaattgacacttcattagccatcacacaaaactctttcttgctggagcttcttttaaaaaagacctcagtacaccaaacacgttacccgacctcgttattttacgacaactatgataaaattctgaagaaaaaataaaaaaattttcatacttcttgcttttatttaaaccattgaatgatttcttttgaacaaaactacctgtttcaccaaaggaaatagaaagaaaaaatcaattagaagaaaacaaaaaacaaaagatc +HTB2_promoter: + description: placeholder + tags: [promoter] + prefix: TACA + suffix: AACT + sequence: tatatattaaatttgctcttgttctgtactttcctaattcttatgtaaaaagacaagaatttatgatactatttaataacaaaaaactacctaagaaaagcatcatgcagtcgaaattgaaatcgaaaagtaaaactttaacggaacatgtttgaaattctaagaaagcatacatcttcatcccttatatatagagttatgtttgatattagtagtcatgttgtaatctctggcctaagtatacgtaacgaaaatggtagcacgtcgcgtttatggcccccaggttaatgtgttctctgaaattcgcatcactttgagaaataatgggaacaccttacgcgtgagctgtgcccaccgcttcgcctaataaagcggtgttctcaaaatttctccccgttttcaggatcacgagcgccatctagttctggtaaaatcgcgcttacaagaacaaagaaaagaaacatcgcgtaatgcaacagtgagacacttgccgtcatatataaggttttggatcagtaaccgttatttgagcataacacaggtttttaaatatattattatatatcatggtatatgtgtaaaatttttttgctgactggttttgtttatttatttagctttttaaaaattttactttcttcttgttaattttttctgattgctctatactcaaaccaacaacaacttactctacaactaagatc +HHF1_promoter: + description: placeholder + tags: [promoter] + prefix: TACA + suffix: AACT + sequence: tcttggggccttaccaccagtggactttcttgctgtttgctttgttctggccattgtttgcgtttatatatttatgttagatgtttttcttattaactagaaagaaagaatataaaaggttgaggaaagagatgtatcccgaagaatacacagtcttttatatatgtatttcaacaaggagccgtggagggtactaaaaagaaaaatcgcccgggcatttcgttatcttccacgctaaaagtcaaggagagatattacggccaggatcgcaaaggtgcagagcaaggaaatgtgagaaattgtgagaacgataatgtatgggacaatgcgaaaatgtgagaacgagagcaaaaatcttttttgtatctccccgccgaatttggaaaccgcgttctgaaaacttcgcatcttcacatagtaaaactgttccgagcgcttctccccataatggttagtggtaaaaaccgaagttgtttactttagcaaatgcccgcgaatacggtggtaaattgccacccccccttccccattcattgggtaaagaccaatttgatggataaattggttgtggaaaaggtctaattctttttcctataaataccgagatattttttctatatgatggtttccgtcgcattattgtactctatagtactaaagcaacaaacaaaaacaagcaacaaatataatatagtaaaatagatc +TEF2_promoter: + description: placeholder + tags: [promoter] + prefix: TACA + suffix: AACT + sequence: ttgataggtcaagatcaatgtaaacaattactttgttatgtagagtttttttagctacctatattccaccataacatcaatcatgcggttgctggtgtatttaccaataatgtttaatgtatatatatatatatatatatggggccgtatacttacatatagtagatgtcaagcgtaggcgcttcccctgccggctgtgagggcgccataaccaaggtatctatagaccgccaatcagcaaactacctccgtacattcatgttgcacccacacatttatacacccagaccgcgacaaattacccataaggttgtttgtgacggcgtcgtacaagagaacgtgggaactttttaggctcaccaaaaaagaaagaaaaaatacgagttgctgacagaagcctcaagaaaaaaaaaattcttcttcgactatgctggaggcagagatgatcgagccggtagttaactatatatagctaaattggttccatcaccttcttttctggtgtcgctccttctagtgctatttctggcttttcctatttttttttttccatttttctttctctctttctaatatataaattctcttgcattttctatttttctctctatctattctacttgtttattcccttcaaggtttttttttaaggagtacttgtttttagaatatacggtcaacgaactataattaactaaacagatc +TEF1_promoter: + description: placeholder + tags: [promoter] + prefix: TACA + suffix: AACT + sequence: ccttgccaacagggagttcttcagagacatggaggctcaaaacgaaattattgacagcctagacatcaatagtcatacaacagaaagcgaccacccaactttggctgataatagcgtataaacaatgcatactttgtacgttcaaaatacaatgcagtagatatatttatgcatattacatataatacatatcacataggaagcaacaggcgcgttggacttttaattttcgaggaccgcgaatccttacatcacacccaatcccccacaagtgatcccccacacaccatagcttcaaaatgtttctactccttttttactcttccagattttctcggactccgcgcatcgccgtaccacttcaaaacacccaagcacagcatactaaatttcccctctttcttcctctagggtgtcgttaattacccgtactaaaggtttggaaaagaaaaaagacaccgcctcgtttctttttcttcgtcgaaaaaggcaataaaaatttttatcacgtttctttttcttgaaaatttttttttttgatttttttctctttcgatgacctcccattgatatttaagttaataaacggtcatcaatttctcaagtttcagtttcatttttcttgttctattacaactttttttacttcttgctcattagaaagaaagcatagcaatctaatctaagttttaattacaaaagatc +HHF2_promoter: + description: placeholder + tags: [promoter] + prefix: TACA + suffix: AACT + sequence: tgtggagtgtttgcttggattctttagtaaaaggggaagaacagttggaagggccaaagtggaagtcacaaaacagtggtcctatataaaagaacaagaaaaagattatttatatacaactgcggtcacaagaagcaacgcgagagagcacaacacgctgttatcacgcaaactatgttttgacaccgagccatagccgtgattgtgcgtcacattgggcgataatgaacgctaaatgaccaactcccatccgtaggagccccttagggcgtgccaatagtttcacgcgcttaatgcgaagtgctcggaacggacaactgtggtcgtttggcaccgggaaagtggtactagaccgagagtttcgcatttgtatggcaggacgttctgggagcttcgcgtctaaagctttttcgggcgcgaaatgcagaccagaccagaacaaaacaactgacaagaaggcgtttaatttaatatgttgttcactcgcgcctgggctgttgttattcggctagatacatacgtgtttgtgcgtatgtagttatatcatatataagtatattaggatgaggcggtgaaagagattttttttttttcgcttaatttattcttttctctatcttttttcctacatcttgttcaaaagagtagcaaaaacaacaatcaatacaataaaataagatc +PGK1_promoter: + description: placeholder + tags: [promoter] + prefix: TACA + suffix: AACT + sequence: gtgagtaaggaaagagtgaggaactatcgcatacctgcatttaaagatgccgatttgggcgcgaatcctttattttggcttcaccctcatactattatcagggccagaaaaaggaagtgtttccctccttcttgaattgatgttaccctcataaagcacgtggcctcttatcgagaaagaaattaccgtcgctcgtgatttgtttgcaaaaagaacaaaactgaaaaaacccagacacgctcgacttcctgtcatcctattgattgcagcttccaatttcgtcacacaacaaggtcctagcgacggctcacaggttttgtaacaagcaatcgaaggttctggaatggcgggaaagggtttagtaccacatgctatgatgcccactgtgatctccagagcaaagttcgttcgatcgtactgttactctctctctttcaaacagaattgtccgaatcgtgtgacaacaacagcctgttctcacacactcttttcttctaaccaagggggtggtttagtttagtagaacctcgtgaaacttacatttacatatatataaacttgcataaattggtcaatgcaagaaatacatatttggtcttttctaattcgtagtttttcaagttcttagatgctttctttttctcttttttacagatcatcaaggaagtaattatctactttttacaacaaatataaaacaagatc +CCW12_promoter: + description: placeholder + tags: [promoter] + prefix: TACA + suffix: AACT + sequence: cacccatgaaccacacggttagtccaaaaggggcagttcagattccagatgcgggaattagcttgctgccaccctcacctcactaacgctgcggtgtgcggatacttcatgctatttatagacgcgcgtgtcggaatcagcacgcgcaagaaccaaatgggaaaatcggaatgggtccagaactgctttgagtgctggctattggcgtctgatttccgttttgggaatcctttgccgcgcgcccctctcaaaactccgcacaagtcccagaaagcgggaaagaaataaaacgccaccaaaaaaaaaaaaataaaagccaatcctcgaagcgtgggtggtaggccctggattatcccgtacaagtatttctcaggagtaaaaaaaccgtttgttttggaatttcccatttcgcggccacctacgccgctatctttgcaacaactatctgcgataactcagcaaattttgcatattcgtgttgcagtattgcgataatgggagtcttacttccaacataacggcagaaagaaatgtgagaaaattttgcatcctttgcctccgttcaagtatataaagtcggcatgcttgataatctttctttccatcctacattgttctaattattcttattctcctttattctttcctaacataccaagaaattaatcttctgtcattcgcttaaacactatatcaataaagatc +TDH3_promoter: + description: placeholder + tags: [promoter] + prefix: TACA + suffix: AACT + sequence: cagttcgagtttatcattatcaatactgccatttcaaagaatacgtaaataattaatagtagtgattttcctaactttatttagtcaaaaaattagccttttaattctgctgtaacccgtacatgcccaaaatagggggcgggttacacagaatatataacatcgtaggtgtctgggtgaacagtttattcctggcatccactaaatataatggagcccgctttttaagctggcatccagaaaaaaaaagaatcccagcaccaaaatattgttttcttcaccaaccatcagttcataggtccattctcttagcgcaactacagagaacaggggcacaaacaggcaaaaaacgggcacaacctcaatggagtgatgcaacctgcctggagtaaatgatgacacaaggcaattgacccacgcatgtatctatctcattttcttacaccttctattaccttctgctctctctgatttggaaaaagctgaaaaaaaaggttgaaaccagttccctgaaattattcccctacttgactaataagtatataaagacggtaggtattgattgtaattctgtaaatctatttcttaaacttcttaaattctacttttatagttagtcttttttttagttttaaaacaccaagaacttagtttcgaataaacacacataaacaaacaaaagatc +SceI(Scerevisae): + description: placeholder + tags: [cds] + prefix: A + suffix: ATCC + sequence: atgaagaacatcaagaagaaccaagttatgaacttgggtccaaactctaagttgttgaaggaatacaagtctcaattgatcgaattgaacatcgaacaattcgaagctggtatcggtttgatcttgggtgacgcttacatccgatctcgcgacgaaggtaagacctactgtatgcaattcgaatggaagaacaaggcttacatggaccacgtttgtttgttgtacgaccaatgggttttgtctccaccacacaagaaggaaagagttaaccacttgggtaacttggttatcacctggggtgctcaaaccttcaagcaccaagctttcaacaagttggctaacttgttcatcgttaacaacaagaaaaccatcccaaacaacttggttgaaaactacttgaccccaatgtctttggcttactggttcatggacgacggtggtaagtgggactacaacaagaactctaccaacaagtctatcgttttgaacacccaatctttcaccttcgaagaagttgaatacttggttaagggtttgagaaacaagttccaattgaactgttacgttaagatcaacaagaacaagccaatcatctacatcgactctatgtcttacttgatcttctacaacttgatcaagccatacttgatcccacaaatgatgtacaagttgccaaacaccatctcttctgaaaccttcttgaag +mRuby2(Scerevisae): + description: placeholder + tags: [cds] + prefix: A + suffix: ATCC + sequence: atggtgtccaaaggagaggagttaatcaaggaaaacatgagaatgaaagttgtcatggagggctccgttaatggtcaccaattcaagtgtacaggggaaggtgaaggtaatccttacatgggtacacaaactatgagaattaaagtaattgaaggcggaccactaccatttgcatttgacattctggcaacgtcattcatgtacggatcacgaactttcatcaagtaccctaaaggtataccagactttttcaagcaatcttttccagagggttttacatgggaaagggttacaagatacgaagatgggggtgtcgtcacagttatgcaagatacttcattagaagatggctgccttgtctatcatgtgcaagtaagaggggtgaattttccttctaacggacctgtgatgcagaaaaagaccaaaggttgggaaccaaatactgaaatgatgtacccagctgatggaggtttgagaggctacacacacatggcgcttaaagttgatggtggaggtcatttgtcttgtagttttgttaccacttatcgttctaaaaagactgttggcaatatcaaaatgccaggaatacatgctgtagaccacagactagaaagactcgaagagagcgataacgaaatgttcgttgtacagagagagcatgccgtagccaaatttgctggcttaggcggtggtatggatgaattgtataag +Venus(Scerevisae): + description: placeholder + tags: [cds] + prefix: A + suffix: ATCC + sequence: atgtctaaaggtgaagaattattcactggtgttgtcccaattttggttgaattagatggtgatgttaatggtcacaaattttctgtctccggtgaaggtgaaggtgatgctacttacggtaaattgaccttaaaattgatttgtactactggtaaattgccagttccatggccaaccttagtcactactttaggttatggtttgcaatgttttgctagatacccagatcatatgaaacaacatgactttttcaagtctgccatgccagaaggttatgttcaagaaagaactatttttttcaaagatgacggtaactacaagaccagagctgaagtcaagtttgaaggtgataccttagttaatagaatcgaattaaaaggtattgattttaaagaaggtggtaacattttaggtcacaaattggaatacaactataactctcacaatgtttacatcactgctgacaaacaaaagaatggtatcaaagctaacttcaaaattagacacaacattgaagatggtggtgttcaattagctgaccattatcaacaaaatactccaattggtgatggtccagtcttgttaccagacaaccattacttatcctatcaatctgccttatccaaagatccaaacgaaaagagagatcacatggtcttgttagaatttgttactgctgctggtattacccatggtatggatgaattgtacaaaggatcc +mTurquoise2(Scerevisae): + description: placeholder + tags: [cds] + prefix: A + suffix: ATCC + sequence: atggtttctaaaggtgaagaattattcactggtgttgtcccaattttggttgaattagatggtgatgttaatggtcacaaattttctgtctccggtgaaggtgaaggtgatgctacttacggtaaattgaccttaaaatttatttgtactactggtaaattgccagttccatggccaaccttagtcactactttatcttggggtgttcaatgttttgcaagatacccagatcatatgaaacaacatgactttttcaagtctgccatgccagaaggttatgttcaagaaagaactatttttttcaaagatgacggtaactacaagaccagagctgaagtcaagtttgaaggtgataccttagttaatagaatcgaattaaaaggtattgattttaaagaagatggtaacattttaggtcacaaattggaatacaattatttctctgacaatgtttacatcactgctgacaaacaaaagaatggtatcaaagctaacttcaaaattagacacaacattgaagatggtggtgttcaattagctgaccattatcaacaaaatactccaattggtgatggtccagtcttgttaccagacaaccattacttatccactcaatctaagttatccaaagatccaaacgaaaagagggaccacatggtcttgttagaatttgttactgctgctggtattaccttgggtatggatgaattgtacaaa +tENO1_terminator: + description: placeholder + tags: [terminator] + prefix: ATCCTAA + suffix: CGCT + sequence: agcttttgattaagccttctagtccaaaaaacacgtttttttgtcatttatttcattttcttagaatagtttagtttattcattttatagtcacgaatgttttatgattctatatagggttgcaaacaagcatttttcattttatgttaaaacaatttcaggtttaccttttattctgcttgtggtgacgcgtgtatccgcccgctcttttggtcacccatgtat +tSSA1_terminator: + description: placeholder + tags: [terminator] + prefix: ATCCTAA + suffix: CGCT + sequence: gccaattggtgcggcaattgataataacgaaaatgtcttttaatgatctgggtataatgaggaattttccgaacgtttttactttatatatatatatacatgtaacatatattctatacgctatagagaaaggaaatttttcaattaaaaaaaaaatagagaaagagtttcacttcttgattatcgctaacactaatggttgaagtactgctactttaattttat +tADH1_terminator: + description: placeholder + tags: [terminator] + prefix: ATCCTAA + suffix: CGCT + sequence: gcgaatttcttatgatttatgatttttattattaaataagttataaaaaaaataagtgtatacaaattttaaagtgactcttaggttttaaaacgaaaattcttattcttgagtaactctttcctgtaggtcaggttgctttctcaggtatagcatgaggtcgctcttattgaccacacctctaccggcatgccgagcaaatgcctgcaaatcgctccccatttc +tPGK1_terminator: + description: placeholder + tags: [terminator] + prefix: ATCCTAA + suffix: CGCT + sequence: attgaattgaattgaaatcgatagatcaatttttttcttttctctttccccatcctttacgctaaaataatagtttattttattttttgaatattttttatttatatacgtatatatagactattatttatcttttaatgattattaagatttttattaaaaaaaaattcgctcctcttttaatgcctttatgcagtttttttttcccattcgatatttctatgt +tENO2_terminator: + description: placeholder + tags: [terminator] + prefix: ATCCTAA + suffix: CGCT + sequence: agtgcttttaactaagaattattagtcttttctgcttattttttcatcatagtttagaacactttatattaacgaatagtttatgaatctatttaggtttaaaaattgatacagttttataagttactttttcaaagactcgtgctgtctattgcataatgcactggaaggggaaaaaaaaggtgcacacgcgtggctttttcttgaatttgcagtttgaaaaat +tTDH1_terminator: + description: placeholder + tags: [terminator] + prefix: ATCCTAA + suffix: CGCT + sequence: ataaagcaatcttgatgaggataatgatttttttttgaatatacataaatactaccgtttttctgctagattttgtgatgacgtaaataagtacatattactttttaagccaagacaagattaagcattaactttacccttttctttctaagtttcaatattagttatcactgtttaaaagttatggcgagaacgtcggcggttaaaatatattaccctgaacg +URA3: + description: placeholder + tags: [selection_marker] + prefix: AAGC + suffix: ATAG + sequence: cggtttccttgaaatttttttgattcggtaatctccgaacagaaggaagaacgaaggaaggagcacagacttagattggtatatatacgcatatgtagtgttgaagaaacatgaaattgcccagtattcttaacccaactgcacagaacaaaaacgtgcaggaaacgaagataaatcatgtcgaaagctacatataaggaacgtgctgctactcatcctagtcctgttgctgccaagctatttaatatcatgcacgaaaagcaaacaaacttgtgtgcttcattggatgttcgtaccaccaaggaattactggagttagttgaagcattaggtcccaaaatttgtttactaaaaacacatgtggatatcttgactgatttttccatggagggcacagttaagccgctaaaggcattatccgccaagtacaattttttactcttcgaggacagaaaatttgctgacattggtaatacagtcaaattgcagtactctgcgggtgtatacagaatagcagaatgggcagacattacgaatgcacacggtgtggtgggcccaggtattgttagcggtttgaagcaggcggcagaagaagtaacaaaggaacctagaggccttttgatgttagcagaattgtcatgcaagggctccctatctactggagaatatactaagggtactgttgacattgcgaagagcgacaaagattttgttatcggctttattgctcaaagagacatgggtggaagagatgaaggttacgattggttgattatgacacccggtgtgggtttagatgacaagggagatgcattgggtcaacagtatagaaccgtggatgatgtggtttctacaggatctgacattattattgttggaagaggactatttgcaaagggaagggatgctaaggtagagggtgaacgttacagaaaagcaggctgggaagcatatttgagaagatgcggccagcaaaactaaaaaactgtattataagtaaatgcatgtatactaaactcacaaattagagcttcaatttaattatatcagttattaccc +LEU2: + description: placeholder + tags: [selection_marker] + prefix: AAGC + suffix: ATAG + sequence: tcgaggagaacttctagtatatctacatacctaatattattgccttattaaaaatggaatcccaacaattacatcaaaatccacattctcttcaaaatcaattgtcctgtacttccttgttcatgtgtgttcaaaaacgttatatttataggataattatactctatttctcaacaagtaattggttgtttggccgagcggtctaaggcgcctgattcaagaaatatcttgaccgcagttaactgtgggaatactcaggtatcgtaagatgcaagagttcgaatctcttagcaaccattatttttttcctcaacataacgagaacacacaggggcgctatcgcacagaatcaaattcgatgactggaaattttttgttaatttcagaggtcgcctgacgcatatacctttttcaactgaaaaattgggagaaaaaggaaaggtgagagcgccggaaccggcttttcatatagaatagagaagcgttcatgactaaatgcttgcatcacaatacttgaagttgacaatattatttaaggacctattgttttttccaataggtggttagcaatcgtcttactttctaacttttcttaccttttacatttcagcaatatatatatatatatttcaaggatataccattctaatgtctgcccctaagaagatcgtcgttttgccaggtgaccacgttggtcaagaaatcacagccgaagccattaaggttcttaaagctatttctgatgttcgttccaatgtcaagttcgatttcgaaaatcatttaattggtggtgctgctatcgatgctacaggtgttccacttccagatgaggcgctggaagcctccaagaaggctgatgccgttttgttaggtgctgtgggtggtcctaaatggggaaccggtagtgttagacctgaacaaggtttactaaaaatccgtaaagaacttcaattgtacgccaacttaagaccatgtaactttgcatccgactctcttttagacttatctccaatcaagccacaatttgctaaaggtactgacttcgttgttgtcagagaattagtgggaggtatttactttggtaagagaaaggaggacgatggtgatggtgtcgcttgggatagtgaacaatacaccgttccagaagtgcaaagaatcacaagaatggccgctttcatggccctacaacatgagccaccattgcctatttggtccttggataaagctaatgttttggcctcttcaagattatggagaaaaactgtggaggaaaccatcaagaacgaatttcctacattgaaggttcaacatcaattgattgattctgccgccatgatcctagttaagaacccaacccacctaaatggtattataatcaccagcaacatgtttggtgatatcatctccgatgaagcctccgttatcccaggttccttgggtttgttgccatctgcgtccttggcctctttgccagacaagaacaccgcatttggtttgtacgaaccatgccacggttctgctccagatttgccaaagaataaggtcaaccctatcgccactatcttgtctgctgcaatgatgttgaaattgtcattgaacttgcctgaagaaggtaaggccattgaagatgcagttaaaaaggttttggatgcaggtatcagaactggtgatttaggtggttccaacagtaccaccgaagtcggtgatgctgtcgccgaagaagttaagaaaatccttgcttaacctggaggacccttctctttagactattctactcttatgcacgtaaaaaattctaggaaatatgtattaactaggagtaaaataaccggctagtggcattcatatagccgtctgtttacatctacatcacacatttcgagtgtatatctcgcaacgttggcgttaaataggcag +HIS3: + description: placeholder + tags: [selection_marker] + prefix: AAGC + suffix: ATAG + sequence: aacacagtcctttcccgcaattttctttttctattactcttggcctcctctagtacactctatatttttttatgcctcggtaatgattttcatttttttttttccacctagcggatgactctttttttttcttagcgattggcattatcacataatgaattatacattatataaagtaatgtgatttcttcgaagaatatactaaaaaatgagcaggcaagataaacgaaggcaaagatgacagagcagaaagccctagtaaagcgtattacaaatgaaaccaagattcagattgcgatctctttaaagggtggtcccctagcgatagagcactcgatcttcccagaaaaagaggcagaagcagtagcagaacaggccacacaatcgcaagtgattaacgtccacacaggtatagggtttctggaccatatgatacatgctctggccaagcattccggctggtcgctaatcgttgagtgcattggtgacttacacatagacgaccatcacaccactgaggactgcgggattgctctcggtcaagcttttaaagaggccctaggggccgtgcgtggagtaaaaaggtttggatcaggatttgcgcctttggatgaggcactttccagagcggtggttgatctttcgaacaggccgtacgcagttgtcgaacttggtttgcaaagggagaaagtaggtgatctctcttgcgagatgatcccgcattttcttgaaagctttgcagaggctagcagaattaccctccacgttgattgtctgcgaggcaagaatgatcatcaccgtagtgagagtgcgttcaaggctcttgcggttgccataagagaagccacctcgcccaatggtaccaacgatgttccctccaccaaaggtgttcttatgtagtgacaccgattatttaaagctgctgcatacgatatatatacatgtgtatatatgtatacctatgaatgtcagtaagtatgtatacgaacagtatgatactgaagatgacaaggtaatgcatcattctatacgtgtcattctgaacgaggcgcgctttccttttttctttttgctttttctttttttttctcttgaactcgacggatcata +KanamycinR: + description: placeholder + tags: [selection_marker] + prefix: AAGC + suffix: ATAG + sequence: agcttgcctcgtccccgccgggtcacccggccagcgacatggaggcccagaataccctccttgacagtcttgacgtgcgcagctcaggggcatgatgtgactgtcgcccgtacatttagcccatacatccccatgtataatcatttgcatccatacattttgatggccgcacggcgcgaagcaaaaattacggctcctcgctccagacctgcgagcagggaaacgctcccctcacagacgcgttgaattgtccccacgccgcgcccctgtagagaaatataaaaggttaggatttgccactgaggttcttctttcatatacttccttttaaaatcttgctaggatacagttctcacatcacatccgaacataaacaaaaatgggtaaggaaaagactcacgtttcgaggccgcgattaaattccaacatggatgctgatttatatgggtataaatgggctcgcgataatgtcgggcaatcaggtgcgacaatctatcgattgtatgggaagcccgatgcgccagagttgtttctgaaacatggcaaaggtagcgttgccaatgatgttacagatgagatggtcagactaaactggctgacggaatttatgcctcttccgaccatcaagcattttatccgtactcctgatgatgcatggttactcaccactgcgatccccggcaaaacagcattccaggtattagaagaatatcctgattcaggtgaaaatattgttgatgcgctggcagtgttcctgcgccggttgcattcgattcctgtttgtaattgtccttttaacagcgatcgcgtatttcgtctggctcaggcgcaatcacgaatgaataacggtttggttgatgcgagtgattttgatgacgagcgtaatggctggcctgttgaacaagtctggaaagaaatgcataagcttttgccattctcaccggattcagtcgtcactcatggtgatttctcacttgataaccttatttttgacgaggggaaattaataggttgtattgatgttggacgagtcggaatcgcagaccgataccaggatcttgccatcctatggaactgcctcggtgagttttctccttcattacagaaacggctttttcaaaaatatggtattgataatcctgatatgaataaattgcagtttcatttgatgctcgatgagtttttctaaagtaactgacaataaaaagattcttgttttcaagaacttgtcatttgtatagtttttttatattgtagttgttctattttaatcaaatgttagcgtgatttatattttttttcgcctcgacatcatctgcccagatgcgaagttaagtgcgcagaaagtaatatcatgcgtcaatcgtatgtgaatgctggtcgctatactg +NourseothricinR: + description: placeholder + tags: [selection_marker] + prefix: AAGC + suffix: ATAG + sequence: agcttgcctcgtccccgccgggtcacccggccagcgacatggaggcccagaataccctccttgacagtcttgacgtgcgcagctcaggggcatgatgtgactgtcgcccgtacatttagcccatacatccccatgtataatcatttgcatccatacattttgatggccgcacggcgcgaagcaaaaattacggctcctcgctccagacctgcgagcagggaaacgctcccctcacagacgcgttgaattgtccccacgccgcgcccctgtagagaaatataaaaggttaggatttgccactgaggttcttctttcatatacttccttttaaaatcttgctaggatacagttctcacatcacatccgaacataaacaaaaatgggtactaccttagatgatacagcctacagatacagaacatcagtccctggtgatgctgaagcaattgaggctttagacggttcattcaccaccgacaccgtctttagagtaaccgccaccggtgatggatttaccttaagagaagtcccagtcgaccctccattaactaaagtctttccagatgatgaatctgatgacgaaagcgacgacggagaagatggtgacccagattcaagaactttcgtagcatacggtgatgacggtgatttggctggttttgtagtcgtttcttattcaggttggaatagaaggttgaccgttgaagatatagaagtcgccccagagcatagaggtcatggtgtaggaagagctttgatgggtttggctacagaatttgcaagagagagaggagccggtcatttatggttagaagttactaatgttaacgcccctgctatccatgcttatagaagaatgggtttcacattatgtggtttagatactgctttatatgatggaacagcatctgacggtgaacaggccttgtatatgtctatgccttgcccttaaagtaactgacaataaaaagattcttgttttcaagaacttgtcatttgtatagtttttttatattgtagttgttctattttaatcaaatgttagcgtgatttatattttttttcgcctcgacatcatctgcccagatgcgaagttaagtgcgcagaaagtaatatcatgcgtcaatcgtatgtgaatgctggtcgctatactg +HygromycinR: + description: placeholder + tags: [selection_marker] + prefix: AAGC + suffix: ATAG + sequence: agcttgcctcgtccccgccgggtcacccggccagcgacatggaggcccagaataccctccttgacagtcttgacgtgcgcagctcaggggcatgatgtgactgtcgcccgtacatttagcccatacatccccatgtataatcatttgcatccatacattttgatggccgcacggcgcgaagcaaaaattacggctcctcgctccagacctgcgagcagggaaacgctcccctcacagacgcgttgaattgtccccacgccgcgcccctgtagagaaatataaaaggttaggatttgccactgaggttcttctttcatatacttccttttaaaatcttgctaggatacagttctcacatcacatccgaacataaacaaaaatgggtaaaaagcctgaactcaccgcgacgtctgtcgagaagtttctgatcgaaaagttcgacagcgtgtccgacctgatgcagctctcggagggcgaagaatctcgtgctttcagcttcgatgtaggagggcgtggatatgtcctgcgggtaaatagctgcgccgatggtttctacaaagatcgttatgtttatcggcactttgcatcggccgcgctcccgattccggaagtgcttgacattggggaatttagcgagagcctgacctattgcatctcccgccgtgcacagggtgtcacgttgcaagacctgcctgaaaccgaactgcccgctgttctgcaaccggtcgcggaggccatggatgcgatcgctgcggccgatcttagccagacgagcgggttcggcccattcggaccgcaaggaatcggtcaatacactacatggcgtgatttcatatgcgcgattgctgatccccatgtgtatcactggcaaactgtgatggacgacaccgtcagtgcgtccgtcgcgcaggctctcgatgagctgatgctttgggccgaggactgccccgaagtccggcacctcgtgcacgcggatttcggctccaacaatgtcctgacggacaatggccgcataacagcggtcattgactggagcgaggcgatgttcggggattcccaatacgaggtcgccaacatcttcttctggaggccgtggttggcttgtatggagcagcagacgcgctacttcgagcggaggcatccggagcttgcaggatcgccgcggctccgggcgtatatgctccgcattggtcttgaccaactctatcagagcttggttgacggcaatttcgatgatgcagcttgggcgcagggtcgatgcgacgcaatcgtccgatccggagccgggactgtcgggcgtacacaaatcgcccgcagaagcgcggccgtctggaccgatggctgtgtagaagtactcgccgatagtggaaaccgacgccccagcactcgtccgagggcaaaggaataaagtaactgacaataaaaagattcttgttttcaagaacttgtcatttgtatagtttttttatattgtagttgttctattttaatcaaatgttagcgtgatttatattttttttcgcctcgacatcatctgcccagatgcgaagttaagtgcgcagaaagtaatatcatgcgtcaatcgtatgtgaatgctggtcgctatactg +ZeocinR: + description: placeholder + tags: [selection_marker] + prefix: AAGC + suffix: ATAG + sequence: agcttgcctcgtccccgccgggtcacccggccagcgacatggaggcccagaataccctccttgacagtcttgacgtgcgcagctcaggggcatgatgtgactgtcgcccgtacatttagcccatacatccccatgtataatcatttgcatccatacattttgatggccgcacggcgcgaagcaaaaattacggctcctcgctccagacctgcgagcagggaaacgctcccctcacagacgcgttgaattgtccccacgccgcgcccctgtagagaaatataaaaggttaggatttgccactgaggttcttctttcatatacttccttttaaaatcttgctaggatacagttctcacatcacatccgaacataaacaaaaatggctaaattaacatctgccgttcctgttttaacagctagggatgttgcaggtgctgtagagttttggacagataggttaggattctcaagagactttgttgaggacgattttgctggtgttgtcagggatgacgttactttatttatctcagcagtccaagatcaagttgtccctgataatacattggcttgggtctgggtcaggggtttagatgaattatatgctgaatggtcagaagttgtatctacaaacttcagagatgcttctggtccagctatgaccgagattggtgaacagccatggggtagagaatttgctttgagagatccagctggaaattgtgttcattttgttgctgaagaacaagattaaagtaactgacaataaaaagattcttgttttcaagaacttgtcatttgtatagtttttttatattgtagttgttctattttaatcaaatgttagcgtgatttatattttttttcgcctcgacatcatctgcccagatgcgaagttaagtgcgcagaaagtaatatcatgcgtcaatcgtatgtgaatgctggtcgctatactg +2micron: + description: placeholder + tags: [origin] + prefix: GTCT + suffix: AAGC + sequence: aacgaagcatctgtgcttcattttgtagaacaaaaatgcaacgcgagagcgctaatttttcaaacaaagaatctgagctgcatttttacagaacagaaatgcaacgcgaaagcgctattttaccaacgaagaatctgtgcttcatttttgtaaaacaaaaatgcaacgcgagagcgctaatttttcaaacaaagaatctgagctgcatttttacagaacagaaatgcaacgcgagagcgctattttaccaacaaagaatctatacttcttttttgttctacaaaaatgcatcccgagagcgctatttttctaacaaagcatcttagattactttttttctcctttgtgcgctctataatgcagtctcttgataactttttgcactgtaggtccgttaaggttagaagaaggctactttggtgtctattttctcttccataaaaaaagcctgactccacttcccgcgtttactgattactagcgaagctgcgggtgcattttttcaagataaaggcatccccgattatattctataccgatgtggattgcgcatactttgtgaacagaaagtgatagcgttgatgattcttcattggtcagaaaattatgaacggtttcttctattttgtctctatatactacgtataggaaatgtttacattttcgtattgttttcgattcactctatgaatagttcttactacaatttttttgtctaaagagtaatactagagataaacataaaaaatgtagaggtcgagtttagatgcaagttcaaggagcgaaaggtggatgggtaggttatatagggatatagcacagagatatatagcaaagagatacttttgagcaatgtttgtggaagcggtattcgcaatattttagtagctcgttacagtccggtgcgtttttggttttttgaaagtgcgtcatcagagcgcttttggttttcaaaagcgctctgaagttcctatactttctagctagagaataggaacttc +CEN_ARS: + description: placeholder + tags: [origin] + prefix: GTCT + suffix: AAGC + sequence: atcacgtgctataaaaataattataatttaaattttttaatataaatatataaattaaaaatagaaagtaaaaaaagaaattaaagaaaaaatagtttttgttttccgaagatgtaaaagactctagggggatcgccaacaaatactaccttttatcttgctcttcctgctctcaggtattaatgccgaattgtttcatcttgtctgtgtagaaaaccacacacgaaaatcctgtgattttacattttacttatcgttaatcgaatgtatatctatttaatctgcttttcttgtctaataaatatatatgtaaagtacgctttttgttgaaattttttaaacctttgtttatttttttttcttcattccgtaactcttctaccttctttatttactttctaaaatccaaatacaaaacataaaaataaataaacacagagtaaattcccaaattattccatcattaaaagatacgaggcgcgtgtaagttacaggcaagcgatccgtc diff --git a/parts/primers.tsv b/parts/primers.tsv new file mode 100644 index 0000000..b5752d6 --- /dev/null +++ b/parts/primers.tsv @@ -0,0 +1,166 @@ +id primer +P1 AAACACGTGGCAAACATTCC +P2 AAACCGGAGCCATACAGTAC +P3 AAAGCACTCTTAGGCCTCTG +P4 AAAGGGGCCGTCAATATCAG +P5 AAATAAGACGACGACCCTCG +P6 AACGATGATGCTCACTCTCG +P7 AAGAATTACTGACCCCTCGG +P8 AAGACGATCCGAGCCATTAC +P9 AAGGAACTATGGCATCGAGC +P10 AAGGACTGCATACCAGGTTG +P11 AAGGATATGTAGACACCGCC +P12 AAGGCCCAGAAGGATACAAC +P13 AAGGCGCTCGGATAATACTC +P14 AAGGTATGTATAGCGACCGC +P15 AATAGGAACCTCTTACGCGG +P16 AATATCACGCAAAAGCACCG +P17 AATCAGTTTCTTTGGCAGCC +P18 AATGCAAAGCTATTAGCGCG +P19 AATGCGTCATTTTACACGGC +P20 AATGTCCTTAGGCAGTCGTC +P21 ACAACGAGCAGACCGAATAG +P22 ACAAGGAGTCGGCATATCAC +P23 ACAGAACGAACAGGCACTAC +P24 ACAGGAAGCAAGGTATACGC +P25 ACAGGGTATATTGAGTGCCC +P26 ACATAAGCGATCCCAAGGTC +P27 ACATCGCATACCAGAACAGG +P28 ACATTAAATTTCGCCGTGGC +P29 ACCACAGGTCAAGATTCACG +P30 ACCCGTATCGCATAAGGATG +P31 ACGAGATGATGCACCGATAG +P32 ACGATGGGGACATAGAACAC +P33 ACGGAGCCCTTATTGTAACC +P34 ACGTATGGGGAACACTACAC +P35 ACGTGAAACTGTATCGAGCC +P36 ACGTTCAGTTTTCCAATGGC +P37 ACTAGATTAGCAAGGCACCC +P38 ACTGGACCCAATAAAAGGCC +P39 ACTTCGATTGGCAAGGACTG +P40 AGAACATAGCATTCACGGGG +P41 AGACAACAATCTGAGGCTGG +P42 AGACAAGCCTTAACCGTAGG +P43 AGACACAAGGCTGATTCCAG +P44 AGACATGGGATTGACCACAC +P45 AGAGAGGCATGATTGACCTC +P46 AGAGTTGCACCTAGAATCCG +P47 AGATAGATGCTCCGTCAAGC +P48 AGATAGTCACGCACAAGACC +P49 AGATTAGCCGACTTTCCTGG +P50 AGATTAGCTGCCGATACTGG +P51 AGATTGTTACTCCGACGGAC +P52 AGATTTCCGACGAGATTCCC +P53 AGCATCCGTCTAAATCTCGG +P54 AGCTATAAGAATTGCCGGGC +P55 AGCTATGATCCCGGTGTAAC +P56 AGCTCAATCTAACAGTGGGG +P57 AGGACACCAGACCAATGAAG +P58 AGGGCTAATTACCATCAGCG +P59 AGGTGATCTGACGAATGTCC +P60 AGTAAAGCATAGTGCCCAGC +P61 AGTAGTATCCGAATCGCTGC +P62 AGTATCTCAGCAAGGGCAAC +P63 AGTATTAGGCGTCAAGGTCC +P64 AGTATTCTTACAGCCAGCCG +P65 AGTATTGCCGGACTAAACCC +P66 AGTCCCAAGTTCAGACGTAC +P67 AGTCCGACACAATGTGACAC +P68 AGTGAACTGACCGAATCCTC +P69 AGTGGTCTGTAAACCGTACC +P70 AGTGTTTTCCATTTTCCGCG +P71 AGTTATAAGGGTCCGATGCC +P72 AGTTGCAGTATCTAACCCGC +P73 AGTTGTAATATCACCCGCGC +P74 ATACGTGGCTAGCATGAGAC +P75 ATACTGTAAGAACCACGCGG +P76 ATAGATCATGTCGGCAGTCG +P77 ATAGATGGTGCCTACATGCG +P78 ATCACAACAAAGGACGGGTC +P79 ATCAGACAACACAGAGGCTG +P80 ATCCAGGAGGTCTAGGAACC +P81 ATCCTAGAAAAGGCGAAGGC +P82 ATGCCATGACGACAACTAGC +P83 ATGCTAGCTGGAACTATCGG +P84 ATTAGGATTGCGAGCGACAC +P85 ATTAGTACACTCCGTGAGCG +P86 ATTCAAGGGTTGGACGACTC +P87 ATTCTCACGACGCAAGATGG +P88 ATTGACGGGAACTACACTCG +P89 CACTCGATAGGTACAACCGG +P90 CAGACCTACGGATCTTAGCG +P91 CCACGAGATAAGAGGATGGC +P92 CCAGAGCTTAGGGGACATAC +P93 CCCGAGGGGAGAAATATACC +P94 CCGAGGGAACCATGATACAG +P95 CCGGGAGGAAGATATAGCAC +P96 CCGGTTGTACCTATCGAGTG +P97 CCGTGCGACAAGATTTCAAG +P98 CCTTTAACAGGACATGCAGC +P99 CGAACGCAAAAGTCCTCAAG +P100 CGATAGAACGACCAGGTAGC +P101 CGGATCGAACTTAGGTAGCC +P102 CGGGAGGAAGTCTTTAGACC +P103 CTAATATCCCTGAGCGACGG +P104 CTAGGGAACCAGGCTTAACG +P105 CTAGGGGATGGTCCAATACG +P106 CTATAGAATCCGGGCTGGTC +P107 CTGCTAGGGGCTACTTATCG +P108 GAAAAGTCCCAATGAGTGCC +P109 GAAGTGGTTTGCCTAAACGC +P110 GACCATGCAAGGAGAGGTAC +P111 GATACATAGACTTGGCCCCG +P112 GCACGCAAAAGGACATAACC +P113 GCAGCGTTTTAGCCTACAAG +P114 GCATAAAGTTGACAGGCCAG +P115 GCTAAATAGAGGGAAGCCCC +P116 GGAAAACTAAGACAAGGCGC +P117 GGAAACAATAACCATCGGCG +P118 GGGCACCGATTAAGAAATGC +P119 GGGTTGTCTCCTCTGATAGC +P120 GTACTCAGAGATTGCCGGAG +P121 GTATAAGATCAGCCGGACCC +P122 GTATGTCGGCTCTCGTATCG +P123 GTTCAGAGGTACGAACCCTC +P124 GTTGCATCTAAGCCAAGTGC +P125 TAAAGAGAGGGCGTCCAATC +P126 TAACGACGTGCCGAACTTAG +P127 TAAGATAGCACCACGGATGG +P128 TAAGGATTCATCAGGTGCGC +P129 TAAGGGACGATGCTTAACCC +P130 TACCACGAAATGCACAGGAG +P131 TACTGATAATTCGGACGCCC +P132 TACTTGAATACCACGTGGCC +P133 TAGCCAGGCAAAAGAGATCC +P134 TAGCTCGATAATCAAGGGGC +P135 TAGTGACCTAATGCCATGGG +P136 TAGTTGAGAACACGAACCCG +P137 TATAACAGGCTGCTGAGACC +P138 TATACTGAAGAACGGCCCAG +P139 TATCAATCCGGAACCAGTGC +P140 TATCACGGAAGGACTCAACG +P141 TCAAAGGAGCACGAACCTAC +P142 TCAAGGTCCGTTATGGAACC +P143 TCACATAGAAGGACATGGCG +P144 TCACTTGGTATCGAGAACGG +P145 TCAGCCTTTCATTGATTGCG +P146 TCATCGACAAGATACAGGCG +P147 TCCAATTATACGGAGCAGGC +P148 TCGAATATGCTGTAACCCCG +P149 TCGACCAGGTTATCATGAGC +P150 TCGAGACAAGAACGATTCCC +P151 TCTAGGACTATCACCGGAGG +P152 TCTTCATAAGCCAGAGTGCC +P153 TCTTGCGATAGACACAAGCC +P154 TGAGCCATAAAAGCAAAGCG +P155 TGAGCGCAGAACTATCAGAC +P156 TGCATAGTATCCCAACAGGG +P157 TGCCAAAGGGTAGAGACATC +P158 TGCTGAATGAGAAACCTCGG +P159 TGGGGACGACTTATAATGCC +P160 TGTGGACCCTATCAAACGAG +P161 TTAGCTCAGGTCCAAAGTCC +P162 TTAGTAGGCAAGCATACCCG +P163 TTCGGGAGCGGATTATACAC +P164 TTCTGGGACTGGATAACACG +P165 TTGACAGACAATCCGTAGGC From ad7edc5ff8c2627c6318e192f42c9854537b87a3 Mon Sep 17 00:00:00 2001 From: Keoni Gandall Date: Mon, 14 Oct 2024 09:21:56 -0700 Subject: [PATCH 2/9] yeast toolkit update cds parts --- parts/parts/yeast_toolkit.yaml | 8 ++++---- 1 file changed, 4 insertions(+), 4 deletions(-) diff --git a/parts/parts/yeast_toolkit.yaml b/parts/parts/yeast_toolkit.yaml index dbcf2ff..5f9686d 100644 --- a/parts/parts/yeast_toolkit.yaml +++ b/parts/parts/yeast_toolkit.yaml @@ -140,25 +140,25 @@ SceI(Scerevisae): description: placeholder tags: [cds] prefix: A - suffix: ATCC + suffix: GGATCC sequence: atgaagaacatcaagaagaaccaagttatgaacttgggtccaaactctaagttgttgaaggaatacaagtctcaattgatcgaattgaacatcgaacaattcgaagctggtatcggtttgatcttgggtgacgcttacatccgatctcgcgacgaaggtaagacctactgtatgcaattcgaatggaagaacaaggcttacatggaccacgtttgtttgttgtacgaccaatgggttttgtctccaccacacaagaaggaaagagttaaccacttgggtaacttggttatcacctggggtgctcaaaccttcaagcaccaagctttcaacaagttggctaacttgttcatcgttaacaacaagaaaaccatcccaaacaacttggttgaaaactacttgaccccaatgtctttggcttactggttcatggacgacggtggtaagtgggactacaacaagaactctaccaacaagtctatcgttttgaacacccaatctttcaccttcgaagaagttgaatacttggttaagggtttgagaaacaagttccaattgaactgttacgttaagatcaacaagaacaagccaatcatctacatcgactctatgtcttacttgatcttctacaacttgatcaagccatacttgatcccacaaatgatgtacaagttgccaaacaccatctcttctgaaaccttcttgaag mRuby2(Scerevisae): description: placeholder tags: [cds] prefix: A - suffix: ATCC + suffix: GGATCC sequence: atggtgtccaaaggagaggagttaatcaaggaaaacatgagaatgaaagttgtcatggagggctccgttaatggtcaccaattcaagtgtacaggggaaggtgaaggtaatccttacatgggtacacaaactatgagaattaaagtaattgaaggcggaccactaccatttgcatttgacattctggcaacgtcattcatgtacggatcacgaactttcatcaagtaccctaaaggtataccagactttttcaagcaatcttttccagagggttttacatgggaaagggttacaagatacgaagatgggggtgtcgtcacagttatgcaagatacttcattagaagatggctgccttgtctatcatgtgcaagtaagaggggtgaattttccttctaacggacctgtgatgcagaaaaagaccaaaggttgggaaccaaatactgaaatgatgtacccagctgatggaggtttgagaggctacacacacatggcgcttaaagttgatggtggaggtcatttgtcttgtagttttgttaccacttatcgttctaaaaagactgttggcaatatcaaaatgccaggaatacatgctgtagaccacagactagaaagactcgaagagagcgataacgaaatgttcgttgtacagagagagcatgccgtagccaaatttgctggcttaggcggtggtatggatgaattgtataag Venus(Scerevisae): description: placeholder tags: [cds] prefix: A - suffix: ATCC + suffix: GGATCC sequence: atgtctaaaggtgaagaattattcactggtgttgtcccaattttggttgaattagatggtgatgttaatggtcacaaattttctgtctccggtgaaggtgaaggtgatgctacttacggtaaattgaccttaaaattgatttgtactactggtaaattgccagttccatggccaaccttagtcactactttaggttatggtttgcaatgttttgctagatacccagatcatatgaaacaacatgactttttcaagtctgccatgccagaaggttatgttcaagaaagaactatttttttcaaagatgacggtaactacaagaccagagctgaagtcaagtttgaaggtgataccttagttaatagaatcgaattaaaaggtattgattttaaagaaggtggtaacattttaggtcacaaattggaatacaactataactctcacaatgtttacatcactgctgacaaacaaaagaatggtatcaaagctaacttcaaaattagacacaacattgaagatggtggtgttcaattagctgaccattatcaacaaaatactccaattggtgatggtccagtcttgttaccagacaaccattacttatcctatcaatctgccttatccaaagatccaaacgaaaagagagatcacatggtcttgttagaatttgttactgctgctggtattacccatggtatggatgaattgtacaaaggatcc mTurquoise2(Scerevisae): description: placeholder tags: [cds] prefix: A - suffix: ATCC + suffix: GGATCC sequence: atggtttctaaaggtgaagaattattcactggtgttgtcccaattttggttgaattagatggtgatgttaatggtcacaaattttctgtctccggtgaaggtgaaggtgatgctacttacggtaaattgaccttaaaatttatttgtactactggtaaattgccagttccatggccaaccttagtcactactttatcttggggtgttcaatgttttgcaagatacccagatcatatgaaacaacatgactttttcaagtctgccatgccagaaggttatgttcaagaaagaactatttttttcaaagatgacggtaactacaagaccagagctgaagtcaagtttgaaggtgataccttagttaatagaatcgaattaaaaggtattgattttaaagaagatggtaacattttaggtcacaaattggaatacaattatttctctgacaatgtttacatcactgctgacaaacaaaagaatggtatcaaagctaacttcaaaattagacacaacattgaagatggtggtgttcaattagctgaccattatcaacaaaatactccaattggtgatggtccagtcttgttaccagacaaccattacttatccactcaatctaagttatccaaagatccaaacgaaaagagggaccacatggtcttgttagaatttgttactgctgctggtattaccttgggtatggatgaattgtacaaa tENO1_terminator: description: placeholder From 8eb9558bec05bf709287b9ad46c879b6d2d67ca5 Mon Sep 17 00:00:00 2001 From: Keoni Gandall Date: Mon, 14 Oct 2024 09:46:57 -0700 Subject: [PATCH 3/9] add ctag --- parts/parts/yeast_toolkit.yaml | 36 ++++++++++++++++++++++++++++++++++ 1 file changed, 36 insertions(+) diff --git a/parts/parts/yeast_toolkit.yaml b/parts/parts/yeast_toolkit.yaml index 5f9686d..38f0060 100644 --- a/parts/parts/yeast_toolkit.yaml +++ b/parts/parts/yeast_toolkit.yaml @@ -196,6 +196,42 @@ tTDH1_terminator: prefix: ATCCTAA suffix: CGCT sequence: ataaagcaatcttgatgaggataatgatttttttttgaatatacataaatactaccgtttttctgctagattttgtgatgacgtaaataagtacatattactttttaagccaagacaagattaagcattaactttacccttttctttctaagtttcaatattagttatcactgtttaaaagttatggcgagaacgtcggcggttaaaatatattaccctgaacg +tENO1_terminator_ctag: + description: placeholder + tags: [terminator_ctag] + prefix: ATGT + suffix: CGCT + sequence: agcttttgattaagccttctagtccaaaaaacacgtttttttgtcatttatttcattttcttagaatagtttagtttattcattttatagtcacgaatgttttatgattctatatagggttgcaaacaagcatttttcattttatgttaaaacaatttcaggtttaccttttattctgcttgtggtgacgcgtgtatccgcccgctcttttggtcacccatgtat +tSSA1_terminator_ctag: + description: placeholder + tags: [terminator_ctag] + prefix: ATGT + suffix: CGCT + sequence: gccaattggtgcggcaattgataataacgaaaatgtcttttaatgatctgggtataatgaggaattttccgaacgtttttactttatatatatatatacatgtaacatatattctatacgctatagagaaaggaaatttttcaattaaaaaaaaaatagagaaagagtttcacttcttgattatcgctaacactaatggttgaagtactgctactttaattttat +tADH1_terminator_ctag: + description: placeholder + tags: [terminator_ctag] + prefix: ATGT + suffix: CGCT + sequence: gcgaatttcttatgatttatgatttttattattaaataagttataaaaaaaataagtgtatacaaattttaaagtgactcttaggttttaaaacgaaaattcttattcttgagtaactctttcctgtaggtcaggttgctttctcaggtatagcatgaggtcgctcttattgaccacacctctaccggcatgccgagcaaatgcctgcaaatcgctccccatttc +tPGK1_terminator_ctag: + description: placeholder + tags: [terminator_ctag] + prefix: ATGT + suffix: CGCT + sequence: attgaattgaattgaaatcgatagatcaatttttttcttttctctttccccatcctttacgctaaaataatagtttattttattttttgaatattttttatttatatacgtatatatagactattatttatcttttaatgattattaagatttttattaaaaaaaaattcgctcctcttttaatgcctttatgcagtttttttttcccattcgatatttctatgt +tENO2_terminator_ctag: + description: placeholder + tags: [terminator_ctag] + prefix: ATGT + suffix: CGCT + sequence: agtgcttttaactaagaattattagtcttttctgcttattttttcatcatagtttagaacactttatattaacgaatagtttatgaatctatttaggtttaaaaattgatacagttttataagttactttttcaaagactcgtgctgtctattgcataatgcactggaaggggaaaaaaaaggtgcacacgcgtggctttttcttgaatttgcagtttgaaaaat +tTDH1_terminator_ctag: + description: placeholder + tags: [terminator_ctag] + prefix: ATGT + suffix: CGCT + sequence: ataaagcaatcttgatgaggataatgatttttttttgaatatacataaatactaccgtttttctgctagattttgtgatgacgtaaataagtacatattactttttaagccaagacaagattaagcattaactttacccttttctttctaagtttcaatattagttatcactgtttaaaagttatggcgagaacgtcggcggttaaaatatattaccctgaacg URA3: description: placeholder tags: [selection_marker] From ce5545f8a3d0c72b8ded1fbf00b9cfb1fbfaddd7 Mon Sep 17 00:00:00 2001 From: Keoni Gandall Date: Tue, 15 Oct 2024 15:42:15 -0700 Subject: [PATCH 4/9] parts --- parts/linkers.md | 15 +- parts/parts/bsub.yaml | 6 - parts/parts/ecoli/ecoli.yaml | 103 ----------- parts/parts/ecoli/origins.yaml | 52 ------ parts/parts/ecoli/promoters.yaml | 101 ----------- parts/parts/vnat.yaml | 290 ++++++++++++++++++++++++------- 6 files changed, 232 insertions(+), 335 deletions(-) delete mode 100644 parts/parts/ecoli/ecoli.yaml delete mode 100644 parts/parts/ecoli/origins.yaml delete mode 100644 parts/parts/ecoli/promoters.yaml diff --git a/parts/linkers.md b/parts/linkers.md index 4cb98b4..1bb5f8b 100644 --- a/parts/linkers.md +++ b/parts/linkers.md @@ -109,8 +109,8 @@ Often, users will want to move transcription units to new organisms of interest. * [C] AATG - ATCC (cds) * [T] ATCC - CGCT (terminator) * [B] CGCT - GTCT (linker suffix) -* [D] GTCT - AAGC (target origin of replication) -* [S] AAGC - ATAG (target selective marker) +* [S] GTCT - AAGC (target selective marker) +* [D] AAGC - ATAG (target origin of replication) * [E2] ATAG - CGAG (e coli vector 2) ``` @@ -125,10 +125,10 @@ Instead of shuttle vectors, users will sometimes want to integrate sections of D * [P] TACA - AACT (promoter) * [R] AACT - AATG (rbs) * [C] AATG - ATCC (cds) -* [T] ATCC - GTCT (terminator) -* [B] GTCT - ATAG (linker suffix) -* [D] GTCT - AAGC (target ori / downstream homology) -* [S] AAGC - ATAG (target selective marker) +* [T] ATCC - CGCT (terminator) +* [B] CGCT - GTCT (linker suffix) +* [S] GTCT - AAGC (target selective marker) +* [D] AAGC - ATAG (target upstream homology) * [EC3] ATAG - ATTA (e coli vector 3) * [U] ATTA - CGAG (upstream homology) ``` @@ -240,5 +240,4 @@ This collection has 384 linkers. There are 96 linkers for building independent t Each overhang is assigned a number. The reverse complement of each overhang is represented as the negative version of its number, which is also how we are able to flip constructs. -Linkers are named with a 1 letter + 3 number scheme, separated by underscores ( _ ) in the format `Y_X_X_X`. Y describes the direction (F, or forward, for prefix, and R, or reverse, for suffix) of the linker. The first 2 numbers describe the two overhangs which the linker itself will be cut out with, the third number describes the overhang which the linker will introduce to the construct. After a GoldenGate assembly and transformation, anything between the prefix and suffix lin -kers can be cut out with BsaI and used in another assembly. +Linkers are named with a 1 letter + 3 number scheme, separated by underscores ( _ ) in the format `Y_X_X_X`. Y describes the direction (F, or forward, for prefix, and R, or reverse, for suffix) of the linker. The first 2 numbers describe the two overhangs which the linker itself will be cut out with, the third number describes the overhang which the linker will introduce to the construct. After a GoldenGate assembly and transformation, anything between the prefix and suffix linkers can be cut out with BsaI and used in another assembly. diff --git a/parts/parts/bsub.yaml b/parts/parts/bsub.yaml index 6b3bdfa..291dc0d 100644 --- a/parts/parts/bsub.yaml +++ b/parts/parts/bsub.yaml @@ -100,12 +100,6 @@ PxylA_xylR: suffix: AACT tags: [promoter] sequence: AGCGGATAACAATTTCACACAGGAGGCCGATCGCGTTATGCAGCAATGGCAAGAACGTCCCGGGGAGCTCCTAACTTATAGGGGTAACACTTAAAAAAGAATCAATAACGATAGAAACCGCTCCTAAAGCAGGTGCATTTTTTCCTAACGAAGAAGGCAATAGTTCACATTTATTGTCTAAATGAGAATGGACTCTAGAAGAAACTTCGTTTTTAATCGTATTTAAAACAATGGGATGAGATTCAATTATATGATTTCTCAAGATAACAGCTTCTATATCAAATGTATTAAGGATATTGGTTAATCCAATTCCGATATAAAAGCCAAAGTTTTGAAGTGCATTTAACATTTCTACATCATTTTTATTTGCGCGTTCCACAATCTCTTTTCGAGAAATATTCTTTTCTTCTTTAGAGAGCGAAGCCAGTAACGCTTTTTCAGAAGCATATAATTCCCAACAGCCTCGATTTCCACAGCTGCATTTGGGTCCATTAAAATCTATCGTCATATGACCCATTTCCCCAGAAAAACCCTGAACACCTTTATACAATTCGTTGTTAATAACAAGTCCAGTTCCAATTCCGATATTAATACTGATGTAAACGATGTTTTCATAGTTTTTTGTCATACCAAATACTTTTTCACCGTATGCTCCTGCATTAGCTTCATTTTCAACAAAAACCGGAACATTAAACTCACTCTCAATTAAAAACTGCAAATCTTTGATATTCCAATTTAAGTTAGGCATGAAAATAATTTGCTGATGACGATCTACAAGGCCTGGAACACAAATTCCTATTCCGACTAGACCATAAGGGGACTCAGGCATATGGGTTACAAAACCATGAATAAGTGCAAATAAAATCTCTTTTACTTCACTAGCGGAAGAACTAGACAAGTCAGAAGTCTTCTCGAGAATAATATTTCCTTCTAAGTCGGTTAGAATTCCGTTAAGATAGTCGACTCCTATATCAATACCAATCGAGTAGCCTGCATTCTTATTAAAAACAAGCATTACAGGTCTTCTGCCGCCTCTAGATTGCCCTGCCCCAATTTCAAAAATAAAATCTTTTTCAAGCAGTGTATTTACTTGAGAGGAGACAGTAGACTTGTTTAATCCTGTAATCTCAGAGAGAGTTGCCCTGGAGACAGGGGAGTTCTTCAAAATTTCATCTAATATTAATTTTTGATTCATTTTTTTTACTAAAGCTTGATCTGCAATTTGAATAATAACCACTCCTTTGTTTATCCACCGAACTAAGTTGGTGTTTTTTGAAGCTTGAATTAGATATTTAAAAGTATCATATCTAATATTATAACTAAATTTTCTAAAAAAAACATTGAAATAAACATTTATTTTGTATATGATGAGATAAAGTTAGTTTATTGGATAAACAAACTAACTCAATTAAGATAGTTGATGGATAAACTTGTTCACTTAAATCAAAGGCTGAG -mRuby_with_RBS(Bsub): - description: mRuby with ribosomal binding site for Bacillus - prefix: AACT - suffix: ATCC - tags: [rbs_cds, reporter] - sequence: US_glmS: description: upstream glmS flank prefix: ATTA diff --git a/parts/parts/ecoli/ecoli.yaml b/parts/parts/ecoli/ecoli.yaml deleted file mode 100644 index a31b528..0000000 --- a/parts/parts/ecoli/ecoli.yaml +++ /dev/null @@ -1,103 +0,0 @@ -# CIDAR -eBFP2: - description: EBFP2 is a basic (constitutively fluorescent) blue fluorescent protein published in 2007, derived from Aequorea victoria. It is reported to be a rapidly-maturing monomer with moderate acid sensitivity. - prefix: - suffix: - references: ["https://www.fpbase.org/protein/ebfp2/"] - sequence: atggtgagcaagggcgaggagctgttcaccggggtggtgcccatcctggtcgagctggacggcgacgtaaacggccacaagttcagcgtgaggggcgagggcgagggcgatgccaccaacggcaagctgaccctgaagttcatctgcaccaccggcaagctgcccgtgccctggcccaccctcgtgaccaccctgagccacggcgtgcagtgcttcgcccgctaccccgaccacatgaagcagcacgacttcttcaagtccgccatgcccgaaggctacgtccaggagcgcaccatcttcttcaaggacgacggcacctacaagacccgcgccgaggtgaagttcgagggcgacaccctggtgaaccgcatcgagctgaagggcgtcgacttcaaggaggacggcaacatcctggggcacaagctggagtacaacttcaacagccacaacatctatatcatggccgtcaagcagaagaacggcatcaaggtgaacttcaagatccgccacaacgtggaggacggcagcgtgcagctcgccgaccactaccagcagaacacccccatcggcgacggccccgtgctgctgcccgacagccactacctgagcacccagtccgtgctgagcaaagaccccaacgagaagcgcgatcacatggtcctgctggagttccgcaccgccgccgggatcactctcggcatggacgagctgtacaagtaa -E1010m: - description: Fluorescent protein. Red. Modified from Bba_E1010 to fix illegal sites. - sequence: atggcttcctccgaggatgttatcaaagagttcatgcgtttcaaagttcgtatggaaggttccgttaacggtcacgagttcgaaatcgaaggtgaaggtgaaggtcgtccgtacgaaggtacccagaccgctaaactgaaagttaccaaaggtggtccgctgccgttcgcttgggacatcctgtccccgcagttccagtacggttccaaagcttacgttaaacacccggctgacatcccggactacctgaaactgtccttcccggaaggtttcaaatgggaacgtgttatgaacttcgaagatggtggtgttgttaccgttacccaggactcctccctgcaagacggtgagttcatctacaaagttaaactgcgtggtaccaacttcccgtccgacggtccggttatgcagaaaaaaaccatgggttgggaagcttccaccgaacgtatgtacccggaggatggtgctctgaaaggtgaaatcaaaatgcgtctgaaactgaaagacggtggtcactacgacgctgaagttaaaaccacctacatggctaaaaaaccggttcagctgccgggtgcttacaaaaccgacatcaaactggacatcacctcccacaacgaggactacaccatcgttgaacagtacgaacgtgctgaaggtcgtcactccaccggtgcttaataa -E0040m: - description: Fluorescent protein. Green. Modified from Bba_E0040 to fix illegal site. - sequence: atgcgtaaaggagaagaacttttcactggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggagagggtgaaggtgatgcaacatacggaaaacttacccttaaatttatttgcactactggaaaactacctgttccatggccaacacttgtcactactttcggttatggtgttcaatgctttgcgagatacccagatcatatgaaacagcatgactttttcaagagtgccatgcccgaaggttatgtacaggaaagaactatatttttcaaagatgacgggaactacaagacacgtgctgaagtcaagtttgaaggtgatacccttgttaatagaatcgagttaaaaggtattgattttaaagaagatggaaacattcttggacacaaattggaatacaactataactcacacaatgtatacatcatggcagacaaacaaaagaatggaatcaaagttaacttcaaaattagacacaacattgaagatggaagcgttcaactagcagaccattatcaacaaaatactccaattggcgatggccctgtccttttaccagacaaccattacctgtccacacaatctgccctttcgaaagatcccaacgaaaagagagatcacatggtccttcttgagtttgtaacagctgctgggattacacatggcatggatgaactatacaaataataa -E0030: - description: Fluorescent protein. Yellow. - sequence: atggtgagcaagggcgaggagctgttcaccggggtggtgcccatcctggtcgagctggacggcgacgtaaacggccacaagttcagcgtgtccggcgagggcgagggcgatgccacctacggcaagctgaccctgaagttcatctgcaccaccggcaagctgcccgtgccctggcccaccctcgtgaccaccttcggctacggcctgcaatgcttcgcccgctaccccgaccacatgaagctgcacgacttcttcaagtccgccatgcccgaaggctacgtccaggagcgcaccatcttcttcaaggacgacggcaactacaagacccgcgccgaggtgaagttcgagggcgacaccctggtgaaccgcatcgagctgaagggcatcgacttcaaggaggacggcaacatcctggggcacaagctggagtacaactacaacagccacaacgtctatatcatggccgacaagcagaagaacggcatcaaggtgaacttcaagatccgccacaacatcgaggacggcagcgtgcagctcgccgaccactaccagcagaacacccccatcggcgacggccccgtgctgctgcccgacaaccactacctgagctaccagtccgccctgagcaaagaccccaacgagaagcgcgatcacatggtcctgctggagttcgtgaccgccgccgggatcactctcggcatggacgagctgtacaagtaataa -cre: - description: cre/lox - sequence: atgtccaatttactgaccgtacaccaaaatttgcctgcattaccggtcgatgcaacgagtgatgaggttcgcaagaacctgatggacatgttcagggatcgccaggcgttttctgagcatacctggaaaatgcttctgtccgtttgccggtcgtgggcggcatggtgcaagttgaataaccggaaatggtttcccgcagaacctgaagatgttcgcgattatcttctatatcttcaggcgcgcggtctggcagtaaaaactatccagcaacatttgggccagctaaacatgcttcatcgtcggtccgggctgccacgaccaagtgacagcaatgctgtttcactggttatgcggcgaatccgaaaagaaaacgttgatgccggtgaacgtgcaaaacaggctctagcgttcgaacgcactgatttcgaccaggttcgttcactcatggaaaatagcgatcgctgccaggatatacgtaatctggcatttctggggattgcttataacaccctgttacgtatagccgaaattgccaggatcagggttaaagatatctcacgtactgacggtgggagaatgttaatccatattggcagaacgaaaacgctggttagcaccgcaggtgtagagaaggcacttagcctgggggtaactaaactggtcgagcgatggatttccgtctctggtgtagctgatgatccgaataactacctgttttgccgggtcagaaaaaatggtgttgccgcgccatctgccaccagccagctatcaactcgcgccctggaagggatttttgaagcaactcatcgattgatttacggcgctaaggatgactctggtcagagatacctggcctggtctggacacagtgcccgtgtcggagccgcgcgagatatggcccgcgctggagtttcaataccggagatcatgcaagctggtggctggaccaatgtaaatattgtcatgaactatatccgtaacctggatagtgaaacaggggcaatggtgcgcctgctggaagatggcgattag -araC: - description: Controller protein, araC repressor/activator (activates pBAD, I13453). Otherwise known as C0080 - sequence: atggctgaagcgcaaaatgatcccctgctgccgggatactcgtttaacgcccatctggtggcgggtttaacgccgattgaggccaatggttatctcgatttttttatcgaccgaccgctgggaatgaaaggttatattctcaatctcaccattcgcggtcagggggtggtgaaaaatcagggacgagaatttgtctgccgaccgggtgatattttgctgttcccgccaggagagattcatcactacggtcgtcatccggaggctcgcgaatggtatcaccagtgggtttactttcgtccgcgcgcctactggcatgaatggcttaactggccgtcaatatttgccaatacgggtttctttcgcccggatgaagcgcaccagccgcatttcagcgacctgtttgggcaaatcattaacgccgggcaaggggaagggcgctattcggagctgctggcgataaatctgcttgagcaattgttactgcggcgcatggaagcgattaacgagtcgctccatccaccgatggataatcgggtacgcgaggcttgtcagtacatcagcgatcacctggcagacagcaattttgatatcgccagcgtcgcacagcatgtttgcctgtcgccgtcgcgtctgtcacatcttttccgccagcagttagggattagcgtcttaagctggcgcgaggaccaacgcatcagccaggcgaagctgcttttgagcactacccggatgcctatcgccaccgtcggtcgcaatgttggttttgacgatcaactctatttctcgcgagtatttaaaaaatgcaccggggccagcccgagcgagttccgtgccggttgtgaagaaaaagtgaatgatgtagccgtcaagttgtcagctgcaaacgacgaaaactacgctttagtagcttaataa -luxR: - description: Controller protein, luxR repressor/activator (in concert with HSL, represses pLuxR(pR) R0063. Also up-regulates pLuxR(pL) R0062). Annotated as C0062. - sequence: atgaaaaacataaatgccgacgacacatacagaataattaataaaattaaagcttgtagaagcaataatgatattaatcaatgcttatctgatatgactaaaatggtacattgtgaatattatttactcgcgatcatttatcctcattctatggttaaatctgatatttcaatcctagataattaccctaaaaaatggaggcaatattatgatgacgctaatttaataaaatatgatcctatagtagattattctaactccaatcattcaccaattaattggaatatatttgaaaacaatgctgtaaataaaaaatctccaaatgtaattaaagaagcgaaaacatcaggtcttatcactgggtttagtttccctattcatacggctaacaatggcttcggaatgcttagttttgcacattcagaaaaagacaactatatagatagtttatttttacatgcgtgtatgaacataccattaattgttccttctctagttgataattatcgaaaaataaatatagcaaataataaatcaaacaacgatttaaccaaaagagaaaaagaatgtttagcgtgggcatgcgaaggaaaaagctcttgggatatttcaaaaatattaggttgcagtgagcgtactgtcactttccatttaaccaatgcgcaaatgaaactcaatacaacaaaccgctgccaaagtatttctaaagcaattttaacaggagcaattgattgcccatactttaaaaattaataa -tetR: - description: Controller protein, tetR repressor (represses pTet, C0040. can be inhibited by tetracyclin or aTc) - sequence: ATGtccagattagataaaagtaaagtgattaacagcgcattagagctgcttaatgaggtcggaatcgaaggtttaacaacccgtaaactcgcccagaagctaggtgtagagcagcctacattgtattggcatgtaaaaaataagcgggctttgctcgacgccttagccattgagatgttagataggcaccatactcacttttgccctttagaaggggaaagctggcaagattttttacgtaataacgctaaaagttttagatgtgctttactaagtcatcgcgatggagcaaaagtacatttaggtacacggcctacagaaaaacagtatgaaactctcgaaaatcaattagcctttttatgccaacaaggtttttcactagagaatgcattatatgcactcagcgctgtggggcattttactttaggttgcgtattggaagatcaagagcatcaagtcgctaaagaagaaagggaaacacctactactgatagtatgccgccattattacgacaagctatcgaattatttgatcaccaaggtgcagagccagccttcttattcggccttgaattgatcatatgcggattagaaaaacaacttaaatgtgaaagtgggtccgctgcaaacgacgaaaactacgctttagtagcttaataa -lacI: - description: Controller protein, lacI repressor (in concert with CAP, represses pLacI, R0010) - sequence: ATGatggtgaatgtgaaaccagtaacgttatacgatgtcgcagagtatgccggtgtctcttatcagaccgtttcccgcgtggtgaaccaggccagccacgtttctgcgaaaacgcgggaaaaagtggaagcggcgatggcggagctgaattacattcccaaccgcgtggcacaacaactggcgggcaaacagtcgttgctgattggcgttgccacctccagtctggccctgcacgcgccgtcgcaaattgtcgcggcgattaaatctcgcgccgatcaactgggtgccagcgtggtggtgtcgatggtagaacgaagcggcgtcgaagcctgtaaagcggcggtgcacaatcttctcgcgcaacgcgtcagtgggctgatcattaactatccgctggatgaccaggatgccattgctgtggaagctgcctgcactaatgttccggcgttatttcttgatgtctctgaccagacacccatcaacagtattattttctcccatgaggacggtacgcgactgggcgtggagcatctggtcgcattgggtcaccagcaaatcgcgctgttagcgggcccattaagttctgtctcggcgcgtctgcgtctggctggctggcataaatatctcactcgcaatcaaattcagccgatagcggaacgggaaggcgactggagtgccatgtccggttttcaacaaaccatgcaaatgctgaatgagggcatcgttcccactgcgatgctggttgccaacgatcagatggcgctgggcgcaatgcgcgccattaccgagtccgggctgcgcgttggtgcggatatctcggtagtgggatacgacgataccgaagatagctcatgttatatcccgccgttaaccaccatcaaacaggattttcgcctgctggggcaaaccagcgtggaccgcttgctgcaactctctcagggccaggcggtgaagggcaatcagctgttgcccgtctcactggtgaaaagaaaaaccaccctggcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcaggctgcaaacgacgaaaactacgctttagtagcttaataactctgatagtgctagtgtag -BCD12: - description: RBS - BiCistronic Design, medium strength (RBS part type, actually contains a small transcriptional unit and second RBS) - sequence: gggcccaagttcacttaaaaaggagatcaacaatgaaagcaattttcgtactgaaacatcttaatcatgctgcggagggtttct -B0032m: - description: RBS - Weiss RBS, medium strength. Modified from Bba_B0032 to adjust spacing in MC system - sequence: agagtcacacaggaaagtacta -BCD8: - description: RBS - BiCistronic Design, low strength (RBS part type, actually contains a small transcriptional unit and second RBS) - sequence: gggcccaagttcacttaaaaaggagatcaacaatgaaagcaattttcgtactgaaacatcttaatcatgcatcggaccgtttct -BCD2: - description: RBS - BiCistronic Design, high strength (RBS part type, actually contains a small transcriptional unit and second RBS) - sequence: gggcccaagttcacttaaaaaggagatcaacaatgaaagcaattttcgtactgaaacatcttaatcatgctaaggaggttttct -B0034m: - description: RBS - Weiss RBS, high strength. Modified from Bba_B0034 to adjust spacing in MC system. - sequence: agagaaagaggagaaatacta -B0033m: - description: RBS - Weiss RBS, low strength. Modified from Bba_B0033 to adjust spacing in MC system. - sequence: agagtcacacaggactacta -B0015: - description: Double terminator (B0010,B0012) - sequence: ccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttata -pBAD: - description: Controllable promoter - pBAD - araC regulated (pBAD promoter, inducible by expression of AraC, C0080) - seqeunce: acattgattatttgcacggcgtcacactttgctatgccatagcatttttatccataagattagcggatcctacctgacgctttttatcgcaactctctactgtttctccataccgtttttttgggctagc - -# -# Ecoflex -# - -eCFP(Ecoli): - description: placeholder - prefix: A - suffix: GGATCC - sequence: atggtgagcaagggcgaggagctgttcaccggggtggtgcccatcctggtcgagctggacggcgacgtgaacggccacaagttcagcgtgtccggcgagggcgagggcgatgccacctacggcaagctgaccctgaagttcatctgcaccaccggcaagctgcccgtgccctggcccaccctcgtgaccaccctgacctggggcgtgcagtgcttcagccgctaccccgaccacatgaagcagcacgacttcttcaagtccgccatgcccgaaggctacgtccaggagcgcaccatcttcttcaaggacgacggcaactacaagacccgcgccgaggtgaagttcgagggcgacaccctggtgaaccgcatcgagctgaagggcatcgacttcaaggaggacggcaacatcctggggcacaagctggagtacaactacatcagccacaacgtctatatcaccgccgacaagcagaagaacggcatcaaggccaacttcaagatccgccacaacatcgaggacggcagcgtgcagctcgccgaccactaccagcagaacacccccatcggcgacggccccgtgctgctgcccgacaaccactacctgagcacccagtccgccctgagcaaagaccccaacgagaagcgcgatcacatggtcctgctggagttcgtgaccgccgcc -eGFP(Ecoli): - description: placeholder - prefix: A - suffix: GGATCC - sequence: atgcgtaaaggagaagaacttttcactggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggagagggtgaaggtgatgcaacatacggaaaacttacccttaaatttatttgcactactggaaaactacctgttccatggccaacacttgtcactactttcggttatggtgttcaatgctttgcgagatacccagatcatatgaaacagcatgactttttcaagagtgccatgcccgaaggttatgtacaggaaagaactatatttttcaaagatgacgggaactacaagacacgtgctgaagtcaagtttgaaggtgatacccttgttaatagaatcgagttaaaaggtattgattttaaagaagatggaaacattcttggacacaaattggaatacaactataactcacacaatgtatacatcatggcagacaaacaaaagaatggaatcaaagttaacttcaaaattagacacaacattgaagatggaagcgttcaactagcagaccattatcaacaaaatactccaattggcgatggccctgtccttttaccagacaaccattacctgtccacacaatctgccctttcgaaagatcccaacgaaaagagagatcacatggtccttcttgagtttgtaacagctgctgggattacacatggcatggatgaactatacaaa -mCherry(Ecoli): - description: placeholder - prefix: A - suffix: GGATCC - sequence: atggtgagcaagggcgaggaggataacatggccatcatcaaggagttcatgcgcttcaaggtgcacatggagggctccgtgaacggccacgagttcgagatcgagggcgagggcgagggccgcccctacgagggcacccagaccgccaagctgaaggtgaccaagggtggccccctgcccttcgcctgggacatcctgtcccctcagttcatgtacggctccaaggcctacgtgaagcaccccgccgacatccccgactacttgaagctgtccttccccgagggcttcaagtgggagcgcgtgatgaacttcgaggacggcggcgtggtgaccgtgacccaggactcctccttgcaggacggcgagttcatctacaaggtgaagctgcgcggcaccaacttcccctccgacggccccgtaatgcagaagaagaccatgggctgggaggcctcctccgagcggatgtaccccgaggacggcgccctgaagggcgagatcaagcagaggctgaagctgaaggacggcggccactacgacgctgaggtcaagaccacctacaaggccaagaagcccgtgcagctgcccggcgcctacaacgtcaacatcaagttggacatcacctcccacaacgaggactacaccatcgtggaacagtacgaacgcgccgagggccgccactccaccggcggcatggacgagctgtacaag - -# -# CIDAR extension -# -J23101: - description: MoClo golden gate assembly AB part plasmid for J23101 (strength 1791x Anderson promoter; see http://parts.igem.org/Promoters/Catalog/Anderson) - sequence: TTTACAGCTAGCTCAGTCCTAGGTATTATGCTAGC -J23150: - description: MoClo golden gate assembly AB part for J23150 (strength ~400x Anderson promoter; see http://parts.igem.org/Promoters/Catalog/Anderson) - sequence: TGCTGTTCCGCTGGGCATGCTTTACGGCTAGCTCAGTCCTAGGTATTATGC -J23151: - description: MoClo golden gate assembly AB part for J23151 (strength ~900x Anderson promoter; see http://parts.igem.org/Promoters/Catalog/Anderson) - sequence: TGCTGTTCCGCTGGGCATGCTTGATGGCTAGCTCAGTCCTAGGTACAATGC -pOR1-OR2: - description: MoClo golden gate assembly AB part for pOR1-OR2 (lambda cI-repressable strong promoter) - sequence: TGCTGTTCCGCTGGGCATGCTGAGCTAACACCGTGCGTGTTGACAATTTTACCTCTGGCGGTGATAATGGTTGCAGC -Ptet: - description: MoClo golden gate assembly AB part for pLas-TetO (Promoter activated by LasR and repressed by TetR; encodes <3OC12 AHL AND aTc> logic). - sequence: CTAGCAAGGGTCCGGGTTCACCGAAATCTATCTCATTTGCTAGTTATAAAATTATGAAATTTGCGTAAATTCCCTATCAGTGATAGAGATTCAGAAGCAAT -pCin-TetO: - description: MoClo golden gate assembly AB part for pCin-TetO (Promoter activated by CinR and repressed by TetR; encodes <3OC14 AHL AND aTc> logic). - sequence: CGGAACTCTTGTGCGTGGCGCATGTCGTTATCGCAAAACCGCTGCACACTTTTGCGCGACATGCTCTGATCCCCCTCATCTGGGGGGGCCTATCTGAGGGAATTTACTTCCCTATCAGTGATAGAGATAATGAGCACATCCCTATCAGTGATAGAGATG -pCin-LacO: - description: MoClo golden gate assembly AB part for pCin-LacO (Promoter activated by CinR and repressed by LacI; encodes <3OC14 AHL AND IPTG> logic). - sequence: CGGAACTCTTGTGCGTGGCGCATGTCGTTATCGCAAAACCGCTGCACACTTTTGCGCGACATGCTCTGATCCCCCTCATCTGGGGGGGCCTATCTGAGGGAATTTACTTGTGAGCGCTCACAATTGATAATGAGCACAATTGTGAGCGGATAACAATTGCT -T7_promoter: - description: The T7 promoter binding site - sequence: TAATACGACTCACTATAG diff --git a/parts/parts/ecoli/origins.yaml b/parts/parts/ecoli/origins.yaml deleted file mode 100644 index 17a3808..0000000 --- a/parts/parts/ecoli/origins.yaml +++ /dev/null @@ -1,52 +0,0 @@ -# Origins -ColE1: - description: High copy (500-700 copies) common Escherichia coli origin - sequence: TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA -RSF1030: - description: High copy (>100 copies) Escherichia coli origin. Compatible with pUC/pBR322, p15A, ColA, CloDF13, and SC101 - sequence: AACGGAATAGCTGTTCGTTGACTTGATAGACCGATTGATTCATCATCTCATAAATAAAGAAAAACCACCGCTACCAACGGTGGTTTTCTCAAGGTTCGCTGAGCTACCAACTCTTTGAACCAAGGTAAGTGGGTTGGAGGACCGCACTCACCAAAATCTGTTCTTTCAGTTTAGCCTTAACAGGTGCATAACTTCAAGACAAAGTCCTCTAAATCAGTTACCAATGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCAACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGGCTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCTGCGGCGTTGGCTTCTTCCGGTGCTTTGCTTTTTGCTCACATGTTCTTTCCGGCTTTATCCCCTGATTCTGTGGATAACCGTATTACCGCTTTTGAGTGAGCTGACACCGCTCGCCGCAGTCGAACGACCGAGCGTAGCGAGTCAGTGAGCGAGGAAGCGGAAGA -pBR322: - description: Medium copy (~20) common Escherichia coli origin - sequence: CCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGTGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGTATACACTCCGCTATCGCTACGTGACTGGGTCATGGCTGCGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGGCAGCTGCGGTAAAGCTCATCAGCGTGGTCGTGCAGCGATTCACAGATGTCTGCCTGTTCATCCGCGTCCAGCTCGTTGAGTTTCTCCAGAAGCGTTAATGTCTGGCTTCTGATAAAGCGGGCCATGTTAAGGGCGGTTTTTTCCTGTTTGGTCACTGATGCCTCCGTGTAAGGGGGATTTCTGTTCATGGGGGTAATGATACCGATGAAACGAGAGAGGATGCTCACGATACGGGTTACTGATGATGAACATGCCCGGTTACTGGAACGTTGTGAGGGTAAACAACTGGCGGTATGGATGCGGCGGG -p15A: - description: Medium copy (10-12 copies) common Escherichia coli origin. pUC/pBR322, SC101, ColA, CloDF13, and RSF1030 - sequence: TTGAGATCGTTTTGGTCTGCGCGTAATCTCTTGCTCTGAAAACGAAAAAACCGCCTTGCAGGGCGGTTTTTCGAAGGTTCTCTGAGCTACCAACTCTTTGAACCGAGGTAACTGGCTTGGAGGAGCGCAGTCACCAAAACTTGTCCTTTCAGTTTAGCCTTAACCGGCGCATGACTTCAAGACTAACTCCTCTAAATCAATTACCAGTGGCTGCTGCCAGTGGTGCTTTTGCATGTCTTTCCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGACTGAACGGGGGGTTCGTGCATACAGTCCAGCTTGGAGCGAACTGCCTACCCGGAACTGAGTGTCAGGCGTGGAATGAGACAAACGCGGCCATAACAGCGGAATGACACCGGTAAACCGAAAGGCAGGAACAGGAGAGCGCACGAGGGAGCCGCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCACTGATTTGAGCGTCAGATTTCGTGATGCTTGTCAGGGGGGCGGAGCCTATGGAAA -ColE2: - description: Medium copy plasmid. Can be engineered easily for different copy number levels. Orthogonal to R6K origin - sequence: TTAAGGGGGGCGTACAACGGGAGTTATGGTAAATGGATCGGGTTTTCGGGAAGGATCGACAGGATTTGCCGTTGGGTGTAGTGTAAGCGACTGAAAAACAAACGCCCCGTAAATCGTGCTCTCACCGCCAAGATTGATCACGAAATTACAGGGCGCCGGGTTCCGCGTTTCCCGATGGGAAAGCGCGGTTAGTTAAACTGTGTACCGAGAGAAATCGTATCACATGAGCGCCGTACTTCAACGCTTCAGGGAAAAATTACCGCACAAACCGTACTGTACGAACGATTTCGCGTACGGCGTTCGCATTCTGCCGAAAAACATTGCCATTCTTGCCCGTTTCATCCAGCAGAACCAGCCACATGCACTGTACTGGCTTCCCTTTGACGTGGACCGGACGGGGGCATCAATCGACTGGAGCGACCGGAATTGTCCGGCCCCGAACATCACCGTAAAAAATCCCCGTAACGGGCACGCGCATCTGCTCTACGCGCTCGCCCTTCCTGTGAGAACTGCGCCGGATGCATCGGCTTCGGCGCTCAGATACGCTGCCGCTATTGAGCGTGCGTTGTGTGAAAAACTGGGCGCGGATGTGAATTACAGCGGCCTGATCTGCAAAAATCCGTGCCACCCTGAATGGCAGGAAGTGGAATGGCGCGAGGAACCCTACACTCTCGACGAACTGGCTGATTATCTCGATTTGAGCGCCTCAGCGCGCCGTAGCGTCGATAAAAATTACGGGCTGGGGCGAAACTACCATCTGTTCGAAAAGGTCCGTAAATGGGCCTACAGAGCGATTCGTCAGGGCTGGCCTGTATTCTCACAATGGCTTGATGCCGTGATCCAGCGTGTCGAAATGTACAACGCATCGCTTCCCGTTCCGCTTTCTCCGGCTGAATGTCGGGCTATTGGCAAGAGCATTGCGAAATATACACACAGGAAATTCTCACCAGAGGGATTTTCCGCTGTACAGGCCGCTCGAGGTCGCAAGGGCGGAACTAAATCTAAGCGCGCAGCAGTTCCTACATCAGCACGTTCGCTGAAGCCGTGGGAGGCATTAGGGATCAGTCGAGCGACGTACTACCGAAAATTAAAATGTGATCCAGATCTCGCAAAATAAGACCAGATAAGCCTTATCAGATAACAGCGCCCTTTTGGCGTCTTTTTGAGCAGTGATCTTTCTTGCGATTTTCTCCGTTCTGC -pWV01: - description: Medium copy broad host range plasmid. - sequence: CGATTTTTTATTAAAATTTTAGCGTTTATTTCGTTTAGTTATCGGCATAATCGTTAAAACAGGCGTTATCGTAGCGTAAAAGCCCTTGAGCGTAGCGTGGCTTTGCAGCGAAGATGTTGTCTGTTAGATTATGAAAGCCGATGACTGAATGAAATAATAAGCGCAGCGCCCTTCTATTTCGGTTGGAGGAGGCTCAAGGGAGTATGAGGGAATGAAATTCCCTCATGGGTTTGATTTTAAAAATTGCTTGCAATTTTGCCGAGCGGTAGCGCTGGAAAATTTTTGAAAAAAATTTGGAATTTGGAAAAAAATGGGGGGAAAGGAAGCGAATTTTGCTTCCGTACTACGACCCCCCATTAAGTGCCGAGTGCCAATTTTTGTGCCAAAAACGCTCTATCCCAACTGGCTCAAGGGTTTAAGGGGTTTTTCAATCGCCAACGAATCGCCAACGTTTTCGCCAACGTTTTTTATAAATCTATATTTAAGTAGCTTTATTGTTGTTTTTATGATTACAAAGTGATACACTAACTTTATAAAATTATTTGATTGGAGTTTTTTAAATGGTGATTTCAGAATCGAAAAAAAGAGTTATGATTTCTCTGACAAAAGAGCAAGATAAAAAATTAACAGATATGGCGAAACAAAAAGGTTTTTCAAAATCTGCGGTTGCGGCGTTAGCTATAGAAGAATATGCAAGAAAGGAATCAGAACAAAAAAAATAAGCGAAAGCTCGCGTTTTTAGAAGGATACGAGTTTTCGCTACTTGTTTTTGATAAGGTAATTATATCATGGCTATTAAAAATACTAAAGCTAGAAATTTTGGATTTTTATTATATCCTGACTCAATTCCTAATGATTGGAAAGAAAAATTAGAGAGTTTGGGCGTATCTATGGCTGTCAGTCCTTTACACGATATGGACGAAAAAAAAGATAAAGATACATGGAATAGTAGTGATGTTATACGAAATGGAAAGCACTATAAAAAACCACACTATCACGTTATATATATTGCACGAAATCCTGTAACAATAGAAAGCGTTAGGAACAAGATTAAGCGAAAATTGGGGAATAGTTCAGTTGCTCATGTTGAGATACTTGATTATATCAAAGGTTCATATGAATATTTGACTCATGAATCAAAGGACGCTATTGCTAAGAATAAACATATATACGACAAAAAAGATATTTTGAACATTAATGATTTTGATATTGACCGCTATATAACACTTGATGAAAGCCAAAAAAGAGAATTGAAGAATTTACTTTTAGATATAGTGGATGACTATAATTTGGTAAATACAAAAGATTTAATGGCTTTTATTCGCCTTAGGGGAGCGGAGTTTGGAATTTTAAATACGAATGATGTAAAAGATATTGTTTCAACAAACTCTAGCGCCTTTAGATTATGGTTTGAGGGCAATTATCAGTGTGGATATAGAGCAAGTTATGCAAAGGTTCTTGATGCTGAAACGGGGGAAATAAAATGACAAACAAAGAAAAAGAGTTATTTGCTGAAAATGAGGAATTAAAAAAAGAAATTAAGGACTTAAAAGAGCGTATTGAAAGATACAGAGAAATGGAAGTTGAATTAAGTACAACAATAGATTTATTGAGAGGAGGGATTATTGAATAAATAAAAGCCCCCTGACGAAAGTCGAAGGGGGTTTTTATTTTGGTTTGATGTTGCGATTAATAGCAATACAATTGCAATAAACAAAATGATC -pBAV1KT5: - description: High copy (~350 E.coli, ~70 Bacillus subtilis, ~3 Agrobacterium tumefaciens, ~60 Acinetobacter baylyi, 100-500 Streptococcus pneumoniae) broad host range vector derived from pWV01. - description: CCAAGCACTAGGCGATTTTTTATTAAAATTTTAGCGTTTATTTCGTTTAGTTATCGGCATAATCGTTAAAACAGGCGTTATCGTAGCGTAAAAGCCCTTGAGCGTAGCGTGGCTTTGCAGCGAAGATGTTGTCTGTTAGATTATGAAAGCCGATGACTGAATGAAATAATAAGCGCAGCGCCCTTCTATTTCGGTTGGAGGAGGCTCAAGGGAGTATGAGGGAATGAAATTCCCTCATGGGTTTGATTTTAAAAATTGCTTGCAATTTTGCCGAGCGGTAGCGCTGGAAAATTTTTGAAAAAAATCTGGAATTTGGAAAAAAATGGGGGGAAAGGAAGCGAATTTTGCTTCCGTACTACGACCCCCCATTAAGTGCCGAGTGCCAATTTTTGTGCCAAAAACGCTCTATCCCAACTGGCTCAAGGGTTTAAGGGGTTTTTCAATCGCCAACGAATCGCCAACGTTTTCGCCAACGTTTTTTATAAATCTATATTTAAGTAGCTTTATTGTTGTTTTTATGATTACAAAGTGATACACTAACTTTATAAAATTATTTGATTGGAGTTTTTTAAATGGTGATTTCAGAATCGAAAAAAAGAGTTATGATTTCTCTGACAAAAGAGCAAGATAAAAAATTAACAGATATGGCGAAACAAAAAGGTTTTTCAAAATCTGCGGTTGCGGCGTTAGCTATAGAAGAATATGCAAGAAAGGAATCAGAACAAAAAAAATAAGCGAAAGCTCGCGTTTTTAGAAGGATACGAGTTTTCGCTACTTGTTTTTGATAAGGTAATTATATCATGGCTATTAAAAATACTAAAGCTAGAAATTTTGGATTTTTATTATATCCTGACTCAATTCCTAATGATTGGAAGGAAAAATTAGAGAGTTTGGGCGTATCTATGGCTGTCAGTCCTTTACACGATATGGACGAAAAAAAAGATAAAGATACATGGAATAGTAGTGATGTTATACGAAATGGAAAGCACTATAAAAAACCACACTATCACGTTATATATATTGCACGAAATCCTGTAACAATAGAAAGCGTTAGGAACAAGATTAAGCGAAAATTGGGGAATAGTTCAGTTGCTCATGTTGAGATACTTGATTATATCAAAGGTTCATATGAATATTTGACTCATGAATCAAAGGACGCTATTGCTAAGAATAAACATATATACGACAAAAAAGATATTTTGAACATTAATGATTTTGATATTGACCGCTATATAACACTTGATGAAAGCCAAAAAAGAGAATTGAAGAATTTACTCTTAGATATAGTGGATGACTATAATTTGGTAAATACAAAAGATTTAATGGCTTTTATTCGCCTTAGGGGAGCGGAGTTTGGAATTTTAAATACGAATGATGTAAAAGATATTGTTTCAACAAACTCTAGCGCCTTTAGATTATGGTTTGAGGGCAATTATCAGTGTGGATATAGAGCAAGTTATGCAAAGGTTCTTGATGCTGAAACGGGGGAAATAAAATGACACTAGGTCTAGGGCGGCGGATTTGTCCTACTCAGGAGAGCGTTCACCGACAAACAACAGATAAAACGAAAGGCCCAGTCTTTCGACTGAGCCTTTCGTTTTATTTG -RSF1010: - description: Low-med copy very broad host range origin (most gram-negative and some gram-positive) - sequence: GTCCTGGCCGCCACATGAGGCATGTTCCGCGTCCTTGCAATACTGTGTTTACATACAGTCTATCGCTTAGCGGAAAGTTCTTTTACCCTCAGCCGAAATGCCTGCCGTTGCTAGACATTGCCAGCCAGTGCCCGTCACTCCCGTACTAACTGTCACGAACCCCTGCAATAACTGTCACGCCCCCCTGCAATAACTGTCACGAACCCCTGCAATAACTGTCACGCCCCCAAACCTGCAAACCCAGCAGGGGCGGGGGCTGGCGGGGTGTTGGAAAAATCCATCCATGATTATCTAAGAATAATCCACTAGGCGCGGTTATCAGCGCCCTTGTGGGGCGCTGCTGCCCTTGCCCAATATGCCCGGCCAGAGGCCGGATAGCTGGTCTATTCGCTGCGCTAGGCTACACACCGCCCCACCGCTGCGCGGCAGGGGGAAAGGCGGGCAAAGCCCGCTAAACCCCACACCAAACCCCGCAGAAATACGCTGGAGCGCTTTTAGCCGCTTTAGCGGCCTTTCCCCCTACCCGAAGGGTGGGGGCGCGTGTGCAGCCCCGCAGGGCCTGTCTCGGTCGATCATTCAGCCCGGCTCATCCTTCTGGCGTGGCGGCAGACCGAACAAGGCGCGGTCGTGGTCGCGTTCAAGGTACGCATCCATTGCCGCCATGAGCCGATCCTCCGGCCACTCGCTGCTGTTCACCTTGGCCAAAATCATGGCCCCCACCAGCACCTTGCGCCTTGTTTCGTTCTTGCGCTCTTGCTGCTGTTCCCTTGCCCGCACCCGCTGAATTTCGGCATTGATTCGCGCTCGTTGTTCTTCGAGCTTGGCCAGCCGATCCGCCGCCTTGTTGCTCCCCTTAACCATCTTGACACCCCATTGTTAATGTGCTGTCTCGTAGGCTATCATGGAGGCACAGCGGCGGCAATCCCGACCCTACTTTGTAGGGGAGGGCGCACTTACCGGTTTCTCTTCGAGAAACTGGCCTAACGGCCACCCTTCGGGCGGTGCGCTCTCCGAGGGCCATTGCATGGAGCCGAAAAGCAAAAGCAACAGCGAGGCAGCATGGCGATTTATCACCTTACGGCGAAAACCGGCAGCAGGTCGGGCGGCCAATCGGCCAGGGCCAAGGCCGACTACATCCAGCGCGAAGGCAAGTATGCCCGCGACATGGATGAAGTCTTGCACGCCGAATCCGGGCACATGCCGGAGTTCGTCGAGCGGCCCGCCGACTACTGGGATGCTGCCGACCTGTATGAACGCGCCAATGGGCGGCTGTTCAAGGAGGTCGAATTTGCCCTGCCGGTCGAGCTGACCCTCGACCAGCAGAAGGCGCTGGCGTCCGAGTTCGCCCAGCACCTGACCGGTGCCGAGCGCCTGCCGTATACGCTGGCCATCCATGCCGGTGGCGGCGAGAACCCGCACTGCCACCTGATGATCTCCGAGCGGATCAATGACGGCATCGAGCGGCCCGCCGCTCAGTGGTTCAAGCGGTACAACGGCAAGACCCCGGAGAAGGGCGGGGCACAAAAGACCGAAGCGCTCAAGCCCAAGGCATGGCTTGAGCAGACCCGCGAGGCATGGGCCGACCATGCCAACCGGGCATTAGAGCGGGCTGGCCACGACGCCCGCATTGACCACAGAACACTTGAGGCGCAGGGCATCGAGCGCCTGCCCGGTGTTCACCTGGGGCCGAACGTGGTGGAGATGGAAGGCCGGGGCATCCGCACCGACCGGGCAGACGTGGCCCTGAACATCGACACCGCCAACGCCCAGATCATCGACTTACAGGAATACCGGGAGGCAATAGACCATGAACGCAATCGACAGAGTGAAGAAATCCAGAGGCATCAACGAGTTAGCGGAGCAGATCGAACCGCTGGCCCAGAGCATGGCGACACTGGCCGACGAAGCCCGGCAGGTCATGAGCCAGACCCAGCAGGCCAGCGAGGCGCAGGCGGCGGAGTGGCTGAAAGCCCAGCGCCAGACAGGGGCGGCATGGGTGGAGCTGGCCAAAGAGTTGCGGGAGGTAGCCGCCGAGGTGAGCAGCGCCGCGCAGAGCGCCCGGAGCGCGTCGCGGGGGTGGCACTGGAAGCTATGGCTAACCGTGATGCTGGCTTCCATGATGCCTACGGTGGTGCTGCTGATCGCATCGTTGCTCTTGCTCGACCTGACGCCACTGACAACCGAGGACGGCTCGATCTGGCTGCGCTTGGTGGCCCGATGAAGAACGACAGGACTTTGCAGGCCATAGGCCGACAGCTCAAGGCCATGGGCTGTGAGCGCTTCGATATCGGCGTCAGGGACGCCACCACCGGCCAGATGATGAACCGGGAATGGTCAGCCGCCGAAGTGCTCCAGAACACGCCATGGCTCAAGCGGATGAATGCCCAGGGCAATGACGTGTATATCAGGCCCGCCGAGCAGGAGCGGCATGGTCTGGTGCTGGTGGACGACCTCAGCGAGTTTGACCTGGATGACATGAAAGCCGAGGGCCGGGAGCCTGCCCTGGTAGTGGAAACCAGCCCGAAGAACTATCAGGCATGGGTCAAGGTGGCCGACGCCGCAGGCGGTGAACTTCGGGGGCAGATTGCCCGGACGCTGGCCAGCGAGTACGACGCCGACCCGGCCAGCGCCGACAGCCGCCACTATGGCCGCTTGGCGGGCTTCACCAACCGCAAGGACAAGCACACCACCCGCGCCGGTTATCAGCCGTGGGTGCTGCTGCGTGAATCCAAGGGCAAGACCGCCACCGCTGGCCCGGCGCTGGTGCAGCAGGCTGGCCAGCAGATCGAGCAGGCCCAGCGGCAGCAGGAGAAGGCCCGCAGGCTGGCCAGCCTCGAACTGCCCGAGCGGCAGCTTAGCCGCCACCGGCGCACGGCGCTGGACGAGTACCGCAGCGAGATGGCCGGGCTGGTCAAGCGCTTCGGTGATGACCTCAGCAAGTGCGACTTTATCGCCGCGCAGAAGCTGGCCAGCCGGGGCCGCAGTGCCGAGGAAATCGGCAAGGCCATGGCCGAGGCCAGCCCAGCGCTGGCAGAGCGCAAGCCCGGCCACGAAGCGGATTACATCGAGCGCACCGTCAGCAAGGTCATGGGTCTGCCCAGCGTCCAGCTTGCGCGGGCCGAGCTGGCACGGGCACCGGCACCCCGCCAGCGAGGCATGGACAGGGGCGGGCCAGATTTCAGCATGTAGTGCTTGCGTTGGTACTCACGCCTGTTATACTATGAGTACTCACGCACAGAAGGGGGTTTTATGGAATACGAAAAAAGCGCTTCAGGGTCGGTCTACCTGATCAAAAGTGACAAGGGCTATTGGTTGCCCGGTGGCTTTGGTTATACGTCAAACAAGGCCGAGGCTGGCCGCTTTTCAGTCGCTGATATGGCCAGCCTTAACCTTGACGGCTGCACCTTGTCCTTGTTCCGCGAGGACAAGCCTTTCGGCCCCGGCAAGTTTCTCGGTGACTGATATGAAAGACCAAAAGGACAAGCAGACCGGCGACCTGCTGGCCAGCCCTGACGCTGTACGCCAAGCGCGATATGCCGAGCGCATGAAGGCCAAAGGGATGCGTCAGCGCAAGTTCTGGCTGACCGACGACGAATACGAGGCGCTGCGCGAGTGCCTGGAAGAACTCAGAGCGGCGCAGGGCGGGGGTAGTGACCCCGCCAGCGCCTAACCACCAACTGCCTGCAAAGGAGGCAATCAATGGCTACCCATAAGCCTATCAATATTCTGGAGGCGTTCGCAGCAGCGCCGCCACCGCTGGACTACGTTTTGCCCAACATGGTGGCCGGTACGGTCGGGGCGCTGGTGTCGCCCGGTGGTGCCGGTAAATCCATGCTGGCCCTGCAACTGGCCGCACAGATTGCAGGCGGGCCGGATCTGCTGGAGGTGGGCGAACTGCCCACCGGCCCGGTGATCTACCTGCCCGCCGAGGACCCGCCCACCGCCATTCATCACCGCCTGCACGCCCTTGGGGCGCACCTCAGCGCCGAGGAACGGCAAGCCGTGGCTGACGGCCTGCTGATCCAGCCGCTGATCGGCAGCCTGCCCAACATCATGGCCCCGGAGTGGTTCGACGGCCTCAAGCGCGCCGCCGAGGGCCGCCGCCTGATGGTGCTGGACACGCTGCGCCGGTTCCACATCGAGGAAGAAAACGCCAGCGGCCCCATGGCCCAGGTCATCGGTCGCATGGAGGCGATCGCCGCCGATACCGGGTGCTCTATCGTGTTCCTGCACCATGCCAGCAAGGGCGCGGCCATGATGGGCGCAGGCGACCAGCAGCAGGCCAGCCGGGGCAGCTCGGTACTGGTCGATAACATCCGCTGGCAGTCCTACCTGTCGAGCATGACCAGCGCCGAGGCCGAGGAATGGGGTGTGGACGACGACCAGCGCCGGTTCTTCGTCCGCTTCGGTGTGAGCAAGGCCAACTATGGCGCACCGTTCGCTGATCGGTGGTTCAGGCGGCATGACGGCGGGGTGCTCAAGCCCGCCGTGCTGGAGAGGCAGCGCAAGAGCAAGGGGGTGCCCCGTGGTGAAGCCTAAGAACAAGCACAGCCTCAGCCACGTCCGGCACGACCCGGCGCACTGTCTGGCCCCCGGCCTGTTCCGTGCCCTCAAGCGGGGCGAGCGCAAGCGCAGCAAGCTGGACGTGACGTATGACTACGGCGACGGCAAGCGGATCGAGTTCAGCGGCCCGGAGCCGCTGGGCGCTGATGATCTGCGCATCCTGCAAGGGCTGGTGGCCATGGCTGGGCCTAATGGCCTAGTGCTTGGCCCGGAACCCAAGACCGAAGGCGGACGGCAGCTCCGGCTGTTCCTGGAACCCAAGTGGGAGGCCGTCACCGCTGAATGCCATGTGGTCAAAGGTAGCTATCGGGCGCTGGCAAAGGAAATCGGGGCAGAGGTCGATAGTGGTGGGGCGCTCAAGCACATACAGGACTGCATCGAGCGCCTTTGGAAGGTATCCATTATCGCCCAGAATGGCCGCAAGCGGCAGGGGTTTCGGCTGCTGTCGGAGTACGCCAGCGACGAGGCGGACGGGCGCCTGTACGTGGCCCTGAACCCCTTGATCGCGCAGGCCGTCATGGGTGGCGGCCAGCATGTGCGCATCAGCATGGACGAGGTGCGGGCGCTGGACAGCGAAACCGCCCGCCTGCTGCACCAGCGGCTGTGTGGCTGGATCGACCCCGGCAAAACCGGCAAGGCTTCCATAGATACCTTGTGCGGCTATGTCTGGCCGTCAGAGGCCAGTGGTTCGACCATGCGCAAGCGCCGCCAGCGGGTGCGCGAGGCGTTGCCGGAGCTGGTCGCGCTGGGCTGGACGGTAACCGAGTTCGCGGCGGGCAAGTACGACATCACCCGGCCCAAGGCGGCAGGCTGACCCCCCCCACTCTATTGTAAACAAGACATTTTTATCTTTTATATTCAATGGCTTATTTTCCTGCTAATTGGTAATACCATGAAAAATACCATGCTCAGAAAAGGCTTAACAATATTTTGAAAAATTGCCTACTGAGCGCTGCCGCACAGCTCCATAGGCCGCTTTCCTGGCTTTGCTTCCAGATGTATGCTCTTATGCTCC -SC101: - description: Low copy (~5 copies) Escherichia coli origin. Compatible with pUC/pBR322, p15A, ColA, CloDF13, and RSF1030 - sequence: ATGCAGCGATTTGCCCGAGCTTGCGAGGGTGCTACTTAAGCCTTTAGGGTTTTAAGGTCTGTTTTGTAGAGGAGCAAACAGCGTTTGCGACATCCTTTTGTAATACTGCGGAACTGACTAAAGTAGTGAGTTATACACAGGGCTGGGATCTATTCTTTTTATCTTTTTTTATTCTTTCTTTATTCTATAAATTATAACCACTTGAATATAAACAAAAAAAACACACAAAGGTCTAGCGGAATTTACAGAGGGTCTAGCAGAATTTACAAGTTTTCCAGCAAAGGTCTAGCAGAATTTACAGATACCCACAACTCAAAGGAAAAGGACTAGTAATTATCATTGACTAGCCCATCTCAATTGGTATAGTGATTAAAATCACCTAGACCAATTGAGATGTATGTCTGAATTAGTTGTTTTCAAAGCAAATGAACTAGCGACTAGTCGCTATGACTTAACGGAGCATGAAACCAAGCTAATTTTATGCTGTGTGGCACTACTCAACCCCACGATTGAAAACCCTACAAGGAAAGAACGGACGGTATCGTTCACTTATAACCAATACGTTCAGATGATGAACATCAGTAGGGAAAATGCTTATGGTGTATTAGCTAAAGCAACCAGAGAGCTGATGACGAGAACTGTGGAAATCAGGAATCCTTTGGTTAAAGGCTTTGAGATTTTCCAGTGGACAAACTATGCCAAGTTCTCAAGCGAAAAATTAGAATTAGTTTTTAGTGAAGAGATATTGCCTTATCTTTTCCAGTTAAAAAAATTCATAAAATATAATCTGGAACATGTTAAGTCTTTTGAAAACAAATACTCTATGAGGATTTATGAGTGGTTATTAAAAGAACTAACACAAAAGAAAACTCACAAGGCAAATATAGAGATTAGCCTTGATGAATTTAAGTTCATGTTAATGCTTGAAAATAACTACCATGAGTTTAAAAGGCTTAACCAATGGGTTTTGAAACCAATAAGTAAAGATTTAAACACTTACAGCAATATGAAATTGGTGGTTGATAAGCGAGGCCGCCCGACTGATACGTTGATTTTCCAAGTTGAACTAGATAGACAAATGGATCTCGTAACCGAACTTGAGAACAACCAGATAAAAATGAATGGTGACAAAATACCAACAACCATTACATCAGATTCCTACCTACATAACGGACTAAGAAAAACACTACACGATGCTTTAACTGCAAAAATTCAGCTCACCAGTTTTGAGGCAAAATTTTTGAGTGACATGCAAAGTAAGTATGATCTCAATGGTTCGTTCTCATGGCTCACGCAAAAACAACGAACCACACTAGAGAACATACTGGCTAAATACGGAAGGATCTGAGGTTCTTATGGCTCTTGTATCTATCAGTGAAGCATCAAGACTAACAAACAAAAGTAGAACAACTGTTCACCGTTACATATCAAAGGGAAAACTGTCCATATGCACAGATGAAAACGGTGTAAAAAAGATAGATACATCAGAGCTTTTACGAGTTTTTGGTGCA -pRK2: - description: Low copy (4-7 copies) origin with broad gram-negative host range - sequence: GTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGGGGGATCAGGACCGCTGCCGGAGCGCAACCCACTCACTACAGCAGAGCCATGTAGGGCCGCCGGCGTTGTGGATACCTCGCGGAAAACTTGGCCCTCACTGACAGATGAGGGGCGGACGTTGACACTTGAGGGGCCGACTCACCCGGCGCGGCGTTGACAGATGAGGGGCAGGCTCGATTTCGGCCGGCGACGTGGAGCTGGCCAGCCTCGCAAATCGGCGAAAACGCCTGATTTTACGCGAGTTTCCCACAGATGATGTGGACAAGCCTGGGGATAAGTGCCCTGCGGTATTGACACTTGAGGGGCGCGACTACTGACAGATGAGGGGCGCGATCCTTGACACTTGAGGGGCAGAGTGCTGACAGATGAGGGGCGCACCTATTGACATTTGAGGGGCTGTCCACAGGCAGAAAATCCAGCATTTGCAAGGGTTTCCGCCCGTTTTTCGGCCACCGCTAACCTGTCTTTTAACCTGCTTTTAAACCAATATTTATAAACCTTGTTTTTAACCAGGGCTGCGCCCTGTGCGCGTGACCGCGCACGCCGAAGGGGGGTGCCCCCCCTTCTCGAACCCTCCCGGCCCGCTAACGCGGGCCTCCCATCCCCCCAGGGGCTGCGCCCCTCGGCCGCGAACGGCCTCACCCCAAAAATGGCAGCCACGTAGAAAGCCAGTCCGCAGAAACGGTGCTGACCCCGGATGAATGTCAGCTACTGGGCTATCTGGACAAGGGAAAACGCAAGCGCAAAGAGAAAGCAGGTAGCTTGCAGTGGGCTTACATGGCGATAGCTAGACTGGGCGGTTTTATGGACAGCAAGCGAACCGGAATTGCCAGCTGGGGCGCCCTCTGGTAAGGTTGGGAAGCCCTGCAAAGTAAACTGGATGGCTTTCTTGCCGCCAAGGATCTGATGGCGCAGGGGATCAAGATCGACGGATCGATCCGGGGAATTAATTCCGGGGCAATCCCGCAAGGAGGGTGAATGAATCGGACGTTTGACCGGAAGGCATACAGGCAAGAACTGATCGACGCGGGGTTTTCCGCCGAGGATGCCGAAACCATTGCAAGCCGCACCGTCATGCGTGCGCCCCGCGAAACCTTCCAGTCCGTCGGCTCGATAGTCCAGCAAGCTACGGCCAAGATCGAGCGCGACAGCGTGCAACTGGCTCCCCCTGCCCTGCCCGCGCCATCGGCCGCCGTGGAGCGTTCGCGTCGCCTCGAACAGGAGGCGGCAGGTTTGGCGAAGTCGATGACCATCGACACGCGAGGAACTATGACGACCAAGAAGCGAAAAACCGCCGGCGAGGACCTGGCAAAACAGGTCAGCGAAGCCAAGCAGGCCGCGTTGCTGAAACACACGAAGCAGCAGATCAAGGAAATGCAGCTTTCCTTGTTCGATATTGCGCCGTGGCCGGACACGATGCGAGCCATGCCAAACGACACGGCCCGCTCTGCCCTGTTCACCACGCGCAACAAGAAAATCCCGCGCGAGGCGCTGCAAAACAAGGTCATTTTCCACGTCAACAAGGACGTGAAGATCACCTACACCGGCGTCGAGCTGCGGGCCGACGATGACGAACTGGTGTGGCAGCAAGTGTTGGAGTACGCGAAGCGCACCCCTATCGGCGAGCCGATCACCTTCACGTTCTACGAGCTTTGCCAGGACCTGGGCTGGTCGATCAATGGCCGGTATTACACGAAGGCCGAGGAATGCCTGTCGCGCCTACAGGCGACGGCCATGGGCTTCACGTCCGACCGCGTTGGGCACCTGGAATCGGTGTCGCTGCTGCACCGCTTCCGCGTCCTGGACCGTGGCAAGAAAACGTCCCGTTGCCAGGTCCTGATCGACGAGGAAATCGTCGTGCTGTTTGCTGGCGACCACTACACGAAATTCATATGGGAGAAGTACCGCAAGCTGTCGCCGACGGCCCGACGGATGTTCGACTATTTCAGCTCGCACCGGGAGCCGTACCCGCTCAAGCTGGAAACCTTCCGCCTCATGTGCGGATCGGATTCCACCCGCGTGAAGAAGTGGCGCGAGCAGGTCGGCGAAGCCTGCGAAGAGTTGCGAGGCAGCGGCCTGGTGGAACACGCCTGGGTCAATGATGACCTGGTGCATTGCAAACGCTAGGGCCTTGTGGGGTCAGTTCCGGCTGGGGGTTCAGCAGCCACCTG -pRO1600: - description: Broad host range plasmid (varies per host) for gram negative cells. - sequence: GCCGCTGGTGCCGCTGGTTGGACGCCAAGGGTGAATCCGCCTCGATACCCTGATTACTCGCTTCCTGCGCCCTCTCAGGCGGCGATAGGGGACTGGTAAAACGGGGATTGCCCAGACGCCTCCCCCGCCCCTTCAGGGGCACAAATGCGGCCCCAACGGGGCCACGTAGTGGTGCGTTTTTTGCGTTTCCACCCTTTTCTTCCTTTTCCCTTTTAAACCTTTTAGGACGTCTACAGGCCACGTAATCCGTGGCCTGTAGAGTTTAAAAAGGGACGGATTTGTTGCCATTAAGGGACGGATTTGTTGTTAAGAAGGGACGGATTTGTTGTTGTAAAGGGACGGATTTGTTGTATTGTGGGACGCAGATACAGTGTCCCCTTATACACAAGGAATGTCGAACGTGGCCTCACCCCCAATGGTTTACAAAAGCAATGCCCTGGTCGAGGCCGCGTATCGCCTCAGTGTTCAGGAACAGCGGATCGTTCTGGCCTGTATTAGCCAGGTGAAGAGGAGCGAGCCTGTCACCGATGAAGTGATGTATTCAGTGACGGCGGAGGACATAGCGACGATGGCGGGTGTCCCTATCGAATCTTCCTACAACCAGCTCAAAGAAGCGGCCCTGCGCCTGAAACGGCGGGAAGTCCGGTTAACCCAAGAGCCCAATGGCAAGGGGAAAAGACCGAGTGTGATGATTACCGGCTGGGTGCAAACAATCATCTACCGGGAGGGTGAGGGCCGTGTAGAACTCAGGTTCACCAAAGACATGCTGCCGTACCTGACGGAACTCACCAAACAGTTCACCAAATACGCCTTGGCTGACGTGGCCAAGATGGACAGCACCCACGCGATCAGGCTTTACGAGCTGCTCATGCAATGGGACAGCATCGGCCAGCGCGAAATAGAAATTGACCAGCTGCGAAAGTGGTTTCAACTGGAAGGCCGGTATCCCTCGATCAAGGACTTCAAGTTGCGAGTGCTTGATCCAGCCGTGACGCAGATCAACGAGCACAGCCCGCTACAGGTGGAGTGGGCGCAGCGAAAGACCGGGCGCAAGGTCACACATCTGTTGTTCAGTTTTGGACCGAAGAAGCCCGCCAAGGCGGTGGGTAAGGCCCCAGCGAAGCGCAAGGCCGGGAAGATTTCAGATGCTGAGATCGCGAAACAGGCTCGCCCTGGTGAGACATGGGAAGCGGCCCGCGCTCGACTAACCCAGATGCCGCTGGATCTGGCCTAGAGGCCGTGGCCACCACGGCCCGGCCTGCCTTTC -ColA: - description: Medium copy (20-40 copies) Escherichia coli origin. Compatible with pUC/pBR322, p15A, RSF1030, CloDF13, and SC101 - sequence: TGGTGTCGGGAATCCGTAAAGGATCTTCTTGAGATCCTTTTACGATCGTCGTAATCTCCTGCTCTGTAAACGAAAAAACCGCCTGGGGAGGCGGTTTGATCGAAGGTTAAGTCAGTTGGGGAACTGCTTAACCTGGTAACTGGCTTTAGTGGAGCGCAGATACCAAATACTGTCCTTTCAGTGTAGCCTCTGTTAGGCCACCACTTCAAGACTCTCGATATCTAAATCCACTAATTCTCAGTTACCAATGGCTGCTGCCAGTGGCGTTTTGTCGTGTCTTTCCGGGTTGGACTCAAGATGATAGTTACCGGATAAGGCGCAGCAGTCGGGCTGAACGGGGGGTTCTTGCACACAGCCCAGCTTGGAGCGAACTGTCTACACGGAACGGGACGTGGTGATTTGGGTAAAGCCTCCACCACAACACGGACGCCGCAGGACGGGAACAGGAGAGCGCAAGAGGGAGCCATCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCACTGATTTGAGCGTCAGATTTCGTGATGTTCGTCAGGGGGGCGGAGCCTATGGAAAAACGGCTTCGCTCCGGCCTTATTGTCTCTCTGCTAAGTATCCTCCTGGCATCTTCTAGGACGTTT -BAC: - description: Single copy origin built to create Bacterial Artificial Chromosomes, which allow for cloning and stable maintenance of very large DNA fragments (up to 300kb) - sequence: CATGGAGCGGCGTAACCGTCGCACAGGAAGGACAGAGAAAGCGCGGATCTGGGAAGTGACGGACAGAACGGTCAGGACCTGGATTGGGGAGGCGGTTGCCGCCGCTGCTGCTGACGGTGTGACGTTCTCTGTTCCGGTCACACCACATACGTTCCGCCATTCCTATGCGATGCACATGCTGTATGCCGGTATACCGCTGAAAGTTCTGCAAAGCCTGATGGGACATAAGTCCATCAGTTCAACGGAAGTCTACACGAAGGTTTTTGCGCTGGATGTGGCTGCCCGGCACCGGGTGCAGTTTGCGATGCCGGAGTCTGATGCGGTTGCGATGCTGAAACAATTATCCTGAGAATAAATGCCTTGGCCTTTATATGGAAATGTGGAACTGAGTGGATATGCTGTTTTTGTCTGTTAAACAGAGAAGCTGGCTGTTATCCACTGAGAAGCGAACGAAACAGTCGGGAAAATCTCCCATTATCGTAGAGATCCGCATTATTAATCTCAGGAGCCTGTGTAGCGTTTATAGGAAGTAGTGTTCTGTCATGATGCCTGCAAGCGGTAACGAAAACGATTTGAATATGCCTTCAGGAACAATAGAAATCTTCGTGCGGTGTTACGTTGAAGTGGAGCGGATTATGTCAGCAATGGACAGAACAACCTAATGAACACAGAACCATGATGTGGTCTGTCCTTTTACAGCCAGTAGTGCTCGCCGCAGTCGAGCGACAGGGCGAAGCCCTCGAGTGAGCGAGGAAGCACCAGGGAACAGCACTTATATATTCTGCTTACACACGATGCCTGAAAAAACTTCCCTTGGGGTTATCCACTTATCCACGGGGATATTTTTATAATTATTTTTTTTATAGTTTTTAGATCTTCTTTTTTAGAGCGCCTTGTAGGCCTTTATCCATGCTGGTTCTAGAGAAGGTGTTGTGACAAATTGCCCTTTCAGTGTGACAAATCACCCTCAAATGACAGTCCTGTCTGTGACAAATTGCCCTTAACCCTGTGACAAATTGCCCTCAGAAGAAGCTGTTTTTTCACAAAGTTATCCCTGCTTATTGACTCTTTTTTATTTAGTGTGACAATCTAAAAACTTGTCACACTTCACATGGATCTGTCATGGCGGAAACAGCGGTTATCAATCACAAGAAACGTAAAAATAGCCCGCGAATCGTCCAGTCAAACGACCTCACTGAGGCGGCATATAGTCTCTCCCGGGATCAAAAACGTATGCTGTATCTGTTCGTTGACCAGATCAGAAAATCTGATGGCACCCTACAGGAACATGACGGTATCTGCGAGATCCATGTTGCTAAATATGCTGAAATATTCGGATTGACCTCTGCGGAAGCCAGTAAGGATATACGGCAGGCATTGAAGAGTTTCGCGGGGAAGGAAGTGGTTTTTTATCGCCCTGAAGAGGATGCCGGCGATGAAAAAGGCTATGAATCTTTTCCTTGGTTTATCAAACGTGCGCACAGTCCATCCAGAGGGCTTTACAGTGTACATATCAACCCATATCTCATTCCCTTCTTTATCGGGTTACAGAACCGGTTTACGCAGTTTCGGCTTAGTGAAACAAAAGAAATCACCAATCCGTATGCCATGCGTTTATACGAATCCCTGTGTCAGTATCGTAAGCCGGATGGCTCAGGCATCGTGTCTCTGAAAATCGACTGGATCATAGAGCGTTACCAGCTGCCTCAAAGTTACCAGCGTATGCCTGACTTCCGCCGCCGCTTCCTGCAGGTCTGTGTTAATGAGATCAACAGCAGAACTCCAATGCGCCTCTCATACATTGAGAAAAAGAAAGGCCGCCAGACGACTCATATCGTATTTTCCTTCCGCGATATCACTTCCATGACGACAGGATAGTCTGAGGGTTATCTGTCACAGATTTGAGGGTGGTTCGTCACATTTGTTCTGACCTACTGAGGGTAATTTGTCACAGTTTTGCTGTTTCCTTCAGCCTGCATGGATTTTCTCATACTTTTTGAACTGTAATTTTTAAGGAAGCCAAATTTGAGGGCAGTTTGTCACAGTTGATTTCCTTCTCTTTCCCTTCGTCATGTGACCTGATATCGGGGGTTAGTTCGTCATCATTGATGAGGGTTGATTATCACAGTTTATTACTCTGAATTGGCTATCCGCGTGTGTACCTCTACCTGGAGTTTTTCCCACGGTGGATATTTCTTCTTGCGCTGAGCGTAAGAGCTATCTGACAGAACAGTTCTTCTTTGCTTCCTCGCCAGTTCGCTCGCTATGCTCGGTTACACGGCTGCGGCGAGCGCTAGTGATAATAAGTGACTGAGGTATGTGCTCTTCTTATCTCCTTTTGTAGTGTTGCTCTTATTTTAAACAACTTTGCGGTTTTTTGATGACTTTGCGATTTTGTTGTTGCTTTGCAGTAAATTGCAAGATTTAATAAAAAAACGCAAAGCAATGATTAAAGGATGTTCAGAATGAAACTCATGGAAACACTTAACCAGTGCATAAACGCTGGTCATGAAATGACGAAGGCTATCGCCATTGCACAGTTTAATGATGACAGCCCGGAAGCGAGGAAAATAACCCGGCGCTGGAGAATAGGTGAAGCAGCGGATTTAGTTGGGGTTTCTTCTCAGGCTATCAGAGATGCCGAGAAAGCAGGGCGACTACCGCACCCGGATATGGAAATTCGAGGACGGGTTGAGCAACGTGTTGGTTATACAATTGAACAAATTAATCATATGCGTGATGTGTTTGGTACGCGATTGCGACGTGCTGAGGACGTATTTCCACCGGTGATCGGGGTTGCTGCCCATAAAGGTGGCGTTTACAAAACCTCAGTTTCTGTTCATCTTGCTCAGGATCTGGCTCTGAAGGGGCTACGTGTTTTGCTCGTGGAAGGTAACGACCCCCAGGGAACAGCCTCAATGTATCACGGATGGGTACCAGATCTTCATATTCATGCAGAAGATACTCTCCTGCCTTTCTATCTTGGGGAAAAGGACGATGTCACTTATGCAATAAAGCCCACTTGCTGGCCGGGGCTTGACATTATTCCTTCCTGTCTGGCTCTGCACCGTATTGAAACTGAGTTAATGGGCAAATTTGATGAAGGTAAACTGCCCACCGATCCACACCTGATGCTCCGACTGGCCATTGAAACTGTTGCTCATGACTATGATGTCATAGTTATTGACAGCGCGCCTAACCTGGGTATCGGCACGATTAATGTCGTATGTGCTGCTGATGTGCTGATTGTTCCCACGCCTGCTGAGTTGTTTGACTACACCTCCGCACTGCAGTTTTTCGATATGCTTCGTGATCTGCTCAAGAACGTTGATCTTAAAGGGTTCGAGCCTGATGTACGTATTTTGCTTACCAAATACAGCAATAGTAATGGCTCTCAGTCCCCGTGGATGGAGGAGCAAATTCGGGATGCCTGGGGAAGCATGGTTCTAAAAAATGTTGTACGTGAAACGGATGAAGTTGGTAAAGGTCAGATCCGGATGAGAACTGTTTTTGAACAGGCCATTGATCAACGCTCTTCAACTGGTGCCTGGAGAAATGCTCTTTCTATTTGGGAACCTGTCTGCAATGAAATTTTCGATCGTCTGATTAAACCACGCTGGGAGATTAGATAATGAAGCGTGCGCCTGTTATTCCAAAACATACGCTCAATACTCAACCGGTTGAAGATACTTCGTTATCGACACCAGCTGCCCCGATGGTGGATTCGTTAATTGCGCGCGTAGGAGTAATGGCTCGCGGTAATGCCATTACTTTGCCTGTATGTGGTCGGGATGTGAAGTTTACTCTTGAAGTGCTCCGGGGTGATAGTGTTGAGAAAACCTCTCGGGTATGGTCAGGTAATGAACGTGACCAGGAGCTGCTTACTGAGGACGCACTGGATGATCTCATCCCTTCTTTTCTACTGACTGGTCAACAGACACCGGCGTTCGGTCGAAGAGTATCTGGTGTCATAGAAATTGCCGATGGGAGTCGCCGTCGTAAAGCTGCTGCACTTACCGAAAGTGATTATCGTGTTCTGGTTGGCGAGCTGGATGATGAGCAGATGGCTGCATTATCCAGATTGGGTAACGATTATCGCCCAACAAGTGCTTATGAACGTGGTCAGCGTTATGCAAGCCGATTGCAGAATGAATTTGCTGGAAATATTTCTGCGCTGGCTGATGCGGAAAATATTTCACGTAAGATTATTACCCGCTGTATCAACACCGCCAAATTGCCTAAATCAGTTGTTGCTCTTTTTTCTCACCCCGGTGAACTATCTGCCCGGTCAGGTGATGCACTTCAAAAAGCCTTTACAGATAAAGAGGAATTACTTAAGCAGCAGGCATCTAACCTTCATGAGCAGAAAAAAGCTGGGGTGATATTTGAAGCTGAAGAAGTTATCACTCTTTTAACTTCTGTGCTTAAAACGTCATCTGCATCAAGAACTAGTTTAAGCTCACGACATCAGTTTGCTCCTGGAGCGACAGTATTGTATAAGGGCGATAAAATGGTGCTTAACCTGGACAGGAGCCGTGTTCCAACTGAGTGTATAGAGAAAATTGAGGCCATTCTTAAGGAACTTGAAAAGCCAGCACCCTGATGCGACCACGTTTTAGTCTACGTTTATCTGTCTTTACTTAATGTCCTTTGTTACAGGCCAGAAAGCATAACTGGCCTGAATATTCTCTCTGGGCCCACTGTTCCACTTGTATCGTCGGTCTGATAATCAGACTGGGACCACGGTCCCACTCGTATCGTCGGTCTGATTATTAGTCTGGGACCACGGTCCCACTCGTATCGTCGGTCTGATTATTAGTCTGGGACCACGGTCCCACTCGTATCGTCGGTCTGATAATCAGACTGGGACCACGGTCCCACTCGTATCGTCGGTCTGATTATTAGTCTGGGACCATGGTCCCACTCGTATCGTCGGTCTGATTATTAGTCTGGGACCACGGTCCCACTCGTATCGTCGGTCTGATTATTAGTCTGGAACCACGGTCCCACTCGTATCGTCGGTCTGATTATTAGTCTGGGACCACGGTCCCACTCGTATCGTCGGTCTGATTATTAGTCTGGGACCACGATCCCACTCGTGTTGTCGGTCTGATTATCGGTCTGGGACCACGGTCCCACTTGTATTGTCGATCAGACTATCAGCGTGAGACTACGATTCCATCAATGCCTGTCAAGGGCAAGTATTGACATGTCGTCGTAACCTGTAGAACGGAGTAACCTCGGTGTGCGGTTGTATGCCTGCTGTGGATTGCTGCTGTGTCCTGCTTATCCACAACATTTTGCGCACGGTTATGTGGACAAAATACCTGGTTACC -R6K: - description: Medium copy (15-30 copies) or high copy depending on Pi expression. Requires Pi protein. Orthogonal to ColE2. - sequence: GATCTGAAGATCAGCAGTTCAACCTGTTGATAGTACGTACTAAGCTCTCATGTTTCACGTACTAAGCTCTCATGTTTAACGTACTAAGCTCTCATGTTTAACGAACTAAACCCTCATGGCTAACGTACTAAGCTCTCATGGCTAACGTACTAAGCTCTCATGTTTCACGTACTAAGCTCTCATGTTTGAACAATAAAATTAATATAAATCAGCAACTTAAATAGCCTCTAAGGTTTTAAGTTTTATAAGAAAAAAAAGAATATATAAGGCTTTTAAAGCCTTTAAGGTTTAACGGTTGTGGACAACAAGCCAGGGATGTAACGCACTGAGAAGCCCTTAGAGCCTCTCAAAGCAATTTTGAGTGACACAGGAACACTTAACGGCTGACATGG -CloDF13: - description: Medium copy (20-40 copies) Escherichia coli origin. Compatible with pUC/pBR322, p15A, RSF1030, ColA, and SC101 - sequence: GATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTTTTGCCCTGTAAACGAAAAAACCACCTGGGGAGGTGGTTTGATCGAAGGTTAAGTCAGTTGGGGAACTGCTTAACCGTGGTAACTGGCTTTCGCAGAGCACAGCAACCAAATCTGTCCTTCCAGTGTAGCCGGACTTTGGCGCACACTTCAAGAGCAACCGCGTGTTTAGCTAAACAAATCCTCTGCGAACTCCCAGTTACCAATGGCTGCTGCCAGTGGCGTTTTACCGTGCTTTTCCGGGTTGGACTCAAGTGAACAGTTACCGGATAAGGCGCAGCAGTCGGGCTGAACGGGGAGTTCTTGCTTACAGCCCAGCTTGGAGCGAACGACCTACACCGAGCCGAGATACCAGTGTGTGAGCTATGAGAAAGCGCCACACTTCCCGTAAGGGAGAAAGGCGGAACAGGTATCCGGTAAACGGCAGGGTCGGAACAGGAGAGCGCAAGAGGGAGCGACCCGCCGGAAACGGTGGGGATCTTTAAGTCCTGTCGGGTTTCGCCCGTACTGTCAGATTCATGGTTGAGCCTCACGGCTCCCACAGATGCACCGGAAAAGCGTCTGTTTATGTGAACTCTGGCAGGAGGGCGGAGCCTATGGAAAAACGCCACCGGCGCGGCCCTGCTGTTTTGCCTCACATGTTAGTCCCCTGCTTATCCACGGAATCTGTGGGTAACTTTGTATGTGTCCGCAGCGC -pLG13: - description: pLG13 is a ColE1 family plasmid that natively contains the EcoRV restriction-modification system - sequence: TTGAGATCCTTTTTTTTGCGCGCAATCTGCTGTCTGTAGACGAAAAAACCACCCTGGTAGGTGGTTTTTCGAAGGTTAAATAATCCTGGCAGATTATTTAACCGTGGTAACAGGGTGTACAAGACCGCTGCCACCAAATCTGTCCTTTCAGTGTAGCCGCAGTTGGTCCTTCACTTCAAGAACTACGTATCAGCAATTCTTGTACATCCTCTACCAGTGGCTGCTGCCAGTGGCGTTAAGGCGTGACTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCAACCCGAAGGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGGTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA -pSCANS-inducible: - description: Inducible copy number vector. Sourced from pOpen_v4 - sequence: agagcttggcaggtgtcgcgtatgcatcgatcacgtgtccacgttctttaatagtggactcttgttccaaactggaacaacactcggatcgatccggcgcgcaccgtgggaaaaactccaggtagaggtacacacgcggatagccaattcagagtaataaactgtgataatcaaccctcatcaatgatgacgaactaacccccgatatcaggtcacatgacgaagggaaagagaaggaaatcaactgtgacaaactgccctcaaatttggcttccttaaaaattacagttcaaaaagtatgagaaaatccatgcaggctgaaggaaacagcaaaactgtgacaaattaccctcagtaggtcagaacaaatgtgacgaaccaccctcaaatctgtgacagataaccctcagactatcctgtcgtcatggaagtgatatcgcggaaggaaaatacgatatgagtcgtctggcggcctttctttttctcaatgtatgagaggcgcattggagttctgctgttgatctcattaacacagacttgcaggaagcggcggcggaagtcaggcatacgctggtaactttgaggcagctggtaacgctctatgatccagtcgattttcagagacacgatgcctgagccatccggcttacgatactgacacagggattcgtataaacgcatggcatacggattggtgatttcttttgtttcactaagccgaaactgcgtaaaccggttctgtaacccgataaagaagggaatgagatatgggttgatatgtacactgtaaagccctctggatggactgtgcgcacgtttgataaaccaaggaaaagattcatagcctttttcatcaccggcatcctcttcagggcgataaaaaaccacttccttccccgcgaaactcttcaatgcctgccgtatatccttactggcttccgcagaggtcaatccgaatatttcagcatatttagcaacatggatctcgcagataccgtcatgttcctgtagggtgccatcagattttctgatctggtcaacgaacagatacagcatacgtttttgatcccgggagagactatatgccgcctcagtgaggtcgtttgactggacgattcgcgggctatttttacgtttcttgtgattgataaccgctgtttccgccatgacagatccatgtgaagtgtgacaagtttttagattgtcacactaaataaaaaagagtcaataagcagggataactttgtgaaaaaacagcttcttctgagggcaatttgtcacagggttaagggcaatttgtcacagacaggactgtcatttgagggtgatttgtcacactgaaagggcaatttgtcacaacaccttctgtagaaccagcatggataaaggcctacaaggcgctctaaaaaagaagatctaaaaactataaaaaaaataattataaaaatatccccgtggataagtggataaccccaagggaagttttttcaggcatcgtgtgtaagcagaatatataagtgctgttccctggtgcttcctcgctcactcgaaattcccggggatagctttatgcttgtaaaccgttttgtgaaaaaatttttaaaataaaaaaggggacctctagggtccccaattaattagtaatataatctattaaaggtcattcaaaaggtcatccaccggatcaattcccctgctcgcgcaggctgggtgccaagctctcgggtaacatcaaggcccgatccttggagcccttgccctcccgcacgatgatcgtgccgtgatcgaaatccagatccttgacccgcagttgcaaaccctcactgatccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatgtgagttagctcactcattaggcaccccaggctttacactttatgcttccggctcgtatgttgtgtggaattgtgagcggataacaatttcacacaggaaacagctatgaccatgattacgccaagcgcggtacctgtcgcggcaacgcgctaacagacgtagtaagaaccaccagcattgtaatgctggctaaagtcactttcctgagctgtataacgatgagcgattttactttttctggctatgaattggcctgctttgtaacacactccggtctatcccgtagcgccgggcatatcctgtcgcaatgtgcaaatctcgcggcaacaaccagtgaatacttcattcacaagcctcaccgcctgatcgcggcagaaactggttatagccaatcaaccgtcgttcgtgcattccgtgaagctgtaaacaaaggtatcctgtctgtagagattgttatcggcgatcaccgtgaacgtcgcgctaacctgtaccggtttacaccatcctttttggccttcgcacaacaagccaaaaatgcgctgatagaaagcaaattaaagatctcttcagcggcaaccaaggttaaagctgttctcgctaagacattggctttatttaattttttatccacacccccatgtcaaaatgataccccctccccctgtcaggatgacgtggcaataaagaataagaagtcacaagttaaaaaaacaaaaagatcagtttccggcggtgccggaacaaccagcctcaaaaaattgacttcatggatcgctaaggcaaaagcaaaggctgacaatctgcggttatccaaaaaacgcactcaaaaacatgagttcaagcagaaagtagaggcggctgcgcggaaatatgcttacctgaagaacaagcgttcgcctgatattggcgggatatcaaacttcgataacctaccgcattgcatgacggtaaacgaagctcttaatgcggttttagccaaaaataaagataacgaacaatggggtataccggcaggattcagagggtaatgaattgctctaattataaccatgcatactttcaacacctctagtttgccatgaggcaaactcataggtgtcctggtaagaggacactgttgccaaaactggacgcc diff --git a/parts/parts/ecoli/promoters.yaml b/parts/parts/ecoli/promoters.yaml deleted file mode 100644 index e6f5a49..0000000 --- a/parts/parts/ecoli/promoters.yaml +++ /dev/null @@ -1,101 +0,0 @@ -# Anderson -BBa_J23119: - prefix: TACA - suffix: AACT - tags: ['promoter', 'anderson_promoter'] - sequence: ttgacagctagctcagtcctaggtataatgctagc -BBa_J23100: - prefix: TACA - suffix: AACT - tags: ['promoter', 'anderson_promoter'] - sequence: ttgacggctagctcagtcctaggtacagtgctagc -BBa_J23101: - prefix: TACA - suffix: AACT - tags: ['promoter', 'anderson_promoter'] - sequence: tttacagctagctcagtcctaggtattatgctagc -BBa_J23102: - prefix: TACA - suffix: AACT - tags: ['promoter', 'anderson_promoter'] - sequence: ttgacagctagctcagtcctaggtactgtgctagc -BBa_J23103: - prefix: TACA - suffix: AACT - tags: ['promoter', 'anderson_promoter'] - sequence: ctgatagctagctcagtcctagggattatgctagc -BBa_J23104: - prefix: TACA - suffix: AACT - tags: ['promoter', 'anderson_promoter'] - sequence: ttgacagctagctcagtcctaggtattgtgctagc -BBa_J23105: - prefix: TACA - suffix: AACT - tags: ['promoter', 'anderson_promoter'] - sequence: tttacggctagctcagtcctaggtactatgctagc -BBa_J23106: - prefix: TACA - suffix: AACT - tags: ['promoter', 'anderson_promoter'] - sequence: tttacggctagctcagtcctaggtatagtgctagc -BBa_J23107: - prefix: TACA - suffix: AACT - tags: ['promoter', 'anderson_promoter'] - sequence: tttacggctagctcagccctaggtattatgctagc -BBa_J23108: - prefix: TACA - suffix: AACT - tags: ['promoter', 'anderson_promoter'] - sequence: ctgacagctagctcagtcctaggtataatgctagc -BBa_J23109: - prefix: TACA - suffix: AACT - tags: ['promoter', 'anderson_promoter'] - sequence: tttacagctagctcagtcctagggactgtgctagc -BBa_J23110: - prefix: TACA - suffix: AACT - tags: ['promoter', 'anderson_promoter'] - sequence: tttacggctagctcagtcctaggtacaatgctagc -BBa_J23111: - prefix: TACA - suffix: AACT - tags: ['promoter', 'anderson_promoter'] - sequence: ttgacggctagctcagtcctaggtatagtgctagc -BBa_J23112: - prefix: TACA - suffix: AACT - tags: ['promoter', 'anderson_promoter'] - sequence: ctgatagctagctcagtcctagggattatgctagc -BBa_J23113: - prefix: TACA - suffix: AACT - tags: ['promoter', 'anderson_promoter'] - sequence: ctgatggctagctcagtcctagggattatgctagc -BBa_J23114: - prefix: TACA - suffix: AACT - tags: ['promoter', 'anderson_promoter'] - sequence: tttatggctagctcagtcctaggtacaatgctagc -BBa_J23115: - prefix: TACA - suffix: AACT - tags: ['promoter', 'anderson_promoter'] - sequence: tttatagctagctcagcccttggtacaatgctagc -BBa_J23116: - prefix: TACA - suffix: AACT - tags: ['promoter', 'anderson_promoter'] - sequence: ttgacagctagctcagtcctagggactatgctagc -BBa_J23117: - prefix: TACA - suffix: AACT - tags: ['promoter', 'anderson_promoter'] - sequence: ttgacagctagctcagtcctagggattgtgctagc -BBa_J23118: - prefix: TACA - suffix: AACT - tags: ['promoter', 'anderson_promoter'] - sequence: ttgacggctagctcagtcctaggtattgtgctagc diff --git a/parts/parts/vnat.yaml b/parts/parts/vnat.yaml index 7664aba..000a629 100644 --- a/parts/parts/vnat.yaml +++ b/parts/parts/vnat.yaml @@ -1,178 +1,338 @@ #https://doi.org/10.1021/acssynbio.1c00126 -dummy_promoter: + +## Anderson +BBa_J23119: prefix: TACA suffix: AACT - sequence: N + tags: ['promoter', 'anderson_promoter'] + sequence: ttgacagctagctcagtcctaggtataatgctagc +BBa_J23100: + prefix: TACA + suffix: AACT + tags: ['promoter', 'anderson_promoter'] + sequence: ttgacggctagctcagtcctaggtacagtgctagc +BBa_J23101: + prefix: TACA + suffix: AACT + tags: ['promoter', 'anderson_promoter'] + sequence: tttacagctagctcagtcctaggtattatgctagc +BBa_J23102: + prefix: TACA + suffix: AACT + tags: ['promoter', 'anderson_promoter'] + sequence: ttgacagctagctcagtcctaggtactgtgctagc +BBa_J23103: + prefix: TACA + suffix: AACT + tags: ['promoter', 'anderson_promoter'] + sequence: ctgatagctagctcagtcctagggattatgctagc +BBa_J23104: + prefix: TACA + suffix: AACT + tags: ['promoter', 'anderson_promoter'] + sequence: ttgacagctagctcagtcctaggtattgtgctagc +BBa_J23105: + prefix: TACA + suffix: AACT + tags: ['promoter', 'anderson_promoter'] + sequence: tttacggctagctcagtcctaggtactatgctagc +BBa_J23106: + prefix: TACA + suffix: AACT + tags: ['promoter', 'anderson_promoter'] + sequence: tttacggctagctcagtcctaggtatagtgctagc +BBa_J23107: + prefix: TACA + suffix: AACT + tags: ['promoter', 'anderson_promoter'] + sequence: tttacggctagctcagccctaggtattatgctagc +BBa_J23108: + prefix: TACA + suffix: AACT + tags: ['promoter', 'anderson_promoter'] + sequence: ctgacagctagctcagtcctaggtataatgctagc +BBa_J23109: + prefix: TACA + suffix: AACT + tags: ['promoter', 'anderson_promoter'] + sequence: tttacagctagctcagtcctagggactgtgctagc +BBa_J23110: + prefix: TACA + suffix: AACT + tags: ['promoter', 'anderson_promoter'] + sequence: tttacggctagctcagtcctaggtacaatgctagc +BBa_J23111: + prefix: TACA + suffix: AACT + tags: ['promoter', 'anderson_promoter'] + sequence: ttgacggctagctcagtcctaggtatagtgctagc +BBa_J23112: + prefix: TACA + suffix: AACT + tags: ['promoter', 'anderson_promoter'] + sequence: ctgatagctagctcagtcctagggattatgctagc +BBa_J23113: + prefix: TACA + suffix: AACT + tags: ['promoter', 'anderson_promoter'] + sequence: ctgatggctagctcagtcctagggattatgctagc +BBa_J23114: + prefix: TACA + suffix: AACT + tags: ['promoter', 'anderson_promoter'] + sequence: tttatggctagctcagtcctaggtacaatgctagc +BBa_J23115: + prefix: TACA + suffix: AACT + tags: ['promoter', 'anderson_promoter'] + sequence: tttatagctagctcagcccttggtacaatgctagc +BBa_J23116: + prefix: TACA + suffix: AACT + tags: ['promoter', 'anderson_promoter'] + sequence: ttgacagctagctcagtcctagggactatgctagc +BBa_J23117: + prefix: TACA + suffix: AACT + tags: ['promoter', 'anderson_promoter'] + sequence: ttgacagctagctcagtcctagggattgtgctagc +BBa_J23118: + prefix: TACA + suffix: AACT + tags: ['promoter', 'anderson_promoter'] + sequence: ttgacggctagctcagtcctaggtattgtgctagc +# Non-Anderson +Vnat_dummy_promoter: + prefix: TACA + suffix: AACT + sequence: cccctggcgcccctt Ptrc_promoter: prefix: TACA suffix: AACT - sequence: N + sequence: gtctagggcggcggatttgtcctactcaggagagcgttcaccgacaaacaacagataaaacgaaaggcccagtctttcgactgagcctttcgttttatttgatgcagcgggtcgactcccctaggtcactgcccgctttccagtcgggaaacctgtcgtgccagctgcattaatgaatcggccaacgcgcggggagaggcggtttgcgtattgggcgccagggtggtttttcttttcaccagtgacacgggcaacagctgattgcccttcaccgcctggccctgagagagttgcagcaagcggtccacgctggtttgccccagcaggcgaaaatcctgtttgatggtggttaacggcgggatataacatgagctatcttcggtatcgtcgtatcccactaccgagatatccgcaccaacgcgcagcccggactcggtaatggcgcgcattgcgcccagcgccatctgatcgttggcaaccagcatcgcagtgggaacgatgccctcattcagcatttgcatggtttgttgaaaaccggacatggcactccagtcgccttcccgttccgctatcggctgaatttgattgcgagtgagatatttatgccagccagccagacgcagacgcgccgagacagaacttaatgggcccgctaacagcgcgatttgctggtgacccaatgcgaccagatgctccacgcccagtcgcgtaccatcttcatgggagaaaataatactgttgatgggtgtctggtcagagacatcaagaaataacgccggaacattagtgcaggcagcttccacagcaatggcatcctggtcatccagcggatagttaatgatcagcccactgacgcgttgcgcgagaagattgtgcaccgccgctttacaggcttcgacgccgcttcgttctaccatcgacaccaccacgctggcacccagttgatcggcgcgagatttaatcgccgcgacaatttgcgacggcgcgtgcagggccagactggaggtggcaacgccaatcagcaacgactgtttgcccgccagttgttgtgccacgcggttgggaatgtaattcagctccgccatcgccgcttccactttttcccgcgttttcgcagaaacgtggctggcctggttcaccacgcgggaaacggtctgataagagacaccggcatactctgcgacatcgtataacgttactggtttcacattcaccaccctgaattgactctcttccgggcgctatcatgccataccgcgaaaggttttgcgccaaagctttccctcgacaattcgataaatgtgagcggataacattgacattggtgagcggataacaagatactgagcacatcagcaggacgcactgacc Ptet_promoter: prefix: TACA suffix: AACT - sequence: N + sequence: ttttgttatcaataaaaaaggccccccgttagggaggcttattgttcgtcccatcacggaaaaaggttatgctgcttttaagacccactttcacatttaagttgtttttctaatccgcatatgatcaattcaaggccgaataagaaggctggctctgcaccttggtgatcaaataattcgatagcttgtcgtaataatggcggcatactatcagtagtaggtgtttccctttcttctttagcgacttgatgctcttgatcttccaatacgcaacctaaagtaaaatgccccacagcgctgagtgcatataatgcattctctagtgaaaaaccttgttggcataaaaaggctaattgattttcgagagtttcatactgtttttctgtaggccgtgtacctaaatgtacttttgctccatcgcgatgacttagtaaagcacatctaaaacttttagcgttattacgtaaaaaatcttgccagctttccccttctaaagggcaaaagtgagtatggtgcctatctaacatctcaatggctaaggcgtcgagcaaagcccgcttattttttacatgccaatacaatgtaggctgctctacacctagcctctgggcgagtttacgggttgttaaaccttcgattccgacctcattaagcagctctaatgcgctgctaatcactctacttttatctaatcgagacatcattaattcctaatttttgttgacactctatcattgatagagttattttaccactccctatcagtgatagagaaaagtgaa Para(Vn)_promoter: prefix: TACA suffix: AACT - sequence: N + sequence: tgttatccatccactggtagaggtgagtgttcgctatacatatttgtggatttagggacatttgttagtgacaaaaatcacagcggaaaaatgtagcgaatttgtccattcatttagccagtgtggctatgacacagatctcaattatgcgaccaatgatccaaattctcagtaagcaacccaataccagcctaatgcaaacgttgaattcgctggtttcttgcttttccgacctgacagaagaaggtgttaaaaagaacacaaaaaatcgtccatgacgtttttgtccataaagttagcagacctctttatgggataacatccctcctctagctataacaacaagtagatttagtttgctgaccaaaa Prham(Vn)_promoter: prefix: TACA suffix: AACT - sequence: N + sequence: gacacactctaataaccaagccccgcaattcgcggggcttattatttttagccagccaaatgttacgccctccccgttatttcaaacagtaaatagcttgaataaataagaaaaaccacaccttttacacgcctactccacttcacttaaacccaggttttatctggcctcacgcacgagttgtcaaaagtttgaattaccgcaagagcttcttgagaaaaacgcatcgaatacgttttttcagggggatttttgaagttatttagtgcggaaaaacggtgtaatacctctaaagaacaagaggtgttaatc # dropout sfGFP / mScarlet for each +Vnat_dummy_rbs: + prefix: AACT + suffix: AATG + sequence: agagtgtcaggatacccgataatc B0029_rbs: prefix: AACT suffix: AATG - sequence: N + sequence: agagttcacacaggaaacctaatc B0030_rbs: prefix: AACT suffix: AATG - sequence: N + sequence: agagattaaagaggagaaataatc B0031_rbs: prefix: AACT suffix: AATG - sequence: N + sequence: agagtcacacaggaaacctaatc B0032_rbs: prefix: AACT suffix: AATG - sequence: N + sequence: agagtcacacaggaaagtaatc B0033_rbs: prefix: AACT suffix: AATG - sequence: N + sequence: agagtcacacaggactaatc B0034_rbs: prefix: AACT suffix: AATG - sequence: N + sequence: agagaaagaggagaaataatc B0035_rbs: prefix: AACT suffix: AATG - sequence: N + sequence: agagattaaagaggagaataatc B0064_rbs: prefix: AACT suffix: AATG - sequence: N + sequence: agagaaagaggggaaataatc # cds lux_operon(Vn): prefix: A - suffix: ATCC - sequence: N + suffix: GGATCC + sequence: atgactaaaaaaatttcattcattattaacggccaggttgaaatctttcccgaaagtgatgatttagtgcaatccattaattttggtgataatagtgtttacctgccaatattgaatgactctcatgtaaaaaacattattgattgtaatggaaataacgaattacggttgcataacattgtcaattttctctatacggtagggcaaagatggaaaaatgaagaatactcaagacgcaggacatacattcgtgacttaaaaaaatatatgggatattcagaagaaatggctaagctagaggccaattggatatctatgattttatgttctaaaggcggcctttatgatgttgtagaaaatgaacttggttctcgccatatcatggatgaatggctacctcaggatgaaagttatgttcgggcttttccgaaaggtaaatctgtacatctgttggcaggtaatgttccattatctgggatcatgtctatattacgcgcaattttaactaagaatcagtgtattataaaaacatcgtcaaccgatccttttaccgctaatgcattagcgttaagttttattgatgtagaccctaatcatccgataacgcgctctttatctgttatatattggccccaccaaggtgatacatcactcgcaaaagaaattatgcgacatgcggatgttattgtcgcttggggagggccagatgcgattaattgggcggtagagcacgcgccatcttatgctgatgtgattaaatttggttctaaaaagagtctttgcattatcgataatcctgttgatttgacgtccgcagcgacaggtgcggctcatgatgtttgtttttacgatcagcgagcttgtttttctgcccaaaacatatattacatgggaaatcattatgaggaatttaagttagcgttgatagaaaaacttaatctatatgcgcatatattaccgaatgccaaaaaagattttgatgaaaaggcggcctattctttagttcaaaaagaaagcctgtttgctggattaaaagtagaggtggatattcatcaacgttggatgattattgagtcaaatgcaggtgtggaatttaatcaaccacttggcagatgtgtgtaccttcatcacgtcgataatattgagcaaatattgccttatgttcaaaaaaataagacgcaaaccatatctatttttccttgggagtcatcatttaaatatcgagatgcgttagcattaaaaggtgcggaaaggattgtagaagcaggaatgaataacatatttcgagttggtggatctcatgacggaatgaggccgttgcaacgattagtgacatatatttctcatgaaaggccatctaactatacggctaaggatgttgcggttgaaatagaacagactcgattcctggaagaagataagttccttgtatttgtcccataataggtaaaaagtatggaaaatgaatcaaaatataaaaccatcgaccacgttatttgtgttgaaggaaataaaaaaattcatgtttgggaaacgctgccagaagaaaacagcccaaagagaaagaatgccattattattgcgtctggttttgcccgcaggatggatcattttgctggtctggcggaatatttatcgcggaatggatttcatgtgatccgctatgattcgcttcaccatgttggcttgagttcagggacaattgatgaatttacaatgtctataggaaagcagagcttgttagcagtggttgattggttaactacacgaaaaataaataacttcggtatgttggcttccagcttatctgcgcggatagcttatgcaagcctatctgaaatcaatgcttcgtttttaatcaccgcagtcggtgttgttaacttaagatattctcttgaaagagctttagggtttgattatctcagtctacccattaatgaattgccggataatctggattttgaaggccataaattgggtgctgaagtctttgcgagagattgtcttgattttggttgggaagatttagcttctacaattaataacatgatgtatcttgatataccgtttattgcttttactgcaaataacgataattgggtcaagcaagatgaagttatcacattgttatcaaatattcgtagtaatcgatgcaagatatattctttgttaggaagttcgcatgacttgagtgaaaatttagtggtcctgcgcaatttttatcaatcggttacgaaagccgctatcgcgatggataatgatcatctggatattgatgttgatattactgaaccgtcatttgaacatttaactattgcgacagtcaatgaacgccgaatgagaattgagattgaaaatcaagcaatttctctgtcttaaaatctattgagatattctatcactcaaatagcaatataaggactctctatgaaatttggaaactttttgcttacataccaacctccccaattttctcaaacagaggtaatgaaacgtttggttaaattaggtcgcatctctgaggagtgtggttttgataccgtatggttactggagcatcatttcacggagtttggtttgcttggtaacccttatgtcgctgctgcatatttacttggcgcgactaaaaaattgaatgtaggaactgccgctattgttcttcccacagcccatccagtacgccaacttgaagatgtgaatttattggatcaaatgtcaaaaggacgatttcggtttggtatttgccgagggctttacaacaaggactttcgcgtattcggcacagatatgaataacagtcgcgccttagcggaatgctggtacgggctgataaagaatggcatgacagagggatatatggaagctgataatgaacatatcaagttccataaggtaaaagtaaaccccgcggcgtatagcagaggtggcgcaccggtttatgtggtggctgaatcagcttcgacgactgagtgggctgctcaatttggcctaccgatgatattaagttggattataaatactaacgaaaagaaagcacaacttgagctttataatgaagtggctcaagaatatgggcacgatattcataatatcgaccattgcttatcatatataacatctgtagatcatgactcaattaaagcgaaagagatttgccggaaatttctggggcattggtatgattcttatgtgaatgctacgactatttttgatgattcagaccaaacaagaggttatgatttcaataaagggcagtggcgtgactttgtattaaaaggacataaagatactaatcgccgtattgattacagttacgaaatcaatcccgtgggaacgccgcaggaatgtattgacataattcaaaaagacattgatgctacaggaatatcaaatatttgttgtggatttgaagctaatggaacagtagacgaaattattgcttccatgaagctcttccagtctgatgtcatgccatttcttaaagaaaaacaacgttcgctattatattagctaaggagaaagaaatgaaatttggattgttcttccttaacttcatcaattcaacaactgttcaagaacaaagtatagttcgtatgcaggaaataacggagtatgttgataagttgaattttgaacagattttagtgtatgaaaatcatttttcagataatggtgttgtcggcgctcctctgactgtttctggttttctgctcggtttaacagagaaaattaaaattggttcattaaatcacatcattacaactcatcatcctgtccgcatagcggaggaagcgtgcttattggatcagttaagtgaagggagatttattttagggtttagtgattgcgaaaaaaaagatgaaatgcatttttttaatcgcccggttgaatatcaacagcaactatttgaagagtgttatgaaatcattaacgatgctttaacaacaggctattgtaatccagataacgatttttatagcttccctaaaatatctgtaaatccccatgcttatacgccaggcggacctcggaaatatgtaacagcaaccagtcatcatattgttgagtgggcggccaaaaaaggtattcctctcatctttaagtgggatgattctaatgatgttagatatgaatatgctgaaagatataaagccgttgcggataaatatgacgttgacctatcagagatagaccatcagttaatgatattagttaactataacgaagatagtaataaagctaaacaagagactcgtgcatttattagtgattatgttcttgaaatgcaccctaatgaaaatttcgaaaataaacttgaagaaataattgcagaaaacgctgtcggaaattatacggagtgtataactgcggctaagttggcaattgaaaagtgtggtgcgaaaagtgtattgctgtcctttgaaccaatgaatgatttgatgagccaaaaaaatgtaatcaatattgttgatgataatattaagaagtaccacatggaatatacctaatagatttcgagttgcagcgaggcggcaagtgaacgaatccccaggagcatagataactatgtgactggggtgagtgaaagcagccaacaaagcagcagcttgaaagatgaagggtataaaagagtatgacagcagtgctgccatactttctaatattatcttgaggagtaaaacaggtatgacttcatatgttgataaacaagaaattacagcaagctcagaaattgatgatttgattttttcgagcgatccattagtgtggtcttacgacgagcaggaaaaaatcagaaagaaacttgtgcttgatgcatttcgtaatcattataaacattgtcgagaatatcgtcactactgtcaggcacacaaagtagatgacaatattacggaaattgatgacatacctgtattcccaacatcggtttttaagtttactcgcttattaacttctcaggaaaacgagattgaaagttggtttaccagtagcggcacgaatggtttaaaaagtcaggtggcgcgtgacagattaagtattgagagactcttaggctctgtgagttatggcatgaaatatgttggtagttggtttgatcatcaaatagaattagtcaatttgggaccagatagatttaatgctcataatatttggtttaaatatgttatgagtttggtggaattgttatatcctacgacatttaccgtaacagaagaacgaatagattttgttaaaacattgaatagtcttgaacgaataaaaaatcaagggaaagatctttgtcttattggttcgccatactttatttatttactctgccattatatgaaagataaaaaaatctcattttctggagataaaagcctttatatcataaccggaggcggctggaaaagttacgaaaaagaatctctgaaacgtgatgatttcaatcatcttttatttgatactttcaatctcagtgatattagtcagatccgagatatatttaatcaagttgaactcaacacttgtttctttgaggatgaaatgcagcgtaaacatgttccgccgtgggtatatgcgcgagcgcttgatcctgaaacgttgaaacctgtacctgatggaacgccggggttgatgagttatatggatgcgtcagcaaccagttatccagcatttattgttaccgatgatgtcgggataattagcagagaatatggtaagtatcccggcgtgctcgttgaaattttacgtcgcgtcaatacgaggacgcagaaagggtgtgctttgagcttaaccgaagcgtttgatagt azurite(Vn): prefix: A - suffix: ATCC - sequence: N + suffix: GGATCC + sequence: atgtctaaaggtgaagaactgtttaccggtgttgttccaattctggttgaactggatggtgatgttaacggtcacaaattttctgtttctggtgaaggcgaaggtgatgcaacctacggtaaactgaccctgaaatttatctgtaccactggtaaactgccagttccatggccaactctggttaccactctgtctcacggtgttcaatgtttttctcgttacccagatcacatgaaacagcacgattttttcaaaagcgcaatgccagaaggttacgttcaagaacgtaccatcttcttcaaagatgacggtaactacaaaacccgtgcggaagtgaaatttgaaggtgataccctggttaaccgtatcgaactgaaaggtatcgacttcaaagaggatggcaacattctgggtcacaaactggaatacaactttaactctcacaacatctacatcatggcggacaaacaaaaaaacggcatcaaagtgaacttcaagattcgccacaacatcgaagatggttctgttcaactggcagatcactaccaacaaaacaccccaattggtgatggtccagttctgctgccagataaccactacctgtctacccaatctgcactgtctaaagatccaaacgaaaaacgtgatcacatggtgctgctggaatttcgtaccgcagcaggtattacccacggtatggatgaactgtacaaa mTurquoise(Vn): prefix: A - suffix: ATCC - sequence: N + suffix: GGATCC + sequence: atggtttctaaaggtgaagaactgtttaccggtgttgttccaattctggttgaactggatggtgatgttaacggtcacaaattttctgtttctggtgaaggcgaaggtgatgcaacctacggtaaactgaccctgaaatttatctgtaccactggtaaactgccagttccatggccaactctggttaccactctgtcttggggtgttcaatgttttgcacgttacccagatcacatgaaacaacacgattttttcaaaagcgcaatgccagaaggttacgttcaagaacgtaccatcttcttcaaagatgacggtaactacaaaacccgtgcggaagtgaaatttgaaggtgataccctggttaaccgtatcgaactgaaaggtatcgacttcaaagaggatggcaacattctgggtcacaaactggaatacaactactttagcgataacgtgtacatcaccgcagataaacaaaaaaacggtatcaaggcgaacttcaaaatccgtcacaacattgaagatggtggtgttcaactggcagatcactaccaacaaaacactccaattggtgatggtccagttctgctgccagataaccactacctgtctacccaatctaaactgtctaaagacccaaacgaaaaacgtgatcacatggttctgctggaatttgttaccgcagcaggtattaccttaggtatggatgaactgtacaaa sfGFP(Vn): prefix: A - suffix: ATCC - sequence: N + suffix: GGATCC + sequence: atgcgtaaaggtgaagaactgtttaccggtgttgttccaattctggttgaactggatggtgatgttaacggtcacaaattttctgttcgtggtgaaggcgaaggtgatgcaaccaacggtaaactgaccctgaaatttatctgtaccactggtaaactgccagttccatggccaactctggttaccactctgacctacggtgttcaatgttttgcacgttacccagatcacatgaaacaacacgattttttcaaaagcgcaatgccagaaggttacgttcaagaacgtaccatctcttttaaagatgacggcacctacaaaacccgtgcggaagttaaatttgaaggtgataccctggttaaccgcattgaactgaaaggcatcgattttaaagaagatggtaacatcctgggccacaaactggaatacaactttaactctcacaacgtgtacatcaccgcagacaaacaaaaaaacggtatcaaagcgaacttcaagatccgtcacaacgttgaagatggttctgttcaactggcagatcactaccaacaaaacaccccaattggtgatggtccagttctgctgccagataaccactacctgtctacccaaagcgttctgtctaaagatccaaacgaaaaacgtgatcacatggtgctgctggaatttgttaccgcagcaggtattacccacggtatggatgaactgtacaaagca mVenus(Vn): prefix: A - suffix: ATCC - sequence: N + suffix: GGATCC + sequence: atggtttctaaaggtgaagaactgtttaccggtgttgttccaattctggttgaactggatggtgatgttaacggtcacaaattttctgtttctggtgaaggcgaaggtgatgcaacctacggtaaactgaccctgaaactgatttgtaccactggtaaactgccagttccatggccaactctggttaccaccttaggttacggtctgcaatgttttgcacgttacccagatcacatgaaacaacacgattttttcaaaagcgcaatgccagaaggttacgttcaagaacgtaccatcttcttcaaagatgacggtaactacaaaacccgtgcggaagtgaaatttgaaggtgataccctggttaaccgtatcgaactgaaaggtatcgacttcaaagaggatggcaacattctgggtcacaaactggaatacaactacaactctcacaacgtttacatcaccgcagataaacaaaaaaacggcatcaaagcgaacttcaaaatccgtcacaacattgaagatggtggtgttcaactggcagatcactaccaacaaaacaccccaattggtgatggtccagttctgctgccagataaccactacctgtcttaccaatctaaactgagcaaagacccaaacgaaaaacgtgatcacatggttctgctggaatttgttaccgcagcaggtattaccttaggtatggatgaactgtacaaa mScarlet-I(Vn): prefix: A - suffix: ATCC - sequence: N + suffix: GGATCC + sequence: atggtttctaaaggtgaagcagtgatcaaagaatttatgcgcttcaaagttcacatggaaggttctatgaacggccacgaatttgaaattgaaggtgaaggcgaaggtcgtccatacgaaggtactcaaaccgcaaaactgaaagttaccaaaggtggtccactgccattttcttgggatattctgtctccacaatttatgtacggttctcgtgcatttatcaaacacccagcagatattccagactactacaaacaatcttttccggaaggtttcaaatgggaacgtgttatgaattttgaagatggtggtgcagttacggttacccaagatacctctctggaagatggtactctgatctacaaagttaaactgcgtggtactaactttccaccagatggtccagttatgcagaaaaaaaccatgggttgggaagcatctaccgaacgtctgtacccagaagatggcgttctgaaaggtgatatcaaaatggcactgcgtctgaaagatggcggtcgttacctggcagatttcaaaaccacctacaaagcgaaaaaaccagttcaaatgccaggtgcatacaacgttgatcgtaaactggatattaccagccacaacgaagattacaccgttgttgaacaatacgaacgttctgaaggccgtcactctaccggtggtatggatgaactgtacaaa mCherry(Vn): prefix: A - suffix: ATCC - sequence: N + suffix: GGATCC + sequence: atggtttctaaaggtgaagaggataacatggcgatcatcaaagaatttatgcgcttcaaagttcacatggaaggttctgttaacggccacgaatttgaaattgaaggtgaaggcgaaggtcgtccatacgaaggtactcaaaccgcaaaactgaaagttaccaaaggtggtccactgccatttgcatgggatattctgtctccacagtttatgtacggtagcaaagcatacgttaaacacccagcagatattccagattacctgaaactgtcttttccggaaggtttcaaatgggaacgtgttatgaattttgaagatggtggtgttgttacggttacccaagattcttctctgcaagatggtgagtttatctacaaagttaaactgcgtggcaccaactttccatctgatggtccagttatgcagaaaaaaaccatgggttgggaagcatcttctgaacgtatgtacccagaagatggcgcactgaaaggtgaaattaaacaacgtctgaaacttaaagatggcggtcactacgatgcagaagttaaaaccacctacaaagcgaaaaaaccagttcaactgccaggtgcatacaacgttaacattaaactggatatcaccagccacaacgaagattacaccattgttgaacaatacgaacgtgcagaaggccgtcactctaccggtggtatggatgaactgtacaaa mKate-2(Vn): prefix: A - suffix: ATCC - sequence: N + suffix: GGATCC + sequence: atggtttctgaactgattaaagaaaacatgcacatgaaactgtacatggaaggtactgttaacaaccaccacttcaaatgtacctctgaaggtgaaggtaaaccatacgaaggtactcaaaccatgcgtattaaagcagttgaaggtggtccactgccatttgcatttgatattctggcaacctcttttatgtacggcagcaaaacctttatcaaccacactcaaggtatcccggattttttcaaacaaagctttccagaaggtttcacctgggaacgtgttaccacctacgaagatggtggtgttctgaccgcaactcaagatacctctctgcaagatggttgtctgatctacaacgttaaaatccgtggtgttaactttccatctaacggtccagttatgcagaaaaaaaccttaggttgggaagcatctaccgaaactctgtacccagcggatggtggtctggaaggtcgtgcagatatggcactgaaactggttggtggtggtcacctgatttgtaacctgaaaaccacctaccgttctaaaaaaccagcgaaaaatctgaaaatgccaggtgtttactacgttgatcgtcgtctggaacgtatcaaagaagcagataaagaaacctacgtggaacaacacgaagttgcagttgcacgttactgtgatctgccatctaaactgggtcaccgt # terminators -- make ctag versions as well B0010_terminator: prefix: ATCCTAA suffix: CGCT - sequence: N + sequence: ccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctc B0015_terminator: prefix: ATCCTAA suffix: CGCT - sequence: N + sequence: ccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttata B1002_terminator: prefix: ATCCTAA suffix: CGCT - sequence: N + sequence: cgcaaaaaaccccgcttcggcggggttttttcgc B1003_terminator: prefix: ATCCTAA suffix: CGCT - sequence: N + sequence: cgccaaaaaccccgcttcggcggggtttttccgc B1004_terminator: prefix: ATCCTAA suffix: CGCT - sequence: N + sequence: cgccgaaaaccccgcttcggcggggttttgccgc +B1005_terminator: + prefix: ATCCTAA + suffix: CGCT + sequence: cgccgcaaaccccgcttcggcggggtttcgccgc B1006_terminator: prefix: ATCCTAA suffix: CGCT - sequence: N + sequence: aaaaaaaaaccccgcccctgacagggcggggtttttttt B1007_terminator: prefix: ATCCTAA suffix: CGCT - sequence: N + sequence: cgcaaaaaaccccgcccctgacagggcggggttttttcgc B1009_terminator: prefix: ATCCTAA suffix: CGCT - sequence: N + sequence: cgccgaaaaccccgcccctgacagggcggggttttgccgc B1010_terminator: prefix: ATCCTAA suffix: CGCT - sequence: N + sequence: cgccgcaaaccccgcccctgacagggcggggtttcgccgc +Vnat_dummy_terminator: + prefix: ATCCTAA + suffix: CGCT + sequence: actcagttgtagtaacgagcggatagattcccagacccaccttcacgggcggtagcaggacctcaataataggattttcg +#ctag terminator +B0010_terminator_ctag: + prefix: ATGT + suffix: CGCT + sequence: ccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctc +B0015_terminator_ctag: + prefix: ATGT + suffix: CGCT + sequence: ccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttata +B1002_terminator_ctag: + prefix: ATGT + suffix: CGCT + sequence: cgcaaaaaaccccgcttcggcggggttttttcgc +B1003_terminator_ctag: + prefix: ATGT + suffix: CGCT + sequence: cgccaaaaaccccgcttcggcggggtttttccgc +B1004_terminator_ctag: + prefix: ATGT + suffix: CGCT + sequence: cgccgaaaaccccgcttcggcggggttttgccgc +B1005_terminator_ctag: + prefix: ATGT + suffix: CGCT + sequence: cgccgcaaaccccgcttcggcggggtttcgccgc +B1006_terminator_ctag: + prefix: ATGT + suffix: CGCT + sequence: aaaaaaaaaccccgcccctgacagggcggggtttttttt +B1007_terminator_ctag: + prefix: ATGT + suffix: CGCT + sequence: cgcaaaaaaccccgcccctgacagggcggggttttttcgc +B1009_terminator_ctag: + prefix: ATGT + suffix: CGCT + sequence: cgccgaaaaccccgcccctgacagggcggggttttgccgc +B1010_terminator_ctag: + prefix: ATGT + suffix: CGCT + sequence: cgccgcaaaccccgcccctgacagggcggggtttcgccgc +Vnat_dummy_terminator_ctag: + prefix: ATGT + suffix: CGCT + sequence: actcagttgtagtaacgagcggatagattcccagacccaccttcacgggcggtagcaggacctcaataataggattttcg # n and c tag # ntags: 3xFlag, 6xHis, Azurite, mTurqoise, sfGFP, mVenus, mScarlet-I, mCherry, mKate-2 3xFlag(Vn)_ctag: prefix: ATCC - suffix: ATGT - sequence: N + suffix: TAAATGT + sequence: gattataaggatcatgatggtgattataaggatcatgatatcgactacaaagacgatgacgacaag 6xHis(Vn)_ctag: prefix: ATCC - suffix: ATGT - sequence: N + suffix: TAAATGT + sequence: caccatcaccaccatcat Azurite(Vn)_ctag: prefix: ATCC - suffix: ATGT - sequence: N + suffix: TAAATGT + sequence: atgtctaaaggtgaagaactgtttaccggtgttgttccaattctggttgaactggatggtgatgttaacggtcacaaattttctgtttctggtgaaggcgaaggtgatgcaacctacggtaaactgaccctgaaatttatctgtaccactggtaaactgccagttccatggccaactctggttaccactctgtctcacggtgttcaatgtttttctcgttacccagatcacatgaaacagcacgattttttcaaaagcgcaatgccagaaggttacgttcaagaacgtaccatcttcttcaaagatgacggtaactacaaaacccgtgcggaagtgaaatttgaaggtgataccctggttaaccgtatcgaactgaaaggtatcgacttcaaagaggatggcaacattctgggtcacaaactggaatacaactttaactctcacaacatctacatcatggcggacaaacaaaaaaacggcatcaaagtgaacttcaagattcgccacaacatcgaagatggttctgttcaactggcagatcactaccaacaaaacaccccaattggtgatggtccagttctgctgccagataaccactacctgtctacccaatctgcactgtctaaagatccaaacgaaaaacgtgatcacatggtgctgctggaatttcgtaccgcagcaggtattacccacggtatggatgaactgtacaaa mTurquoise(Vn)_ctag: prefix: ATCC - suffix: ATGT - sequence: N + suffix: TAAATGT + sequence: atggtttctaaaggtgaagaactgtttaccggtgttgttccaattctggttgaactggatggtgatgttaacggtcacaaattttctgtttctggtgaaggcgaaggtgatgcaacctacggtaaactgaccctgaaatttatctgtaccactggtaaactgccagttccatggccaactctggttaccactctgtcttggggtgttcaatgttttgcacgttacccagatcacatgaaacaacacgattttttcaaaagcgcaatgccagaaggttacgttcaagaacgtaccatcttcttcaaagatgacggtaactacaaaacccgtgcggaagtgaaatttgaaggtgataccctggttaaccgtatcgaactgaaaggtatcgacttcaaagaggatggcaacattctgggtcacaaactggaatacaactactttagcgataacgtgtacatcaccgcagataaacaaaaaaacggtatcaaggcgaacttcaaaatccgtcacaacattgaagatggtggtgttcaactggcagatcactaccaacaaaacactccaattggtgatggtccagttctgctgccagataaccactacctgtctacccaatctaaactgtctaaagacccaaacgaaaaacgtgatcacatggttctgctggaatttgttaccgcagcaggtattaccttaggtatggatgaactgtacaaa sfGFP(Vn)_ctag: prefix: ATCC - suffix: ATGT - sequence: N + suffix: TAAATGT + sequence: atgcgtaaaggtgaagaactgtttaccggtgttgttccaattctggttgaactggatggtgatgttaacggtcacaaattttctgttcgtggtgaaggcgaaggtgatgcaaccaacggtaaactgaccctgaaatttatctgtaccactggtaaactgccagttccatggccaactctggttaccactctgacctacggtgttcaatgttttgcacgttacccagatcacatgaaacaacacgattttttcaaaagcgcaatgccagaaggttacgttcaagaacgtaccatctcttttaaagatgacggcacctacaaaacccgtgcggaagttaaatttgaaggtgataccctggttaaccgcattgaactgaaaggcatcgattttaaagaagatggtaacatcctgggccacaaactggaatacaactttaactctcacaacgtgtacatcaccgcagacaaacaaaaaaacggtatcaaagcgaacttcaagatccgtcacaacgttgaagatggttctgttcaactggcagatcactaccaacaaaacaccccaattggtgatggtccagttctgctgccagataaccactacctgtctacccaaagcgttctgtctaaagatccaaacgaaaaacgtgatcacatggtgctgctggaatttgttaccgcagcaggtattacccacggtatggatgaactgtacaaagca mVenus(Vn)_ctag: prefix: ATCC - suffix: ATGT - sequence: N + suffix: TAAATGT + sequence: atggtttctaaaggtgaagaactgtttaccggtgttgttccaattctggttgaactggatggtgatgttaacggtcacaaattttctgtttctggtgaaggcgaaggtgatgcaacctacggtaaactgaccctgaaactgatttgtaccactggtaaactgccagttccatggccaactctggttaccaccttaggttacggtctgcaatgttttgcacgttacccagatcacatgaaacaacacgattttttcaaaagcgcaatgccagaaggttacgttcaagaacgtaccatcttcttcaaagatgacggtaactacaaaacccgtgcggaagtgaaatttgaaggtgataccctggttaaccgtatcgaactgaaaggtatcgacttcaaagaggatggcaacattctgggtcacaaactggaatacaactacaactctcacaacgtttacatcaccgcagataaacaaaaaaacggcatcaaagcgaacttcaaaatccgtcacaacattgaagatggtggtgttcaactggcagatcactaccaacaaaacaccccaattggtgatggtccagttctgctgccagataaccactacctgtcttaccaatctaaactgagcaaagacccaaacgaaaaacgtgatcacatggttctgctggaatttgttaccgcagcaggtattaccttaggtatggatgaactgtacaaa mScarlet-I(Vn)_ctag: prefix: ATCC - suffix: ATGT - sequence: N + suffix: TAAATGT + sequence: atggtttctaaaggtgaagcagtgatcaaagaatttatgcgcttcaaagttcacatggaaggttctatgaacggccacgaatttgaaattgaaggtgaaggcgaaggtcgtccatacgaaggtactcaaaccgcaaaactgaaagttaccaaaggtggtccactgccattttcttgggatattctgtctccacaatttatgtacggttctcgtgcatttatcaaacacccagcagatattccagactactacaaacaatcttttccggaaggtttcaaatgggaacgtgttatgaattttgaagatggtggtgcagttacggttacccaagatacctctctggaagatggtactctgatctacaaagttaaactgcgtggtactaactttccaccagatggtccagttatgcagaaaaaaaccatgggttgggaagcatctaccgaacgtctgtacccagaagatggcgttctgaaaggtgatatcaaaatggcactgcgtctgaaagatggcggtcgttacctggcagatttcaaaaccacctacaaagcgaaaaaaccagttcaaatgccaggtgcatacaacgttgatcgtaaactggatattaccagccacaacgaagattacaccgttgttgaacaatacgaacgttctgaaggccgtcactctaccggtggtatggatgaactgtacaaa mCherry_ctag: prefix: ATCC - suffix: ATGT - sequence: N + suffix: TAAATGT + sequence: atggtttctaaaggtgaagaggataacatggcgatcatcaaagaatttatgcgcttcaaagttcacatggaaggttctgttaacggccacgaatttgaaattgaaggtgaaggcgaaggtcgtccatacgaaggtactcaaaccgcaaaactgaaagttaccaaaggtggtccactgccatttgcatgggatattctgtctccacagtttatgtacggtagcaaagcatacgttaaacacccagcagatattccagattacctgaaactgtcttttccggaaggtttcaaatgggaacgtgttatgaattttgaagatggtggtgttgttacggttacccaagattcttctctgcaagatggtgagtttatctacaaagttaaactgcgtggcaccaactttccatctgatggtccagttatgcagaaaaaaaccatgggttgggaagcatcttctgaacgtatgtacccagaagatggcgcactgaaaggtgaaattaaacaacgtctgaaacttaaagatggcggtcactacgatgcagaagttaaaaccacctacaaagcgaaaaaaccagttcaactgccaggtgcatacaacgttaacattaaactggatatcaccagccacaacgaagattacaccattgttgaacaatacgaacgtgcagaaggccgtcactctaccggtggtatggatgaactgtacaaa mKate-2_ctag: prefix: ATCC - suffix: ATGT - sequence: N + suffix: TAAATGT + sequence: atggtttctgaactgattaaagaaaacatgcacatgaaactgtacatggaaggtactgttaacaaccaccacttcaaatgtacctctgaaggtgaaggtaaaccatacgaaggtactcaaaccatgcgtattaaagcagttgaaggtggtccactgccatttgcatttgatattctggcaacctcttttatgtacggcagcaaaacctttatcaaccacactcaaggtatcccggattttttcaaacaaagctttccagaaggtttcacctgggaacgtgttaccacctacgaagatggtggtgttctgaccgcaactcaagatacctctctgcaagatggttgtctgatctacaacgttaaaatccgtggtgttaactttccatctaacggtccagttatgcagaaaaaaaccttaggttgggaagcatctaccgaaactctgtacccagcggatggtggtctggaaggtcgtgcagatatggcactgaaactggttggtggtggtcacctgatttgtaacctgaaaaccacctaccgttctaaaaaaccagcgaaaaatctgaaaatgccaggtgtttactacgttgatcgtcgtctggaacgtatcaaagaagcagataaagaaacctacgtggaacaacacgaagttgcagttgcacgttactgtgatctgccatctaaactgggtcaccgt I11012_ctag: prefix: ATCC - suffix: ATGT - sequence: N + suffix: TAAATGT + sequence: gcagcaaacgacgaaaactacgctgctgctgtt M0050_ctag: prefix: ATCC - suffix: ATGT - sequence: N + suffix: TAAATGT + sequence: gctgctaacgacgaaaactacgctctggctgct M0051_ctag: prefix: ATCC - suffix: ATGT - sequence: N + suffix: TAAATGT + sequence: gctgctaacgacgaaaactacaactacgctgacgcttct M0052_ctag: prefix: ATCC - suffix: ATGT - sequence: N + suffix: TAAATGT + sequence: gctgctaacgacgaaaactacgctgacgcttct From f5e97f3a422fd224d99bd63d7b128b8fda4aae49 Mon Sep 17 00:00:00 2001 From: Keoni Gandall Date: Tue, 15 Oct 2024 15:42:52 -0700 Subject: [PATCH 5/9] add linkers2 --- parts/linkers2.md | 40 ++++++++++++++++++++++++++++++++++++++++ 1 file changed, 40 insertions(+) create mode 100644 parts/linkers2.md diff --git a/parts/linkers2.md b/parts/linkers2.md new file mode 100644 index 0000000..f1f3e4b --- /dev/null +++ b/parts/linkers2.md @@ -0,0 +1,40 @@ +# DnaDesign Assembly + +DnaDesign Assembly (shortened as dd assembly) is a GoldenGate DNA assembly method similar to MoClo. The overhang set was changed from MoClo in order to accomodate the overhangs `GTCT` and `CGAG`, which are used in recursive DNA assemblies. The BsaI overhangs were optimized using [empirical data](https://doi.org/10.1371/journal.pone.0238592) from the paper "Enabling one-pot Golden Gate assemblies of unprecedented complexity using data-optimized assembly design". + +overhangs: `CGAG,GTCT,GGGG,AAAA,AACT,AATG,ATCC,CGCT,TTCT,AAGC,ATAG,ATTA,ATGT,ACTC,ACGA,TATC,TAGG,TACA,TTAC,TTGA,TGGA,GAAG,GACC,GCCG` +overhangs with numbers: +1. CGAG +2. TACA +3. AACT +4. AATG +5. ATCC +6. CGCT +7. GTCT +8. AAGC +9. ATAG +10. ATTA +11. TTCT +12. ATGT +13. ACTC +14. ACGA +15. TATC +16. TAGG +17. TACA +18. TTAC +19. TTGA +20. TGGA +21. GAAG +22. GACC +23. GCCG +24. AAAA +25. GGGG + +## Linkers +The most basic dd assembly will create a transcriptional unit. Oftentimes, however, you will want to be able to combine different transcriptional units together. In order to do these multi-level assemblies, we use linkers. Linkers are genetic parts that are used within an assembly reaction that define the part overhangs of the assembled construct, for use in the next assembly reaction. For example, we could have 3 genetic parts: + +* Promoter+RBS +* GFP +* Terminator + +While we could construct a simple transcriptional unit of `["Promoter+RBS", "GFP", "Terminator"]`, we might want to use this whole transcriptional unit in a different construct. In order to do that, we will add 2 linkers. The prefix linker (A) From b6260cfe7e3c6b3934ad6ffcae63b45cc4c720c2 Mon Sep 17 00:00:00 2001 From: Keoni Gandall Date: Tue, 15 Oct 2024 15:56:36 -0700 Subject: [PATCH 6/9] parts.csv for synthesis --- parts/make_csv.py | 31 ++++++++ parts/parts.csv | 191 ++++++++++++++++++++++++++++++++++++++++++++++ 2 files changed, 222 insertions(+) create mode 100644 parts/make_csv.py create mode 100644 parts/parts.csv diff --git a/parts/make_csv.py b/parts/make_csv.py new file mode 100644 index 0000000..18bf07e --- /dev/null +++ b/parts/make_csv.py @@ -0,0 +1,31 @@ +import os +import yaml +import csv +import sys + +def process_yaml_files(directory): + data = [] + for filename in os.listdir(directory): + if filename.endswith('.yaml'): + with open(os.path.join(directory, filename), 'r') as file: + yaml_data = yaml.safe_load(file) + for name, gene in yaml_data.items(): + sequence = gene['prefix'] + gene['sequence'].lower() + gene['suffix'] + data.append({ + 'name': name, + 'vector': 'pOpen_v3', + 'type': 'dna', + 'sequence': sequence + }) + return data + +def print_csv(data): + writer = csv.DictWriter(sys.stdout, fieldnames=['name', 'vector', 'type', 'sequence']) + writer.writeheader() + for row in data: + writer.writerow(row) + +if __name__ == '__main__': + directory = './parts' + yaml_data = process_yaml_files(directory) + print_csv(yaml_data) diff --git a/parts/parts.csv b/parts/parts.csv new file mode 100644 index 0000000..1ae9099 --- /dev/null +++ b/parts/parts.csv @@ -0,0 +1,191 @@ +name,vector,type,sequence +CUP1_promoter,pOpen_v3,dna,TACAatcttagccttgttgctagttagaaaaagacatttttgctgtcagtcactgtcaagagattcttttgctggcatttcttctaggagcaaaaagagcgatgcgtcttttccgctgaaccgttccagcaaaaaagactaccaacgcaatatggattgtcagaatcatataaaagagaagcaaataactccttgtcttgtatcaattgcattataatatcttcttgttagtgcaatatcatatagaagtcatcgaaatagatattaagaaaaacaaactgtacaatcaatcaatcaatcatcacataaaagatcAACT +GAL1_promoter,pOpen_v3,dna,TACAccccattatcttagcctaaaaaaaccttctctttggaactttcagtaatacgcttaactgctcattgctatattgaagtacggattagaagccgccgagcgggcgacagccctccgacggatgactctcctccgtgcgtcctcgtcatcaccggtcgcgttcctgaaacgcagatgtgcctcgcgccgcactgctccgaacaataaagattctacaatactagcttttatggttatgaagaggaaaaattggcagtaacctggccccacaaaccttcaaattaacgaatcaaattaacaaccataggatgataatgcgattagttttttagccttatttctggggtaattaatcagcgaagcgatgatttttgatctattaacagatatataaatggaaaagctgcataaccactttaactaatactttcaacattttcagtttgtattacttcttattcaaatgtcataaaagtatcaacaaaaaattgttaatatacctctatactttaacgtcaaggagaaaaaactataagatcAACT +MF(alpha)2_promoter,pOpen_v3,dna,TACAttgatctgccttttatagctaagaaaaaaaaagagaaaaaaataataagtgcgggtaacctcatgcattgtaaaacggtagggctacaaggcactacttactctacgctatagaagaagcttcaagacatggtgttgttcttatttgagcgaactcagctgtcaccgtttcgggaaactctatagttttctgcgtttcagtacgcagttgggcgtgctaaagttgttttcctaatttgctagtgttcacttgctcattgatgtccctaaaaggtaatgtcgcatagtaagatgtatgtcccatatcaaacgcattcctagaaaaacatgcactgaatgaataagcagttacttcgattactaatcttcacagcataatttttctcttatcttgtaaacctctttctatcttttcataaagtttgtcacttatatttcttcattggtacatcaatgccagcaacgatgtgcgcatctgggcgacgcctgtagtgattgttttcaaggtatcgagccaaactattcatcgttactgtttcaaatattcagttgtttcagtacagagtcgccgtggacctagtgaaacttggtgtctttacagcgcagagatgagggcttatatgtataaaagctgtccttgattctggtgtagtttgaggtgtccttcctatatctgtttttatattctatataatggataattactaccatcacctgcatcaaattccagtaaattcacatattggagaaaagatcAACT +MFA1_promoter,pOpen_v3,dna,TACAcgctagaaggaaattggaagaactaaatagagttctgaaaaacaatccatatttcaattcaaatgtgaacgtacacaattcgagactcaaagatgctgtaccgttcacgccgtttaacggtgatagagaagcacacccaaggtttacgttgaaaggttcagtatacaatgacccattcatcaaacatcttgagcacaggaaagaatttattgcgtctgggttcaacactaattatgcgtacgaaagggtgttgacagaggcatttatgggcttaggatgtgttatatccgaggagctttaaaacatcaggatagtgtgcaacgtggcataagctatgtaatcaactactttttattttctatgtacgcatatacatgcattcacgatctgtttcagtgttcagaaaaaaggcacctactgctacggttggcccatacctttattctttgttcttgttacaaacgagtgtgtaattacccaaaaaggaaatttacatgttaaatgaaacccagtaatcagaaaaaacagttaagaaacctaaaatggtagagataaagatacagattcagtggttgctgaaaatcaagtaaaaaaatgaaatagagtcatcatatataaaccgccagaaatgaattaatgagagggatctgtaactgtttctcggataaaaccaaaataagtacaaagccatcgaatagaaagatcAACT +REV1_promoter,pOpen_v3,dna,TACAgtgttgttatccgatacaaccggatatttttcttttaatgagtctaaaccgtgatagcttcaggttaatacaatcaaaaaaagctcaaatattcttttaatgccgcgttcacagattccaattgaatacaactaggtagttcattatatgaagcctttgctactatttttcactatagtctgccttcaccttaatgcagacatccacatattttaatcactttaaaataaaaaggaagatatattagaagctatgatccaatctgtaagccagattaaaattcacgaactcttctttcatttgaattgaatgctttgagttggggtagattatcgcaaattactcatcacatttattgactacgaacttgctgatgtcctttttttatttatatttttcttcagtgaagcgattttttttttacacagaccaagacggaaaaaagtagctaaggaagaaaacaaaatcatgaaaaaaatgtgaagtgatcatgcacatcgcatcaacttaaacattggcttagagatatatagagttagagtttacggcaacctttaagcaccaataccttttggcatagtctaaagacctggttcttaattttaaacaaatttaactaaagatttccctatcaaagaagtaacgagttgacagattttctcaaaataaatcgatactgcatttctaggcatatccagcgagatcAACT +PSP2_promoter,pOpen_v3,dna,TACAtgacccaacatcagatgacccaaggtccacctcttattaaaggacgtttgatccttcgacaccatggctctgttgaacttttatctgagagaggaaaaaaaggaaggaaaaaaaagaagaaacttcctttatttatttgtcttaaccacaacacacaatgcaataagatgcaatataatatcaaagccaatatcttatgttgctgatcctgagaaggaatatatacaatttatgtagtaaaataccttttcttctgcgagttgcaagaaatagaaaagactccgattgcgcatcgccagaataaaatttcacaaccacactttttggctgaactttttattacctgattaaacagagagagaaaaggtagaggtcaaaattttttaagcaaaactaaaaaagatgcaaaatcacgtgctgaaaatctaacataagggttaagattagagttttataggacttgttttgtaatatttcaaatacgagctaaccctactgatttcaattaggtctaatttagggttgagctgcactgaaatttcggaaattttgggttattttaaatgagacagaagaactacagagatacgttcttcagactttaaagcttatctccacaaagaattggtcaagaaatcatcctagaaaaacacgtttgctcactcgatcttaatcacatagagtgctggaacgggaagaaagatcAACT +RAD27_promoter,pOpen_v3,dna,TACAccttgtgaaattgcaaatatggtgatttgaaacgtttcctagtgcagcaggatcacagataacgtgtaaagggcttagcagttgataatcctctctagttaagacctaaacaaaatgctgtcactaaccgtagtattaaatgacacactttggtgactttcgttaatggggatgtggtagtggccattgccaataaacaaaaagaacagggaaagaagtagaaagtgatataagtttgcttgccacttttcgtttttcacgaaaaaaacaggcgaaaaaaaatgctagacaagtacccggctgaatcacacctcgttaacagtgactttcggtgacagatacccgattgggcacccggctggtaagttatgatagaaagccaacgctgtactattggcttagctatggcaatattttgattatcagctagttttattaacgttataattagtgtaaccagtttttcatctatttcatttatttcatttatttactttaattgcagatccccctaacgcgtttaaagcttttattcactagcttatgtattttttataggaaacgcgacgcgtaacatcgcgcaaatgaaggttttgatgtattataatgaggtattcttccttatatacatcgatgaaaagcgttgacagcatacattggaaagaaataggaaacggacaccggaagaaaaaatagatcAACT +POP6_promoter,pOpen_v3,dna,TACAttcgtgctttgtgataaagtgtttcacgtcatccgacatgacttcgtagttatggactgaactgtgtggtgaggttccatgatttcttaggtccagcagatacatgtctcttcccaatttcttgttaaggttacggccaatgcttcggttgttgagcttgttaccgaataagccgtgaagtatgataataggtggtcttggcttcccttcatccccagtttttactgcatctctcttgattatgtcatatgaaaggtccagtgggacttgcttttgttgcagcacctttgctaatgaatgaaaggcacatagtgactgcttaaaaatgcaggaacttaaattattccgaatggtattttgtctcacatatattgtcccatactgtgccaagatcccggctttacccagtatcatcattgtaccgttaccaattctcctcgtatatcacggttagtttttaaacctcggggtgacgtttactattggcgtactaatatattcttattttcttttcttttttgttggcagtttcaagcaacacatgtactggataaccaacccccgcacgctcttggaaaaaattgagaaggcatcggacacttgctgatgagtatttcgaaaaattccatgaagatgaggccaagattgtttggaagagattgaaaagaagaagaagaaaaaaagataaaagcaaatcaaaagatcAACT +RNR2_promoter,pOpen_v3,dna,TACAagtcgaacaagaagcaggcaaagtttagagcactgcccctccgcactcaaaaaagaaaaaactaggaggaaaataaaattctcaaccacacaaacacataaacacatacaaatacaaatacaagcttatttacttgacatcgcgcgatcttccactattcagcgccgtccgccctctctcgtgttttttgtttacgcgacaactatgcgaaatccggagcaacgggcaaccgtttggggaaagaccacacccacgcgcgatcgccatggcaacgaggtcgcacacgccccacacccagacctccctgcgagcgggcatgggtacaatgtccccgttgccacagacaccacttcgtagcacagcgcagagcgtagcgtgttgttgctgctgacaaaagaaaatttttcttagcaaagcaaaggaggggaagcacgggcagatagcaccgtaccatacccttggaaactcgaaatgaacgaagcaggaaatgagagaatgagagttttgtaggtatatatagcggtagtgtttgcgcgttaccatcatcttctggatctatctattgttcttttcctcatcactttcccctttttcgctcttcttcttgtcttttatttctttcttttttttaattgttccctcgattggctatctaccaaagaatccaaacttaatacacgtatttatttgtccaattaccagatcAACT +SAC6_promoter,pOpen_v3,dna,TACAtttgagaatgaccttccacgagctaaattgaaagggaagaatttattagttgaactcaagaaagaagaggatgacgtgggaaatggcatagaatcccttactaaatcgaacactaaactgaactccatgctggcgaacgaaggtaagatacacaaagctagtttccagaaaagtgtaaaatttaaactacctgataatatagtgactgaagaaaccgtggaacttaaagaaataaaggacttgctactacaaatgttgagatgacagcgagagattgaatcaagattatccaatatcgaacttcaactcacggaaataccgaaacataagtaatcatatcccttctcacattttttacacaggaagtaagcaagttatgttatatttccgacactataattaattcttagcagttaaaggtgctttgtctatattacatttacatacagcttgagtgatcctgaccggatatagggtcctattttcttacgtgaacggcttttcttcttgttcccgatggccttcatgtgaaaaagcactcctcgggaggcggaaaaatatcaaaagtacggggcgaagtttataatgaagatttatcgatataaattttggttatttcaggagaacaagaaagctctttacactaaaattatcagagaagaagctgatatattagccctaaggagtacaccaaaacacaagatcAACT +RNR1_promoter,pOpen_v3,dna,TACAggatatcgtaaacaaaggcgttaccatagaaatgtactgattggcagaattactcttcaggagaatctttcatacaaaggtattccattggggaaaatctcgttaccaagtcaatgctgaactttctatggcctttgtttactatcgttaattattttacgaccacttctgggtagaaatatttcgtagccctggaacgagcttgtttacgcgttttatcccattatatggcacccaaatcaaatttaaaaagaaaaaacgcgtaaacagtgtcgggtaagttcatcctctgttactttaattgcttctttttttgaaattctaagtaaacgcgtcattttgatcctcaggacacagaaatccttgcagaatcttattgggtgttgaatagaggacgcgtaaaaacgatatggaaatttttttcatatagtgtagaaagaataggttggcgtaggtagtttcgtgtttgatagaaacctccaacaaagtctgcaactcacgttttagaataacaagtttagagtttatcttgttgcctttgttaagtcagtaccattgaataaaaattatataaaggagctaatatttcattgttggaaaattactctaccataattgaagcatatctcatccttttcatccttttcaacgcaagagagacaccaacgaacaacactttatttgttgatatattaacatcagatcAACT +RET2_promoter,pOpen_v3,dna,TACAacgatggcttcttatctcacttcaatagtactttccaccggttatacttccggcttttccctattaatacaagctacaatttcaatgggtggcaaataatgtgtagaatagaaaataagccgacagggtaataaagaaaatttttagaaaaaaaaggttagatggcttatttaagttacaggctagcgaaaaaaggaacttcagggcaagtaaagtgtttgattgggcactagcatggcttataaaggcgagcaattgtcgaaactaattaatgttgtacggactattgctgtcatctcgtggtaaatgcgtgttccaggtcgaatactacttgcacacaggcgagcggggccccataaaagtgttgccgatttgttaagttgtcttttcggtttttctactctgttattccttacttccctttttaagaactctttttatccttcatttaggatcttgcacgtttccgcctcatcacttgaattaaaacatgtctctgtcagtaaaccttggcgtttctattgttcttcatagttcaacttttattattacccgccctgcgcgtttacatttttccagcaacagccagcgaaaaattagaaaatctggttgttgacacctcaagaacaagggcaattagcctcagcgtcgaatatagatcatattagaatacctatagctccatcaaaagaaatacacaagatcAACT +PAB1_promoter,pOpen_v3,dna,TACAaaggcaagcccagaaaaatatcgcaagcacctttggtcttacagtgccaacttttggcctgccgacgttaagagtacaaagctgatggcaatgtacgacaagataacagagtctcaaaagaagtgaaacaatttttcttcaccacattttccattgttccttccccccataactataaacgtatttatgtatatatatttgcgtgtaagtgtgtgtactatagggcaccgtaaagtaataatgcttaattagttactactatgaccatataagaggtcatactgtatgaagccacaaagcagatagatcaatcatgtttaacgaaaactgttaatcgaagattatttctttttttttttctctttcctttttacaaagaaaattttttttgcgctttttgccatcaccatcgcaagttctgggacaattgttctctttcgctccagttccaaggaaagaggtttctgttttacttaatagaaagtgtcatcttgtattttatatctcttctttcttgtgtaaaattctttagttttgattttgtatttttaggacagtgagctacgaagtaacatttttacttaataaccgtttgaagcatagagcaggccctggtatcaccacctaatatctggctttttattcaataaaaactcaaaaaaaaaaatccaaaaaaaactaaaaaaccaataaaaataaaagatcAACT +ALD6_promoter,pOpen_v3,dna,TACAtaagggcatgatagaattggattatgtaaaaggtgaagataccattgtagaagcaaccagcacgtcgccgtggctgatgaagtctcctcttgcccgggccgcagaaaagaggggcagtggcctgtttttcgacataaatgaggggcatggccagcaccaagacgtcattgttgcatatggcgtatccaagccgaaacggcgctcgcctcatccccacgggaataaggcagccgacaaaagaaaaacgaccgaaaaggaaccagaaagaaaaaagagggtgggcgcgccgcggacgtgtaaaaagatatgcatccagcttctatatcgctttaactttaccgttttgggcatcgggaacgtatgtaacattgatctcctcttgggaacggtgagtgcaacgaatgcgatatagcaccgaccatgtgggcaaattcgtaataaattcggggtgagggggattcaagacaagcaaccttgttagtcagctcaaacagcgatttaacggttgagtaacacatcaaaacaccgttcgaggtcaagcctggcgtgtttaacaagttcttgatatcatatataaatgtaataagaagtttggtaatattcaattcgaagtgttcagtcttttacttctcttgttttatagaagaaaaaacatcaagaaacatctttaacatacacaaacacatactatcagaatacaagatcAACT +RPL18B_promoter,pOpen_v3,dna,TACAaagaggatgtccaatattttttttaaggaataaggatacttcaagactagattcccccctgcattcccatcagaaccgtaaaccttggcgctttccttgggaagtattcaagaagtgccttgtccggtttctgtggctcacaaaccagcgcgcccgatatggctttcttttcacttatgaatgtaccagtacgggacaattagaacgctcctgtaacaatctctttgcaaatgtggggttacattctaaccatgtcacactgctgacgaaattcaaagtaaaaaaaaatgggaccacgtcttgagaacgatagattttctttattttacattgaacagtcgttgtctcagcgcgctttatgttttcattcatacttcatattataaaataacaaaagaagaatttcatattcacgcccaagaaatcaggctgctttccaaatgcaattgacacttcattagccatcacacaaaactctttcttgctggagcttcttttaaaaaagacctcagtacaccaaacacgttacccgacctcgttattttacgacaactatgataaaattctgaagaaaaaataaaaaaattttcatacttcttgcttttatttaaaccattgaatgatttcttttgaacaaaactacctgtttcaccaaaggaaatagaaagaaaaaatcaattagaagaaaacaaaaaacaaaagatcAACT +HTB2_promoter,pOpen_v3,dna,TACAtatatattaaatttgctcttgttctgtactttcctaattcttatgtaaaaagacaagaatttatgatactatttaataacaaaaaactacctaagaaaagcatcatgcagtcgaaattgaaatcgaaaagtaaaactttaacggaacatgtttgaaattctaagaaagcatacatcttcatcccttatatatagagttatgtttgatattagtagtcatgttgtaatctctggcctaagtatacgtaacgaaaatggtagcacgtcgcgtttatggcccccaggttaatgtgttctctgaaattcgcatcactttgagaaataatgggaacaccttacgcgtgagctgtgcccaccgcttcgcctaataaagcggtgttctcaaaatttctccccgttttcaggatcacgagcgccatctagttctggtaaaatcgcgcttacaagaacaaagaaaagaaacatcgcgtaatgcaacagtgagacacttgccgtcatatataaggttttggatcagtaaccgttatttgagcataacacaggtttttaaatatattattatatatcatggtatatgtgtaaaatttttttgctgactggttttgtttatttatttagctttttaaaaattttactttcttcttgttaattttttctgattgctctatactcaaaccaacaacaacttactctacaactaagatcAACT +HHF1_promoter,pOpen_v3,dna,TACAtcttggggccttaccaccagtggactttcttgctgtttgctttgttctggccattgtttgcgtttatatatttatgttagatgtttttcttattaactagaaagaaagaatataaaaggttgaggaaagagatgtatcccgaagaatacacagtcttttatatatgtatttcaacaaggagccgtggagggtactaaaaagaaaaatcgcccgggcatttcgttatcttccacgctaaaagtcaaggagagatattacggccaggatcgcaaaggtgcagagcaaggaaatgtgagaaattgtgagaacgataatgtatgggacaatgcgaaaatgtgagaacgagagcaaaaatcttttttgtatctccccgccgaatttggaaaccgcgttctgaaaacttcgcatcttcacatagtaaaactgttccgagcgcttctccccataatggttagtggtaaaaaccgaagttgtttactttagcaaatgcccgcgaatacggtggtaaattgccacccccccttccccattcattgggtaaagaccaatttgatggataaattggttgtggaaaaggtctaattctttttcctataaataccgagatattttttctatatgatggtttccgtcgcattattgtactctatagtactaaagcaacaaacaaaaacaagcaacaaatataatatagtaaaatagatcAACT +TEF2_promoter,pOpen_v3,dna,TACAttgataggtcaagatcaatgtaaacaattactttgttatgtagagtttttttagctacctatattccaccataacatcaatcatgcggttgctggtgtatttaccaataatgtttaatgtatatatatatatatatatatggggccgtatacttacatatagtagatgtcaagcgtaggcgcttcccctgccggctgtgagggcgccataaccaaggtatctatagaccgccaatcagcaaactacctccgtacattcatgttgcacccacacatttatacacccagaccgcgacaaattacccataaggttgtttgtgacggcgtcgtacaagagaacgtgggaactttttaggctcaccaaaaaagaaagaaaaaatacgagttgctgacagaagcctcaagaaaaaaaaaattcttcttcgactatgctggaggcagagatgatcgagccggtagttaactatatatagctaaattggttccatcaccttcttttctggtgtcgctccttctagtgctatttctggcttttcctatttttttttttccatttttctttctctctttctaatatataaattctcttgcattttctatttttctctctatctattctacttgtttattcccttcaaggtttttttttaaggagtacttgtttttagaatatacggtcaacgaactataattaactaaacagatcAACT +TEF1_promoter,pOpen_v3,dna,TACAccttgccaacagggagttcttcagagacatggaggctcaaaacgaaattattgacagcctagacatcaatagtcatacaacagaaagcgaccacccaactttggctgataatagcgtataaacaatgcatactttgtacgttcaaaatacaatgcagtagatatatttatgcatattacatataatacatatcacataggaagcaacaggcgcgttggacttttaattttcgaggaccgcgaatccttacatcacacccaatcccccacaagtgatcccccacacaccatagcttcaaaatgtttctactccttttttactcttccagattttctcggactccgcgcatcgccgtaccacttcaaaacacccaagcacagcatactaaatttcccctctttcttcctctagggtgtcgttaattacccgtactaaaggtttggaaaagaaaaaagacaccgcctcgtttctttttcttcgtcgaaaaaggcaataaaaatttttatcacgtttctttttcttgaaaatttttttttttgatttttttctctttcgatgacctcccattgatatttaagttaataaacggtcatcaatttctcaagtttcagtttcatttttcttgttctattacaactttttttacttcttgctcattagaaagaaagcatagcaatctaatctaagttttaattacaaaagatcAACT +HHF2_promoter,pOpen_v3,dna,TACAtgtggagtgtttgcttggattctttagtaaaaggggaagaacagttggaagggccaaagtggaagtcacaaaacagtggtcctatataaaagaacaagaaaaagattatttatatacaactgcggtcacaagaagcaacgcgagagagcacaacacgctgttatcacgcaaactatgttttgacaccgagccatagccgtgattgtgcgtcacattgggcgataatgaacgctaaatgaccaactcccatccgtaggagccccttagggcgtgccaatagtttcacgcgcttaatgcgaagtgctcggaacggacaactgtggtcgtttggcaccgggaaagtggtactagaccgagagtttcgcatttgtatggcaggacgttctgggagcttcgcgtctaaagctttttcgggcgcgaaatgcagaccagaccagaacaaaacaactgacaagaaggcgtttaatttaatatgttgttcactcgcgcctgggctgttgttattcggctagatacatacgtgtttgtgcgtatgtagttatatcatatataagtatattaggatgaggcggtgaaagagattttttttttttcgcttaatttattcttttctctatcttttttcctacatcttgttcaaaagagtagcaaaaacaacaatcaatacaataaaataagatcAACT +PGK1_promoter,pOpen_v3,dna,TACAgtgagtaaggaaagagtgaggaactatcgcatacctgcatttaaagatgccgatttgggcgcgaatcctttattttggcttcaccctcatactattatcagggccagaaaaaggaagtgtttccctccttcttgaattgatgttaccctcataaagcacgtggcctcttatcgagaaagaaattaccgtcgctcgtgatttgtttgcaaaaagaacaaaactgaaaaaacccagacacgctcgacttcctgtcatcctattgattgcagcttccaatttcgtcacacaacaaggtcctagcgacggctcacaggttttgtaacaagcaatcgaaggttctggaatggcgggaaagggtttagtaccacatgctatgatgcccactgtgatctccagagcaaagttcgttcgatcgtactgttactctctctctttcaaacagaattgtccgaatcgtgtgacaacaacagcctgttctcacacactcttttcttctaaccaagggggtggtttagtttagtagaacctcgtgaaacttacatttacatatatataaacttgcataaattggtcaatgcaagaaatacatatttggtcttttctaattcgtagtttttcaagttcttagatgctttctttttctcttttttacagatcatcaaggaagtaattatctactttttacaacaaatataaaacaagatcAACT +CCW12_promoter,pOpen_v3,dna,TACAcacccatgaaccacacggttagtccaaaaggggcagttcagattccagatgcgggaattagcttgctgccaccctcacctcactaacgctgcggtgtgcggatacttcatgctatttatagacgcgcgtgtcggaatcagcacgcgcaagaaccaaatgggaaaatcggaatgggtccagaactgctttgagtgctggctattggcgtctgatttccgttttgggaatcctttgccgcgcgcccctctcaaaactccgcacaagtcccagaaagcgggaaagaaataaaacgccaccaaaaaaaaaaaaataaaagccaatcctcgaagcgtgggtggtaggccctggattatcccgtacaagtatttctcaggagtaaaaaaaccgtttgttttggaatttcccatttcgcggccacctacgccgctatctttgcaacaactatctgcgataactcagcaaattttgcatattcgtgttgcagtattgcgataatgggagtcttacttccaacataacggcagaaagaaatgtgagaaaattttgcatcctttgcctccgttcaagtatataaagtcggcatgcttgataatctttctttccatcctacattgttctaattattcttattctcctttattctttcctaacataccaagaaattaatcttctgtcattcgcttaaacactatatcaataaagatcAACT +TDH3_promoter,pOpen_v3,dna,TACAcagttcgagtttatcattatcaatactgccatttcaaagaatacgtaaataattaatagtagtgattttcctaactttatttagtcaaaaaattagccttttaattctgctgtaacccgtacatgcccaaaatagggggcgggttacacagaatatataacatcgtaggtgtctgggtgaacagtttattcctggcatccactaaatataatggagcccgctttttaagctggcatccagaaaaaaaaagaatcccagcaccaaaatattgttttcttcaccaaccatcagttcataggtccattctcttagcgcaactacagagaacaggggcacaaacaggcaaaaaacgggcacaacctcaatggagtgatgcaacctgcctggagtaaatgatgacacaaggcaattgacccacgcatgtatctatctcattttcttacaccttctattaccttctgctctctctgatttggaaaaagctgaaaaaaaaggttgaaaccagttccctgaaattattcccctacttgactaataagtatataaagacggtaggtattgattgtaattctgtaaatctatttcttaaacttcttaaattctacttttatagttagtcttttttttagttttaaaacaccaagaacttagtttcgaataaacacacataaacaaacaaaagatcAACT +SceI(Scerevisae),pOpen_v3,dna,AatgaagaacatcaagaagaaccaagttatgaacttgggtccaaactctaagttgttgaaggaatacaagtctcaattgatcgaattgaacatcgaacaattcgaagctggtatcggtttgatcttgggtgacgcttacatccgatctcgcgacgaaggtaagacctactgtatgcaattcgaatggaagaacaaggcttacatggaccacgtttgtttgttgtacgaccaatgggttttgtctccaccacacaagaaggaaagagttaaccacttgggtaacttggttatcacctggggtgctcaaaccttcaagcaccaagctttcaacaagttggctaacttgttcatcgttaacaacaagaaaaccatcccaaacaacttggttgaaaactacttgaccccaatgtctttggcttactggttcatggacgacggtggtaagtgggactacaacaagaactctaccaacaagtctatcgttttgaacacccaatctttcaccttcgaagaagttgaatacttggttaagggtttgagaaacaagttccaattgaactgttacgttaagatcaacaagaacaagccaatcatctacatcgactctatgtcttacttgatcttctacaacttgatcaagccatacttgatcccacaaatgatgtacaagttgccaaacaccatctcttctgaaaccttcttgaagGGATCC +mRuby2(Scerevisae),pOpen_v3,dna,AatggtgtccaaaggagaggagttaatcaaggaaaacatgagaatgaaagttgtcatggagggctccgttaatggtcaccaattcaagtgtacaggggaaggtgaaggtaatccttacatgggtacacaaactatgagaattaaagtaattgaaggcggaccactaccatttgcatttgacattctggcaacgtcattcatgtacggatcacgaactttcatcaagtaccctaaaggtataccagactttttcaagcaatcttttccagagggttttacatgggaaagggttacaagatacgaagatgggggtgtcgtcacagttatgcaagatacttcattagaagatggctgccttgtctatcatgtgcaagtaagaggggtgaattttccttctaacggacctgtgatgcagaaaaagaccaaaggttgggaaccaaatactgaaatgatgtacccagctgatggaggtttgagaggctacacacacatggcgcttaaagttgatggtggaggtcatttgtcttgtagttttgttaccacttatcgttctaaaaagactgttggcaatatcaaaatgccaggaatacatgctgtagaccacagactagaaagactcgaagagagcgataacgaaatgttcgttgtacagagagagcatgccgtagccaaatttgctggcttaggcggtggtatggatgaattgtataagGGATCC +Venus(Scerevisae),pOpen_v3,dna,AatgtctaaaggtgaagaattattcactggtgttgtcccaattttggttgaattagatggtgatgttaatggtcacaaattttctgtctccggtgaaggtgaaggtgatgctacttacggtaaattgaccttaaaattgatttgtactactggtaaattgccagttccatggccaaccttagtcactactttaggttatggtttgcaatgttttgctagatacccagatcatatgaaacaacatgactttttcaagtctgccatgccagaaggttatgttcaagaaagaactatttttttcaaagatgacggtaactacaagaccagagctgaagtcaagtttgaaggtgataccttagttaatagaatcgaattaaaaggtattgattttaaagaaggtggtaacattttaggtcacaaattggaatacaactataactctcacaatgtttacatcactgctgacaaacaaaagaatggtatcaaagctaacttcaaaattagacacaacattgaagatggtggtgttcaattagctgaccattatcaacaaaatactccaattggtgatggtccagtcttgttaccagacaaccattacttatcctatcaatctgccttatccaaagatccaaacgaaaagagagatcacatggtcttgttagaatttgttactgctgctggtattacccatggtatggatgaattgtacaaaggatccGGATCC +mTurquoise2(Scerevisae),pOpen_v3,dna,AatggtttctaaaggtgaagaattattcactggtgttgtcccaattttggttgaattagatggtgatgttaatggtcacaaattttctgtctccggtgaaggtgaaggtgatgctacttacggtaaattgaccttaaaatttatttgtactactggtaaattgccagttccatggccaaccttagtcactactttatcttggggtgttcaatgttttgcaagatacccagatcatatgaaacaacatgactttttcaagtctgccatgccagaaggttatgttcaagaaagaactatttttttcaaagatgacggtaactacaagaccagagctgaagtcaagtttgaaggtgataccttagttaatagaatcgaattaaaaggtattgattttaaagaagatggtaacattttaggtcacaaattggaatacaattatttctctgacaatgtttacatcactgctgacaaacaaaagaatggtatcaaagctaacttcaaaattagacacaacattgaagatggtggtgttcaattagctgaccattatcaacaaaatactccaattggtgatggtccagtcttgttaccagacaaccattacttatccactcaatctaagttatccaaagatccaaacgaaaagagggaccacatggtcttgttagaatttgttactgctgctggtattaccttgggtatggatgaattgtacaaaGGATCC +tENO1_terminator,pOpen_v3,dna,ATCCTAAagcttttgattaagccttctagtccaaaaaacacgtttttttgtcatttatttcattttcttagaatagtttagtttattcattttatagtcacgaatgttttatgattctatatagggttgcaaacaagcatttttcattttatgttaaaacaatttcaggtttaccttttattctgcttgtggtgacgcgtgtatccgcccgctcttttggtcacccatgtatCGCT +tSSA1_terminator,pOpen_v3,dna,ATCCTAAgccaattggtgcggcaattgataataacgaaaatgtcttttaatgatctgggtataatgaggaattttccgaacgtttttactttatatatatatatacatgtaacatatattctatacgctatagagaaaggaaatttttcaattaaaaaaaaaatagagaaagagtttcacttcttgattatcgctaacactaatggttgaagtactgctactttaattttatCGCT +tADH1_terminator,pOpen_v3,dna,ATCCTAAgcgaatttcttatgatttatgatttttattattaaataagttataaaaaaaataagtgtatacaaattttaaagtgactcttaggttttaaaacgaaaattcttattcttgagtaactctttcctgtaggtcaggttgctttctcaggtatagcatgaggtcgctcttattgaccacacctctaccggcatgccgagcaaatgcctgcaaatcgctccccatttcCGCT +tPGK1_terminator,pOpen_v3,dna,ATCCTAAattgaattgaattgaaatcgatagatcaatttttttcttttctctttccccatcctttacgctaaaataatagtttattttattttttgaatattttttatttatatacgtatatatagactattatttatcttttaatgattattaagatttttattaaaaaaaaattcgctcctcttttaatgcctttatgcagtttttttttcccattcgatatttctatgtCGCT +tENO2_terminator,pOpen_v3,dna,ATCCTAAagtgcttttaactaagaattattagtcttttctgcttattttttcatcatagtttagaacactttatattaacgaatagtttatgaatctatttaggtttaaaaattgatacagttttataagttactttttcaaagactcgtgctgtctattgcataatgcactggaaggggaaaaaaaaggtgcacacgcgtggctttttcttgaatttgcagtttgaaaaatCGCT +tTDH1_terminator,pOpen_v3,dna,ATCCTAAataaagcaatcttgatgaggataatgatttttttttgaatatacataaatactaccgtttttctgctagattttgtgatgacgtaaataagtacatattactttttaagccaagacaagattaagcattaactttacccttttctttctaagtttcaatattagttatcactgtttaaaagttatggcgagaacgtcggcggttaaaatatattaccctgaacgCGCT +tENO1_terminator_ctag,pOpen_v3,dna,ATGTagcttttgattaagccttctagtccaaaaaacacgtttttttgtcatttatttcattttcttagaatagtttagtttattcattttatagtcacgaatgttttatgattctatatagggttgcaaacaagcatttttcattttatgttaaaacaatttcaggtttaccttttattctgcttgtggtgacgcgtgtatccgcccgctcttttggtcacccatgtatCGCT +tSSA1_terminator_ctag,pOpen_v3,dna,ATGTgccaattggtgcggcaattgataataacgaaaatgtcttttaatgatctgggtataatgaggaattttccgaacgtttttactttatatatatatatacatgtaacatatattctatacgctatagagaaaggaaatttttcaattaaaaaaaaaatagagaaagagtttcacttcttgattatcgctaacactaatggttgaagtactgctactttaattttatCGCT +tADH1_terminator_ctag,pOpen_v3,dna,ATGTgcgaatttcttatgatttatgatttttattattaaataagttataaaaaaaataagtgtatacaaattttaaagtgactcttaggttttaaaacgaaaattcttattcttgagtaactctttcctgtaggtcaggttgctttctcaggtatagcatgaggtcgctcttattgaccacacctctaccggcatgccgagcaaatgcctgcaaatcgctccccatttcCGCT +tPGK1_terminator_ctag,pOpen_v3,dna,ATGTattgaattgaattgaaatcgatagatcaatttttttcttttctctttccccatcctttacgctaaaataatagtttattttattttttgaatattttttatttatatacgtatatatagactattatttatcttttaatgattattaagatttttattaaaaaaaaattcgctcctcttttaatgcctttatgcagtttttttttcccattcgatatttctatgtCGCT +tENO2_terminator_ctag,pOpen_v3,dna,ATGTagtgcttttaactaagaattattagtcttttctgcttattttttcatcatagtttagaacactttatattaacgaatagtttatgaatctatttaggtttaaaaattgatacagttttataagttactttttcaaagactcgtgctgtctattgcataatgcactggaaggggaaaaaaaaggtgcacacgcgtggctttttcttgaatttgcagtttgaaaaatCGCT +tTDH1_terminator_ctag,pOpen_v3,dna,ATGTataaagcaatcttgatgaggataatgatttttttttgaatatacataaatactaccgtttttctgctagattttgtgatgacgtaaataagtacatattactttttaagccaagacaagattaagcattaactttacccttttctttctaagtttcaatattagttatcactgtttaaaagttatggcgagaacgtcggcggttaaaatatattaccctgaacgCGCT +URA3,pOpen_v3,dna,AAGCcggtttccttgaaatttttttgattcggtaatctccgaacagaaggaagaacgaaggaaggagcacagacttagattggtatatatacgcatatgtagtgttgaagaaacatgaaattgcccagtattcttaacccaactgcacagaacaaaaacgtgcaggaaacgaagataaatcatgtcgaaagctacatataaggaacgtgctgctactcatcctagtcctgttgctgccaagctatttaatatcatgcacgaaaagcaaacaaacttgtgtgcttcattggatgttcgtaccaccaaggaattactggagttagttgaagcattaggtcccaaaatttgtttactaaaaacacatgtggatatcttgactgatttttccatggagggcacagttaagccgctaaaggcattatccgccaagtacaattttttactcttcgaggacagaaaatttgctgacattggtaatacagtcaaattgcagtactctgcgggtgtatacagaatagcagaatgggcagacattacgaatgcacacggtgtggtgggcccaggtattgttagcggtttgaagcaggcggcagaagaagtaacaaaggaacctagaggccttttgatgttagcagaattgtcatgcaagggctccctatctactggagaatatactaagggtactgttgacattgcgaagagcgacaaagattttgttatcggctttattgctcaaagagacatgggtggaagagatgaaggttacgattggttgattatgacacccggtgtgggtttagatgacaagggagatgcattgggtcaacagtatagaaccgtggatgatgtggtttctacaggatctgacattattattgttggaagaggactatttgcaaagggaagggatgctaaggtagagggtgaacgttacagaaaagcaggctgggaagcatatttgagaagatgcggccagcaaaactaaaaaactgtattataagtaaatgcatgtatactaaactcacaaattagagcttcaatttaattatatcagttattacccATAG +LEU2,pOpen_v3,dna,AAGCtcgaggagaacttctagtatatctacatacctaatattattgccttattaaaaatggaatcccaacaattacatcaaaatccacattctcttcaaaatcaattgtcctgtacttccttgttcatgtgtgttcaaaaacgttatatttataggataattatactctatttctcaacaagtaattggttgtttggccgagcggtctaaggcgcctgattcaagaaatatcttgaccgcagttaactgtgggaatactcaggtatcgtaagatgcaagagttcgaatctcttagcaaccattatttttttcctcaacataacgagaacacacaggggcgctatcgcacagaatcaaattcgatgactggaaattttttgttaatttcagaggtcgcctgacgcatatacctttttcaactgaaaaattgggagaaaaaggaaaggtgagagcgccggaaccggcttttcatatagaatagagaagcgttcatgactaaatgcttgcatcacaatacttgaagttgacaatattatttaaggacctattgttttttccaataggtggttagcaatcgtcttactttctaacttttcttaccttttacatttcagcaatatatatatatatatttcaaggatataccattctaatgtctgcccctaagaagatcgtcgttttgccaggtgaccacgttggtcaagaaatcacagccgaagccattaaggttcttaaagctatttctgatgttcgttccaatgtcaagttcgatttcgaaaatcatttaattggtggtgctgctatcgatgctacaggtgttccacttccagatgaggcgctggaagcctccaagaaggctgatgccgttttgttaggtgctgtgggtggtcctaaatggggaaccggtagtgttagacctgaacaaggtttactaaaaatccgtaaagaacttcaattgtacgccaacttaagaccatgtaactttgcatccgactctcttttagacttatctccaatcaagccacaatttgctaaaggtactgacttcgttgttgtcagagaattagtgggaggtatttactttggtaagagaaaggaggacgatggtgatggtgtcgcttgggatagtgaacaatacaccgttccagaagtgcaaagaatcacaagaatggccgctttcatggccctacaacatgagccaccattgcctatttggtccttggataaagctaatgttttggcctcttcaagattatggagaaaaactgtggaggaaaccatcaagaacgaatttcctacattgaaggttcaacatcaattgattgattctgccgccatgatcctagttaagaacccaacccacctaaatggtattataatcaccagcaacatgtttggtgatatcatctccgatgaagcctccgttatcccaggttccttgggtttgttgccatctgcgtccttggcctctttgccagacaagaacaccgcatttggtttgtacgaaccatgccacggttctgctccagatttgccaaagaataaggtcaaccctatcgccactatcttgtctgctgcaatgatgttgaaattgtcattgaacttgcctgaagaaggtaaggccattgaagatgcagttaaaaaggttttggatgcaggtatcagaactggtgatttaggtggttccaacagtaccaccgaagtcggtgatgctgtcgccgaagaagttaagaaaatccttgcttaacctggaggacccttctctttagactattctactcttatgcacgtaaaaaattctaggaaatatgtattaactaggagtaaaataaccggctagtggcattcatatagccgtctgtttacatctacatcacacatttcgagtgtatatctcgcaacgttggcgttaaataggcagATAG +HIS3,pOpen_v3,dna,AAGCaacacagtcctttcccgcaattttctttttctattactcttggcctcctctagtacactctatatttttttatgcctcggtaatgattttcatttttttttttccacctagcggatgactctttttttttcttagcgattggcattatcacataatgaattatacattatataaagtaatgtgatttcttcgaagaatatactaaaaaatgagcaggcaagataaacgaaggcaaagatgacagagcagaaagccctagtaaagcgtattacaaatgaaaccaagattcagattgcgatctctttaaagggtggtcccctagcgatagagcactcgatcttcccagaaaaagaggcagaagcagtagcagaacaggccacacaatcgcaagtgattaacgtccacacaggtatagggtttctggaccatatgatacatgctctggccaagcattccggctggtcgctaatcgttgagtgcattggtgacttacacatagacgaccatcacaccactgaggactgcgggattgctctcggtcaagcttttaaagaggccctaggggccgtgcgtggagtaaaaaggtttggatcaggatttgcgcctttggatgaggcactttccagagcggtggttgatctttcgaacaggccgtacgcagttgtcgaacttggtttgcaaagggagaaagtaggtgatctctcttgcgagatgatcccgcattttcttgaaagctttgcagaggctagcagaattaccctccacgttgattgtctgcgaggcaagaatgatcatcaccgtagtgagagtgcgttcaaggctcttgcggttgccataagagaagccacctcgcccaatggtaccaacgatgttccctccaccaaaggtgttcttatgtagtgacaccgattatttaaagctgctgcatacgatatatatacatgtgtatatatgtatacctatgaatgtcagtaagtatgtatacgaacagtatgatactgaagatgacaaggtaatgcatcattctatacgtgtcattctgaacgaggcgcgctttccttttttctttttgctttttctttttttttctcttgaactcgacggatcataATAG +KanamycinR,pOpen_v3,dna,AAGCagcttgcctcgtccccgccgggtcacccggccagcgacatggaggcccagaataccctccttgacagtcttgacgtgcgcagctcaggggcatgatgtgactgtcgcccgtacatttagcccatacatccccatgtataatcatttgcatccatacattttgatggccgcacggcgcgaagcaaaaattacggctcctcgctccagacctgcgagcagggaaacgctcccctcacagacgcgttgaattgtccccacgccgcgcccctgtagagaaatataaaaggttaggatttgccactgaggttcttctttcatatacttccttttaaaatcttgctaggatacagttctcacatcacatccgaacataaacaaaaatgggtaaggaaaagactcacgtttcgaggccgcgattaaattccaacatggatgctgatttatatgggtataaatgggctcgcgataatgtcgggcaatcaggtgcgacaatctatcgattgtatgggaagcccgatgcgccagagttgtttctgaaacatggcaaaggtagcgttgccaatgatgttacagatgagatggtcagactaaactggctgacggaatttatgcctcttccgaccatcaagcattttatccgtactcctgatgatgcatggttactcaccactgcgatccccggcaaaacagcattccaggtattagaagaatatcctgattcaggtgaaaatattgttgatgcgctggcagtgttcctgcgccggttgcattcgattcctgtttgtaattgtccttttaacagcgatcgcgtatttcgtctggctcaggcgcaatcacgaatgaataacggtttggttgatgcgagtgattttgatgacgagcgtaatggctggcctgttgaacaagtctggaaagaaatgcataagcttttgccattctcaccggattcagtcgtcactcatggtgatttctcacttgataaccttatttttgacgaggggaaattaataggttgtattgatgttggacgagtcggaatcgcagaccgataccaggatcttgccatcctatggaactgcctcggtgagttttctccttcattacagaaacggctttttcaaaaatatggtattgataatcctgatatgaataaattgcagtttcatttgatgctcgatgagtttttctaaagtaactgacaataaaaagattcttgttttcaagaacttgtcatttgtatagtttttttatattgtagttgttctattttaatcaaatgttagcgtgatttatattttttttcgcctcgacatcatctgcccagatgcgaagttaagtgcgcagaaagtaatatcatgcgtcaatcgtatgtgaatgctggtcgctatactgATAG +NourseothricinR,pOpen_v3,dna,AAGCagcttgcctcgtccccgccgggtcacccggccagcgacatggaggcccagaataccctccttgacagtcttgacgtgcgcagctcaggggcatgatgtgactgtcgcccgtacatttagcccatacatccccatgtataatcatttgcatccatacattttgatggccgcacggcgcgaagcaaaaattacggctcctcgctccagacctgcgagcagggaaacgctcccctcacagacgcgttgaattgtccccacgccgcgcccctgtagagaaatataaaaggttaggatttgccactgaggttcttctttcatatacttccttttaaaatcttgctaggatacagttctcacatcacatccgaacataaacaaaaatgggtactaccttagatgatacagcctacagatacagaacatcagtccctggtgatgctgaagcaattgaggctttagacggttcattcaccaccgacaccgtctttagagtaaccgccaccggtgatggatttaccttaagagaagtcccagtcgaccctccattaactaaagtctttccagatgatgaatctgatgacgaaagcgacgacggagaagatggtgacccagattcaagaactttcgtagcatacggtgatgacggtgatttggctggttttgtagtcgtttcttattcaggttggaatagaaggttgaccgttgaagatatagaagtcgccccagagcatagaggtcatggtgtaggaagagctttgatgggtttggctacagaatttgcaagagagagaggagccggtcatttatggttagaagttactaatgttaacgcccctgctatccatgcttatagaagaatgggtttcacattatgtggtttagatactgctttatatgatggaacagcatctgacggtgaacaggccttgtatatgtctatgccttgcccttaaagtaactgacaataaaaagattcttgttttcaagaacttgtcatttgtatagtttttttatattgtagttgttctattttaatcaaatgttagcgtgatttatattttttttcgcctcgacatcatctgcccagatgcgaagttaagtgcgcagaaagtaatatcatgcgtcaatcgtatgtgaatgctggtcgctatactgATAG +HygromycinR,pOpen_v3,dna,AAGCagcttgcctcgtccccgccgggtcacccggccagcgacatggaggcccagaataccctccttgacagtcttgacgtgcgcagctcaggggcatgatgtgactgtcgcccgtacatttagcccatacatccccatgtataatcatttgcatccatacattttgatggccgcacggcgcgaagcaaaaattacggctcctcgctccagacctgcgagcagggaaacgctcccctcacagacgcgttgaattgtccccacgccgcgcccctgtagagaaatataaaaggttaggatttgccactgaggttcttctttcatatacttccttttaaaatcttgctaggatacagttctcacatcacatccgaacataaacaaaaatgggtaaaaagcctgaactcaccgcgacgtctgtcgagaagtttctgatcgaaaagttcgacagcgtgtccgacctgatgcagctctcggagggcgaagaatctcgtgctttcagcttcgatgtaggagggcgtggatatgtcctgcgggtaaatagctgcgccgatggtttctacaaagatcgttatgtttatcggcactttgcatcggccgcgctcccgattccggaagtgcttgacattggggaatttagcgagagcctgacctattgcatctcccgccgtgcacagggtgtcacgttgcaagacctgcctgaaaccgaactgcccgctgttctgcaaccggtcgcggaggccatggatgcgatcgctgcggccgatcttagccagacgagcgggttcggcccattcggaccgcaaggaatcggtcaatacactacatggcgtgatttcatatgcgcgattgctgatccccatgtgtatcactggcaaactgtgatggacgacaccgtcagtgcgtccgtcgcgcaggctctcgatgagctgatgctttgggccgaggactgccccgaagtccggcacctcgtgcacgcggatttcggctccaacaatgtcctgacggacaatggccgcataacagcggtcattgactggagcgaggcgatgttcggggattcccaatacgaggtcgccaacatcttcttctggaggccgtggttggcttgtatggagcagcagacgcgctacttcgagcggaggcatccggagcttgcaggatcgccgcggctccgggcgtatatgctccgcattggtcttgaccaactctatcagagcttggttgacggcaatttcgatgatgcagcttgggcgcagggtcgatgcgacgcaatcgtccgatccggagccgggactgtcgggcgtacacaaatcgcccgcagaagcgcggccgtctggaccgatggctgtgtagaagtactcgccgatagtggaaaccgacgccccagcactcgtccgagggcaaaggaataaagtaactgacaataaaaagattcttgttttcaagaacttgtcatttgtatagtttttttatattgtagttgttctattttaatcaaatgttagcgtgatttatattttttttcgcctcgacatcatctgcccagatgcgaagttaagtgcgcagaaagtaatatcatgcgtcaatcgtatgtgaatgctggtcgctatactgATAG +ZeocinR,pOpen_v3,dna,AAGCagcttgcctcgtccccgccgggtcacccggccagcgacatggaggcccagaataccctccttgacagtcttgacgtgcgcagctcaggggcatgatgtgactgtcgcccgtacatttagcccatacatccccatgtataatcatttgcatccatacattttgatggccgcacggcgcgaagcaaaaattacggctcctcgctccagacctgcgagcagggaaacgctcccctcacagacgcgttgaattgtccccacgccgcgcccctgtagagaaatataaaaggttaggatttgccactgaggttcttctttcatatacttccttttaaaatcttgctaggatacagttctcacatcacatccgaacataaacaaaaatggctaaattaacatctgccgttcctgttttaacagctagggatgttgcaggtgctgtagagttttggacagataggttaggattctcaagagactttgttgaggacgattttgctggtgttgtcagggatgacgttactttatttatctcagcagtccaagatcaagttgtccctgataatacattggcttgggtctgggtcaggggtttagatgaattatatgctgaatggtcagaagttgtatctacaaacttcagagatgcttctggtccagctatgaccgagattggtgaacagccatggggtagagaatttgctttgagagatccagctggaaattgtgttcattttgttgctgaagaacaagattaaagtaactgacaataaaaagattcttgttttcaagaacttgtcatttgtatagtttttttatattgtagttgttctattttaatcaaatgttagcgtgatttatattttttttcgcctcgacatcatctgcccagatgcgaagttaagtgcgcagaaagtaatatcatgcgtcaatcgtatgtgaatgctggtcgctatactgATAG +2micron,pOpen_v3,dna,GTCTaacgaagcatctgtgcttcattttgtagaacaaaaatgcaacgcgagagcgctaatttttcaaacaaagaatctgagctgcatttttacagaacagaaatgcaacgcgaaagcgctattttaccaacgaagaatctgtgcttcatttttgtaaaacaaaaatgcaacgcgagagcgctaatttttcaaacaaagaatctgagctgcatttttacagaacagaaatgcaacgcgagagcgctattttaccaacaaagaatctatacttcttttttgttctacaaaaatgcatcccgagagcgctatttttctaacaaagcatcttagattactttttttctcctttgtgcgctctataatgcagtctcttgataactttttgcactgtaggtccgttaaggttagaagaaggctactttggtgtctattttctcttccataaaaaaagcctgactccacttcccgcgtttactgattactagcgaagctgcgggtgcattttttcaagataaaggcatccccgattatattctataccgatgtggattgcgcatactttgtgaacagaaagtgatagcgttgatgattcttcattggtcagaaaattatgaacggtttcttctattttgtctctatatactacgtataggaaatgtttacattttcgtattgttttcgattcactctatgaatagttcttactacaatttttttgtctaaagagtaatactagagataaacataaaaaatgtagaggtcgagtttagatgcaagttcaaggagcgaaaggtggatgggtaggttatatagggatatagcacagagatatatagcaaagagatacttttgagcaatgtttgtggaagcggtattcgcaatattttagtagctcgttacagtccggtgcgtttttggttttttgaaagtgcgtcatcagagcgcttttggttttcaaaagcgctctgaagttcctatactttctagctagagaataggaacttcAAGC +CEN_ARS,pOpen_v3,dna,GTCTatcacgtgctataaaaataattataatttaaattttttaatataaatatataaattaaaaatagaaagtaaaaaaagaaattaaagaaaaaatagtttttgttttccgaagatgtaaaagactctagggggatcgccaacaaatactaccttttatcttgctcttcctgctctcaggtattaatgccgaattgtttcatcttgtctgtgtagaaaaccacacacgaaaatcctgtgattttacattttacttatcgttaatcgaatgtatatctatttaatctgcttttcttgtctaataaatatatatgtaaagtacgctttttgttgaaattttttaaacctttgtttatttttttttcttcattccgtaactcttctaccttctttatttactttctaaaatccaaatacaaaacataaaaataaataaacacagagtaaattcccaaattattccatcattaaaagatacgaggcgcgtgtaagttacaggcaagcgatccgtcAAGC +BBa_J23119,pOpen_v3,dna,TACAttgacagctagctcagtcctaggtataatgctagcAACT +BBa_J23100,pOpen_v3,dna,TACAttgacggctagctcagtcctaggtacagtgctagcAACT +BBa_J23101,pOpen_v3,dna,TACAtttacagctagctcagtcctaggtattatgctagcAACT +BBa_J23102,pOpen_v3,dna,TACAttgacagctagctcagtcctaggtactgtgctagcAACT +BBa_J23103,pOpen_v3,dna,TACActgatagctagctcagtcctagggattatgctagcAACT +BBa_J23104,pOpen_v3,dna,TACAttgacagctagctcagtcctaggtattgtgctagcAACT +BBa_J23105,pOpen_v3,dna,TACAtttacggctagctcagtcctaggtactatgctagcAACT +BBa_J23106,pOpen_v3,dna,TACAtttacggctagctcagtcctaggtatagtgctagcAACT +BBa_J23107,pOpen_v3,dna,TACAtttacggctagctcagccctaggtattatgctagcAACT +BBa_J23108,pOpen_v3,dna,TACActgacagctagctcagtcctaggtataatgctagcAACT +BBa_J23109,pOpen_v3,dna,TACAtttacagctagctcagtcctagggactgtgctagcAACT +BBa_J23110,pOpen_v3,dna,TACAtttacggctagctcagtcctaggtacaatgctagcAACT +BBa_J23111,pOpen_v3,dna,TACAttgacggctagctcagtcctaggtatagtgctagcAACT +BBa_J23112,pOpen_v3,dna,TACActgatagctagctcagtcctagggattatgctagcAACT +BBa_J23113,pOpen_v3,dna,TACActgatggctagctcagtcctagggattatgctagcAACT +BBa_J23114,pOpen_v3,dna,TACAtttatggctagctcagtcctaggtacaatgctagcAACT +BBa_J23115,pOpen_v3,dna,TACAtttatagctagctcagcccttggtacaatgctagcAACT +BBa_J23116,pOpen_v3,dna,TACAttgacagctagctcagtcctagggactatgctagcAACT +BBa_J23117,pOpen_v3,dna,TACAttgacagctagctcagtcctagggattgtgctagcAACT +BBa_J23118,pOpen_v3,dna,TACAttgacggctagctcagtcctaggtattgtgctagcAACT +Vnat_dummy_promoter,pOpen_v3,dna,TACAcccctggcgccccttAACT +Ptrc_promoter,pOpen_v3,dna,TACAgtctagggcggcggatttgtcctactcaggagagcgttcaccgacaaacaacagataaaacgaaaggcccagtctttcgactgagcctttcgttttatttgatgcagcgggtcgactcccctaggtcactgcccgctttccagtcgggaaacctgtcgtgccagctgcattaatgaatcggccaacgcgcggggagaggcggtttgcgtattgggcgccagggtggtttttcttttcaccagtgacacgggcaacagctgattgcccttcaccgcctggccctgagagagttgcagcaagcggtccacgctggtttgccccagcaggcgaaaatcctgtttgatggtggttaacggcgggatataacatgagctatcttcggtatcgtcgtatcccactaccgagatatccgcaccaacgcgcagcccggactcggtaatggcgcgcattgcgcccagcgccatctgatcgttggcaaccagcatcgcagtgggaacgatgccctcattcagcatttgcatggtttgttgaaaaccggacatggcactccagtcgccttcccgttccgctatcggctgaatttgattgcgagtgagatatttatgccagccagccagacgcagacgcgccgagacagaacttaatgggcccgctaacagcgcgatttgctggtgacccaatgcgaccagatgctccacgcccagtcgcgtaccatcttcatgggagaaaataatactgttgatgggtgtctggtcagagacatcaagaaataacgccggaacattagtgcaggcagcttccacagcaatggcatcctggtcatccagcggatagttaatgatcagcccactgacgcgttgcgcgagaagattgtgcaccgccgctttacaggcttcgacgccgcttcgttctaccatcgacaccaccacgctggcacccagttgatcggcgcgagatttaatcgccgcgacaatttgcgacggcgcgtgcagggccagactggaggtggcaacgccaatcagcaacgactgtttgcccgccagttgttgtgccacgcggttgggaatgtaattcagctccgccatcgccgcttccactttttcccgcgttttcgcagaaacgtggctggcctggttcaccacgcgggaaacggtctgataagagacaccggcatactctgcgacatcgtataacgttactggtttcacattcaccaccctgaattgactctcttccgggcgctatcatgccataccgcgaaaggttttgcgccaaagctttccctcgacaattcgataaatgtgagcggataacattgacattggtgagcggataacaagatactgagcacatcagcaggacgcactgaccAACT +Ptet_promoter,pOpen_v3,dna,TACAttttgttatcaataaaaaaggccccccgttagggaggcttattgttcgtcccatcacggaaaaaggttatgctgcttttaagacccactttcacatttaagttgtttttctaatccgcatatgatcaattcaaggccgaataagaaggctggctctgcaccttggtgatcaaataattcgatagcttgtcgtaataatggcggcatactatcagtagtaggtgtttccctttcttctttagcgacttgatgctcttgatcttccaatacgcaacctaaagtaaaatgccccacagcgctgagtgcatataatgcattctctagtgaaaaaccttgttggcataaaaaggctaattgattttcgagagtttcatactgtttttctgtaggccgtgtacctaaatgtacttttgctccatcgcgatgacttagtaaagcacatctaaaacttttagcgttattacgtaaaaaatcttgccagctttccccttctaaagggcaaaagtgagtatggtgcctatctaacatctcaatggctaaggcgtcgagcaaagcccgcttattttttacatgccaatacaatgtaggctgctctacacctagcctctgggcgagtttacgggttgttaaaccttcgattccgacctcattaagcagctctaatgcgctgctaatcactctacttttatctaatcgagacatcattaattcctaatttttgttgacactctatcattgatagagttattttaccactccctatcagtgatagagaaaagtgaaAACT +Para(Vn)_promoter,pOpen_v3,dna,TACAtgttatccatccactggtagaggtgagtgttcgctatacatatttgtggatttagggacatttgttagtgacaaaaatcacagcggaaaaatgtagcgaatttgtccattcatttagccagtgtggctatgacacagatctcaattatgcgaccaatgatccaaattctcagtaagcaacccaataccagcctaatgcaaacgttgaattcgctggtttcttgcttttccgacctgacagaagaaggtgttaaaaagaacacaaaaaatcgtccatgacgtttttgtccataaagttagcagacctctttatgggataacatccctcctctagctataacaacaagtagatttagtttgctgaccaaaaAACT +Prham(Vn)_promoter,pOpen_v3,dna,TACAgacacactctaataaccaagccccgcaattcgcggggcttattatttttagccagccaaatgttacgccctccccgttatttcaaacagtaaatagcttgaataaataagaaaaaccacaccttttacacgcctactccacttcacttaaacccaggttttatctggcctcacgcacgagttgtcaaaagtttgaattaccgcaagagcttcttgagaaaaacgcatcgaatacgttttttcagggggatttttgaagttatttagtgcggaaaaacggtgtaatacctctaaagaacaagaggtgttaatcAACT +Vnat_dummy_rbs,pOpen_v3,dna,AACTagagtgtcaggatacccgataatcAATG +B0029_rbs,pOpen_v3,dna,AACTagagttcacacaggaaacctaatcAATG +B0030_rbs,pOpen_v3,dna,AACTagagattaaagaggagaaataatcAATG +B0031_rbs,pOpen_v3,dna,AACTagagtcacacaggaaacctaatcAATG +B0032_rbs,pOpen_v3,dna,AACTagagtcacacaggaaagtaatcAATG +B0033_rbs,pOpen_v3,dna,AACTagagtcacacaggactaatcAATG +B0034_rbs,pOpen_v3,dna,AACTagagaaagaggagaaataatcAATG +B0035_rbs,pOpen_v3,dna,AACTagagattaaagaggagaataatcAATG +B0064_rbs,pOpen_v3,dna,AACTagagaaagaggggaaataatcAATG +lux_operon(Vn),pOpen_v3,dna,AatgactaaaaaaatttcattcattattaacggccaggttgaaatctttcccgaaagtgatgatttagtgcaatccattaattttggtgataatagtgtttacctgccaatattgaatgactctcatgtaaaaaacattattgattgtaatggaaataacgaattacggttgcataacattgtcaattttctctatacggtagggcaaagatggaaaaatgaagaatactcaagacgcaggacatacattcgtgacttaaaaaaatatatgggatattcagaagaaatggctaagctagaggccaattggatatctatgattttatgttctaaaggcggcctttatgatgttgtagaaaatgaacttggttctcgccatatcatggatgaatggctacctcaggatgaaagttatgttcgggcttttccgaaaggtaaatctgtacatctgttggcaggtaatgttccattatctgggatcatgtctatattacgcgcaattttaactaagaatcagtgtattataaaaacatcgtcaaccgatccttttaccgctaatgcattagcgttaagttttattgatgtagaccctaatcatccgataacgcgctctttatctgttatatattggccccaccaaggtgatacatcactcgcaaaagaaattatgcgacatgcggatgttattgtcgcttggggagggccagatgcgattaattgggcggtagagcacgcgccatcttatgctgatgtgattaaatttggttctaaaaagagtctttgcattatcgataatcctgttgatttgacgtccgcagcgacaggtgcggctcatgatgtttgtttttacgatcagcgagcttgtttttctgcccaaaacatatattacatgggaaatcattatgaggaatttaagttagcgttgatagaaaaacttaatctatatgcgcatatattaccgaatgccaaaaaagattttgatgaaaaggcggcctattctttagttcaaaaagaaagcctgtttgctggattaaaagtagaggtggatattcatcaacgttggatgattattgagtcaaatgcaggtgtggaatttaatcaaccacttggcagatgtgtgtaccttcatcacgtcgataatattgagcaaatattgccttatgttcaaaaaaataagacgcaaaccatatctatttttccttgggagtcatcatttaaatatcgagatgcgttagcattaaaaggtgcggaaaggattgtagaagcaggaatgaataacatatttcgagttggtggatctcatgacggaatgaggccgttgcaacgattagtgacatatatttctcatgaaaggccatctaactatacggctaaggatgttgcggttgaaatagaacagactcgattcctggaagaagataagttccttgtatttgtcccataataggtaaaaagtatggaaaatgaatcaaaatataaaaccatcgaccacgttatttgtgttgaaggaaataaaaaaattcatgtttgggaaacgctgccagaagaaaacagcccaaagagaaagaatgccattattattgcgtctggttttgcccgcaggatggatcattttgctggtctggcggaatatttatcgcggaatggatttcatgtgatccgctatgattcgcttcaccatgttggcttgagttcagggacaattgatgaatttacaatgtctataggaaagcagagcttgttagcagtggttgattggttaactacacgaaaaataaataacttcggtatgttggcttccagcttatctgcgcggatagcttatgcaagcctatctgaaatcaatgcttcgtttttaatcaccgcagtcggtgttgttaacttaagatattctcttgaaagagctttagggtttgattatctcagtctacccattaatgaattgccggataatctggattttgaaggccataaattgggtgctgaagtctttgcgagagattgtcttgattttggttgggaagatttagcttctacaattaataacatgatgtatcttgatataccgtttattgcttttactgcaaataacgataattgggtcaagcaagatgaagttatcacattgttatcaaatattcgtagtaatcgatgcaagatatattctttgttaggaagttcgcatgacttgagtgaaaatttagtggtcctgcgcaatttttatcaatcggttacgaaagccgctatcgcgatggataatgatcatctggatattgatgttgatattactgaaccgtcatttgaacatttaactattgcgacagtcaatgaacgccgaatgagaattgagattgaaaatcaagcaatttctctgtcttaaaatctattgagatattctatcactcaaatagcaatataaggactctctatgaaatttggaaactttttgcttacataccaacctccccaattttctcaaacagaggtaatgaaacgtttggttaaattaggtcgcatctctgaggagtgtggttttgataccgtatggttactggagcatcatttcacggagtttggtttgcttggtaacccttatgtcgctgctgcatatttacttggcgcgactaaaaaattgaatgtaggaactgccgctattgttcttcccacagcccatccagtacgccaacttgaagatgtgaatttattggatcaaatgtcaaaaggacgatttcggtttggtatttgccgagggctttacaacaaggactttcgcgtattcggcacagatatgaataacagtcgcgccttagcggaatgctggtacgggctgataaagaatggcatgacagagggatatatggaagctgataatgaacatatcaagttccataaggtaaaagtaaaccccgcggcgtatagcagaggtggcgcaccggtttatgtggtggctgaatcagcttcgacgactgagtgggctgctcaatttggcctaccgatgatattaagttggattataaatactaacgaaaagaaagcacaacttgagctttataatgaagtggctcaagaatatgggcacgatattcataatatcgaccattgcttatcatatataacatctgtagatcatgactcaattaaagcgaaagagatttgccggaaatttctggggcattggtatgattcttatgtgaatgctacgactatttttgatgattcagaccaaacaagaggttatgatttcaataaagggcagtggcgtgactttgtattaaaaggacataaagatactaatcgccgtattgattacagttacgaaatcaatcccgtgggaacgccgcaggaatgtattgacataattcaaaaagacattgatgctacaggaatatcaaatatttgttgtggatttgaagctaatggaacagtagacgaaattattgcttccatgaagctcttccagtctgatgtcatgccatttcttaaagaaaaacaacgttcgctattatattagctaaggagaaagaaatgaaatttggattgttcttccttaacttcatcaattcaacaactgttcaagaacaaagtatagttcgtatgcaggaaataacggagtatgttgataagttgaattttgaacagattttagtgtatgaaaatcatttttcagataatggtgttgtcggcgctcctctgactgtttctggttttctgctcggtttaacagagaaaattaaaattggttcattaaatcacatcattacaactcatcatcctgtccgcatagcggaggaagcgtgcttattggatcagttaagtgaagggagatttattttagggtttagtgattgcgaaaaaaaagatgaaatgcatttttttaatcgcccggttgaatatcaacagcaactatttgaagagtgttatgaaatcattaacgatgctttaacaacaggctattgtaatccagataacgatttttatagcttccctaaaatatctgtaaatccccatgcttatacgccaggcggacctcggaaatatgtaacagcaaccagtcatcatattgttgagtgggcggccaaaaaaggtattcctctcatctttaagtgggatgattctaatgatgttagatatgaatatgctgaaagatataaagccgttgcggataaatatgacgttgacctatcagagatagaccatcagttaatgatattagttaactataacgaagatagtaataaagctaaacaagagactcgtgcatttattagtgattatgttcttgaaatgcaccctaatgaaaatttcgaaaataaacttgaagaaataattgcagaaaacgctgtcggaaattatacggagtgtataactgcggctaagttggcaattgaaaagtgtggtgcgaaaagtgtattgctgtcctttgaaccaatgaatgatttgatgagccaaaaaaatgtaatcaatattgttgatgataatattaagaagtaccacatggaatatacctaatagatttcgagttgcagcgaggcggcaagtgaacgaatccccaggagcatagataactatgtgactggggtgagtgaaagcagccaacaaagcagcagcttgaaagatgaagggtataaaagagtatgacagcagtgctgccatactttctaatattatcttgaggagtaaaacaggtatgacttcatatgttgataaacaagaaattacagcaagctcagaaattgatgatttgattttttcgagcgatccattagtgtggtcttacgacgagcaggaaaaaatcagaaagaaacttgtgcttgatgcatttcgtaatcattataaacattgtcgagaatatcgtcactactgtcaggcacacaaagtagatgacaatattacggaaattgatgacatacctgtattcccaacatcggtttttaagtttactcgcttattaacttctcaggaaaacgagattgaaagttggtttaccagtagcggcacgaatggtttaaaaagtcaggtggcgcgtgacagattaagtattgagagactcttaggctctgtgagttatggcatgaaatatgttggtagttggtttgatcatcaaatagaattagtcaatttgggaccagatagatttaatgctcataatatttggtttaaatatgttatgagtttggtggaattgttatatcctacgacatttaccgtaacagaagaacgaatagattttgttaaaacattgaatagtcttgaacgaataaaaaatcaagggaaagatctttgtcttattggttcgccatactttatttatttactctgccattatatgaaagataaaaaaatctcattttctggagataaaagcctttatatcataaccggaggcggctggaaaagttacgaaaaagaatctctgaaacgtgatgatttcaatcatcttttatttgatactttcaatctcagtgatattagtcagatccgagatatatttaatcaagttgaactcaacacttgtttctttgaggatgaaatgcagcgtaaacatgttccgccgtgggtatatgcgcgagcgcttgatcctgaaacgttgaaacctgtacctgatggaacgccggggttgatgagttatatggatgcgtcagcaaccagttatccagcatttattgttaccgatgatgtcgggataattagcagagaatatggtaagtatcccggcgtgctcgttgaaattttacgtcgcgtcaatacgaggacgcagaaagggtgtgctttgagcttaaccgaagcgtttgatagtGGATCC +azurite(Vn),pOpen_v3,dna,AatgtctaaaggtgaagaactgtttaccggtgttgttccaattctggttgaactggatggtgatgttaacggtcacaaattttctgtttctggtgaaggcgaaggtgatgcaacctacggtaaactgaccctgaaatttatctgtaccactggtaaactgccagttccatggccaactctggttaccactctgtctcacggtgttcaatgtttttctcgttacccagatcacatgaaacagcacgattttttcaaaagcgcaatgccagaaggttacgttcaagaacgtaccatcttcttcaaagatgacggtaactacaaaacccgtgcggaagtgaaatttgaaggtgataccctggttaaccgtatcgaactgaaaggtatcgacttcaaagaggatggcaacattctgggtcacaaactggaatacaactttaactctcacaacatctacatcatggcggacaaacaaaaaaacggcatcaaagtgaacttcaagattcgccacaacatcgaagatggttctgttcaactggcagatcactaccaacaaaacaccccaattggtgatggtccagttctgctgccagataaccactacctgtctacccaatctgcactgtctaaagatccaaacgaaaaacgtgatcacatggtgctgctggaatttcgtaccgcagcaggtattacccacggtatggatgaactgtacaaaGGATCC +mTurquoise(Vn),pOpen_v3,dna,AatggtttctaaaggtgaagaactgtttaccggtgttgttccaattctggttgaactggatggtgatgttaacggtcacaaattttctgtttctggtgaaggcgaaggtgatgcaacctacggtaaactgaccctgaaatttatctgtaccactggtaaactgccagttccatggccaactctggttaccactctgtcttggggtgttcaatgttttgcacgttacccagatcacatgaaacaacacgattttttcaaaagcgcaatgccagaaggttacgttcaagaacgtaccatcttcttcaaagatgacggtaactacaaaacccgtgcggaagtgaaatttgaaggtgataccctggttaaccgtatcgaactgaaaggtatcgacttcaaagaggatggcaacattctgggtcacaaactggaatacaactactttagcgataacgtgtacatcaccgcagataaacaaaaaaacggtatcaaggcgaacttcaaaatccgtcacaacattgaagatggtggtgttcaactggcagatcactaccaacaaaacactccaattggtgatggtccagttctgctgccagataaccactacctgtctacccaatctaaactgtctaaagacccaaacgaaaaacgtgatcacatggttctgctggaatttgttaccgcagcaggtattaccttaggtatggatgaactgtacaaaGGATCC +sfGFP(Vn),pOpen_v3,dna,AatgcgtaaaggtgaagaactgtttaccggtgttgttccaattctggttgaactggatggtgatgttaacggtcacaaattttctgttcgtggtgaaggcgaaggtgatgcaaccaacggtaaactgaccctgaaatttatctgtaccactggtaaactgccagttccatggccaactctggttaccactctgacctacggtgttcaatgttttgcacgttacccagatcacatgaaacaacacgattttttcaaaagcgcaatgccagaaggttacgttcaagaacgtaccatctcttttaaagatgacggcacctacaaaacccgtgcggaagttaaatttgaaggtgataccctggttaaccgcattgaactgaaaggcatcgattttaaagaagatggtaacatcctgggccacaaactggaatacaactttaactctcacaacgtgtacatcaccgcagacaaacaaaaaaacggtatcaaagcgaacttcaagatccgtcacaacgttgaagatggttctgttcaactggcagatcactaccaacaaaacaccccaattggtgatggtccagttctgctgccagataaccactacctgtctacccaaagcgttctgtctaaagatccaaacgaaaaacgtgatcacatggtgctgctggaatttgttaccgcagcaggtattacccacggtatggatgaactgtacaaagcaGGATCC +mVenus(Vn),pOpen_v3,dna,AatggtttctaaaggtgaagaactgtttaccggtgttgttccaattctggttgaactggatggtgatgttaacggtcacaaattttctgtttctggtgaaggcgaaggtgatgcaacctacggtaaactgaccctgaaactgatttgtaccactggtaaactgccagttccatggccaactctggttaccaccttaggttacggtctgcaatgttttgcacgttacccagatcacatgaaacaacacgattttttcaaaagcgcaatgccagaaggttacgttcaagaacgtaccatcttcttcaaagatgacggtaactacaaaacccgtgcggaagtgaaatttgaaggtgataccctggttaaccgtatcgaactgaaaggtatcgacttcaaagaggatggcaacattctgggtcacaaactggaatacaactacaactctcacaacgtttacatcaccgcagataaacaaaaaaacggcatcaaagcgaacttcaaaatccgtcacaacattgaagatggtggtgttcaactggcagatcactaccaacaaaacaccccaattggtgatggtccagttctgctgccagataaccactacctgtcttaccaatctaaactgagcaaagacccaaacgaaaaacgtgatcacatggttctgctggaatttgttaccgcagcaggtattaccttaggtatggatgaactgtacaaaGGATCC +mScarlet-I(Vn),pOpen_v3,dna,AatggtttctaaaggtgaagcagtgatcaaagaatttatgcgcttcaaagttcacatggaaggttctatgaacggccacgaatttgaaattgaaggtgaaggcgaaggtcgtccatacgaaggtactcaaaccgcaaaactgaaagttaccaaaggtggtccactgccattttcttgggatattctgtctccacaatttatgtacggttctcgtgcatttatcaaacacccagcagatattccagactactacaaacaatcttttccggaaggtttcaaatgggaacgtgttatgaattttgaagatggtggtgcagttacggttacccaagatacctctctggaagatggtactctgatctacaaagttaaactgcgtggtactaactttccaccagatggtccagttatgcagaaaaaaaccatgggttgggaagcatctaccgaacgtctgtacccagaagatggcgttctgaaaggtgatatcaaaatggcactgcgtctgaaagatggcggtcgttacctggcagatttcaaaaccacctacaaagcgaaaaaaccagttcaaatgccaggtgcatacaacgttgatcgtaaactggatattaccagccacaacgaagattacaccgttgttgaacaatacgaacgttctgaaggccgtcactctaccggtggtatggatgaactgtacaaaGGATCC +mCherry(Vn),pOpen_v3,dna,AatggtttctaaaggtgaagaggataacatggcgatcatcaaagaatttatgcgcttcaaagttcacatggaaggttctgttaacggccacgaatttgaaattgaaggtgaaggcgaaggtcgtccatacgaaggtactcaaaccgcaaaactgaaagttaccaaaggtggtccactgccatttgcatgggatattctgtctccacagtttatgtacggtagcaaagcatacgttaaacacccagcagatattccagattacctgaaactgtcttttccggaaggtttcaaatgggaacgtgttatgaattttgaagatggtggtgttgttacggttacccaagattcttctctgcaagatggtgagtttatctacaaagttaaactgcgtggcaccaactttccatctgatggtccagttatgcagaaaaaaaccatgggttgggaagcatcttctgaacgtatgtacccagaagatggcgcactgaaaggtgaaattaaacaacgtctgaaacttaaagatggcggtcactacgatgcagaagttaaaaccacctacaaagcgaaaaaaccagttcaactgccaggtgcatacaacgttaacattaaactggatatcaccagccacaacgaagattacaccattgttgaacaatacgaacgtgcagaaggccgtcactctaccggtggtatggatgaactgtacaaaGGATCC +mKate-2(Vn),pOpen_v3,dna,AatggtttctgaactgattaaagaaaacatgcacatgaaactgtacatggaaggtactgttaacaaccaccacttcaaatgtacctctgaaggtgaaggtaaaccatacgaaggtactcaaaccatgcgtattaaagcagttgaaggtggtccactgccatttgcatttgatattctggcaacctcttttatgtacggcagcaaaacctttatcaaccacactcaaggtatcccggattttttcaaacaaagctttccagaaggtttcacctgggaacgtgttaccacctacgaagatggtggtgttctgaccgcaactcaagatacctctctgcaagatggttgtctgatctacaacgttaaaatccgtggtgttaactttccatctaacggtccagttatgcagaaaaaaaccttaggttgggaagcatctaccgaaactctgtacccagcggatggtggtctggaaggtcgtgcagatatggcactgaaactggttggtggtggtcacctgatttgtaacctgaaaaccacctaccgttctaaaaaaccagcgaaaaatctgaaaatgccaggtgtttactacgttgatcgtcgtctggaacgtatcaaagaagcagataaagaaacctacgtggaacaacacgaagttgcagttgcacgttactgtgatctgccatctaaactgggtcaccgtGGATCC +B0010_terminator,pOpen_v3,dna,ATCCTAAccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctcCGCT +B0015_terminator,pOpen_v3,dna,ATCCTAAccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttataCGCT +B1002_terminator,pOpen_v3,dna,ATCCTAAcgcaaaaaaccccgcttcggcggggttttttcgcCGCT +B1003_terminator,pOpen_v3,dna,ATCCTAAcgccaaaaaccccgcttcggcggggtttttccgcCGCT +B1004_terminator,pOpen_v3,dna,ATCCTAAcgccgaaaaccccgcttcggcggggttttgccgcCGCT +B1005_terminator,pOpen_v3,dna,ATCCTAAcgccgcaaaccccgcttcggcggggtttcgccgcCGCT +B1006_terminator,pOpen_v3,dna,ATCCTAAaaaaaaaaaccccgcccctgacagggcggggttttttttCGCT +B1007_terminator,pOpen_v3,dna,ATCCTAAcgcaaaaaaccccgcccctgacagggcggggttttttcgcCGCT +B1009_terminator,pOpen_v3,dna,ATCCTAAcgccgaaaaccccgcccctgacagggcggggttttgccgcCGCT +B1010_terminator,pOpen_v3,dna,ATCCTAAcgccgcaaaccccgcccctgacagggcggggtttcgccgcCGCT +Vnat_dummy_terminator,pOpen_v3,dna,ATCCTAAactcagttgtagtaacgagcggatagattcccagacccaccttcacgggcggtagcaggacctcaataataggattttcgCGCT +B0010_terminator_ctag,pOpen_v3,dna,ATGTccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctcCGCT +B0015_terminator_ctag,pOpen_v3,dna,ATGTccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttataCGCT +B1002_terminator_ctag,pOpen_v3,dna,ATGTcgcaaaaaaccccgcttcggcggggttttttcgcCGCT +B1003_terminator_ctag,pOpen_v3,dna,ATGTcgccaaaaaccccgcttcggcggggtttttccgcCGCT +B1004_terminator_ctag,pOpen_v3,dna,ATGTcgccgaaaaccccgcttcggcggggttttgccgcCGCT +B1005_terminator_ctag,pOpen_v3,dna,ATGTcgccgcaaaccccgcttcggcggggtttcgccgcCGCT +B1006_terminator_ctag,pOpen_v3,dna,ATGTaaaaaaaaaccccgcccctgacagggcggggttttttttCGCT +B1007_terminator_ctag,pOpen_v3,dna,ATGTcgcaaaaaaccccgcccctgacagggcggggttttttcgcCGCT +B1009_terminator_ctag,pOpen_v3,dna,ATGTcgccgaaaaccccgcccctgacagggcggggttttgccgcCGCT +B1010_terminator_ctag,pOpen_v3,dna,ATGTcgccgcaaaccccgcccctgacagggcggggtttcgccgcCGCT +Vnat_dummy_terminator_ctag,pOpen_v3,dna,ATGTactcagttgtagtaacgagcggatagattcccagacccaccttcacgggcggtagcaggacctcaataataggattttcgCGCT +3xFlag(Vn)_ctag,pOpen_v3,dna,ATCCgattataaggatcatgatggtgattataaggatcatgatatcgactacaaagacgatgacgacaagTAAATGT +6xHis(Vn)_ctag,pOpen_v3,dna,ATCCcaccatcaccaccatcatTAAATGT +Azurite(Vn)_ctag,pOpen_v3,dna,ATCCatgtctaaaggtgaagaactgtttaccggtgttgttccaattctggttgaactggatggtgatgttaacggtcacaaattttctgtttctggtgaaggcgaaggtgatgcaacctacggtaaactgaccctgaaatttatctgtaccactggtaaactgccagttccatggccaactctggttaccactctgtctcacggtgttcaatgtttttctcgttacccagatcacatgaaacagcacgattttttcaaaagcgcaatgccagaaggttacgttcaagaacgtaccatcttcttcaaagatgacggtaactacaaaacccgtgcggaagtgaaatttgaaggtgataccctggttaaccgtatcgaactgaaaggtatcgacttcaaagaggatggcaacattctgggtcacaaactggaatacaactttaactctcacaacatctacatcatggcggacaaacaaaaaaacggcatcaaagtgaacttcaagattcgccacaacatcgaagatggttctgttcaactggcagatcactaccaacaaaacaccccaattggtgatggtccagttctgctgccagataaccactacctgtctacccaatctgcactgtctaaagatccaaacgaaaaacgtgatcacatggtgctgctggaatttcgtaccgcagcaggtattacccacggtatggatgaactgtacaaaTAAATGT +mTurquoise(Vn)_ctag,pOpen_v3,dna,ATCCatggtttctaaaggtgaagaactgtttaccggtgttgttccaattctggttgaactggatggtgatgttaacggtcacaaattttctgtttctggtgaaggcgaaggtgatgcaacctacggtaaactgaccctgaaatttatctgtaccactggtaaactgccagttccatggccaactctggttaccactctgtcttggggtgttcaatgttttgcacgttacccagatcacatgaaacaacacgattttttcaaaagcgcaatgccagaaggttacgttcaagaacgtaccatcttcttcaaagatgacggtaactacaaaacccgtgcggaagtgaaatttgaaggtgataccctggttaaccgtatcgaactgaaaggtatcgacttcaaagaggatggcaacattctgggtcacaaactggaatacaactactttagcgataacgtgtacatcaccgcagataaacaaaaaaacggtatcaaggcgaacttcaaaatccgtcacaacattgaagatggtggtgttcaactggcagatcactaccaacaaaacactccaattggtgatggtccagttctgctgccagataaccactacctgtctacccaatctaaactgtctaaagacccaaacgaaaaacgtgatcacatggttctgctggaatttgttaccgcagcaggtattaccttaggtatggatgaactgtacaaaTAAATGT +sfGFP(Vn)_ctag,pOpen_v3,dna,ATCCatgcgtaaaggtgaagaactgtttaccggtgttgttccaattctggttgaactggatggtgatgttaacggtcacaaattttctgttcgtggtgaaggcgaaggtgatgcaaccaacggtaaactgaccctgaaatttatctgtaccactggtaaactgccagttccatggccaactctggttaccactctgacctacggtgttcaatgttttgcacgttacccagatcacatgaaacaacacgattttttcaaaagcgcaatgccagaaggttacgttcaagaacgtaccatctcttttaaagatgacggcacctacaaaacccgtgcggaagttaaatttgaaggtgataccctggttaaccgcattgaactgaaaggcatcgattttaaagaagatggtaacatcctgggccacaaactggaatacaactttaactctcacaacgtgtacatcaccgcagacaaacaaaaaaacggtatcaaagcgaacttcaagatccgtcacaacgttgaagatggttctgttcaactggcagatcactaccaacaaaacaccccaattggtgatggtccagttctgctgccagataaccactacctgtctacccaaagcgttctgtctaaagatccaaacgaaaaacgtgatcacatggtgctgctggaatttgttaccgcagcaggtattacccacggtatggatgaactgtacaaagcaTAAATGT +mVenus(Vn)_ctag,pOpen_v3,dna,ATCCatggtttctaaaggtgaagaactgtttaccggtgttgttccaattctggttgaactggatggtgatgttaacggtcacaaattttctgtttctggtgaaggcgaaggtgatgcaacctacggtaaactgaccctgaaactgatttgtaccactggtaaactgccagttccatggccaactctggttaccaccttaggttacggtctgcaatgttttgcacgttacccagatcacatgaaacaacacgattttttcaaaagcgcaatgccagaaggttacgttcaagaacgtaccatcttcttcaaagatgacggtaactacaaaacccgtgcggaagtgaaatttgaaggtgataccctggttaaccgtatcgaactgaaaggtatcgacttcaaagaggatggcaacattctgggtcacaaactggaatacaactacaactctcacaacgtttacatcaccgcagataaacaaaaaaacggcatcaaagcgaacttcaaaatccgtcacaacattgaagatggtggtgttcaactggcagatcactaccaacaaaacaccccaattggtgatggtccagttctgctgccagataaccactacctgtcttaccaatctaaactgagcaaagacccaaacgaaaaacgtgatcacatggttctgctggaatttgttaccgcagcaggtattaccttaggtatggatgaactgtacaaaTAAATGT +mScarlet-I(Vn)_ctag,pOpen_v3,dna,ATCCatggtttctaaaggtgaagcagtgatcaaagaatttatgcgcttcaaagttcacatggaaggttctatgaacggccacgaatttgaaattgaaggtgaaggcgaaggtcgtccatacgaaggtactcaaaccgcaaaactgaaagttaccaaaggtggtccactgccattttcttgggatattctgtctccacaatttatgtacggttctcgtgcatttatcaaacacccagcagatattccagactactacaaacaatcttttccggaaggtttcaaatgggaacgtgttatgaattttgaagatggtggtgcagttacggttacccaagatacctctctggaagatggtactctgatctacaaagttaaactgcgtggtactaactttccaccagatggtccagttatgcagaaaaaaaccatgggttgggaagcatctaccgaacgtctgtacccagaagatggcgttctgaaaggtgatatcaaaatggcactgcgtctgaaagatggcggtcgttacctggcagatttcaaaaccacctacaaagcgaaaaaaccagttcaaatgccaggtgcatacaacgttgatcgtaaactggatattaccagccacaacgaagattacaccgttgttgaacaatacgaacgttctgaaggccgtcactctaccggtggtatggatgaactgtacaaaTAAATGT +mCherry_ctag,pOpen_v3,dna,ATCCatggtttctaaaggtgaagaggataacatggcgatcatcaaagaatttatgcgcttcaaagttcacatggaaggttctgttaacggccacgaatttgaaattgaaggtgaaggcgaaggtcgtccatacgaaggtactcaaaccgcaaaactgaaagttaccaaaggtggtccactgccatttgcatgggatattctgtctccacagtttatgtacggtagcaaagcatacgttaaacacccagcagatattccagattacctgaaactgtcttttccggaaggtttcaaatgggaacgtgttatgaattttgaagatggtggtgttgttacggttacccaagattcttctctgcaagatggtgagtttatctacaaagttaaactgcgtggcaccaactttccatctgatggtccagttatgcagaaaaaaaccatgggttgggaagcatcttctgaacgtatgtacccagaagatggcgcactgaaaggtgaaattaaacaacgtctgaaacttaaagatggcggtcactacgatgcagaagttaaaaccacctacaaagcgaaaaaaccagttcaactgccaggtgcatacaacgttaacattaaactggatatcaccagccacaacgaagattacaccattgttgaacaatacgaacgtgcagaaggccgtcactctaccggtggtatggatgaactgtacaaaTAAATGT +mKate-2_ctag,pOpen_v3,dna,ATCCatggtttctgaactgattaaagaaaacatgcacatgaaactgtacatggaaggtactgttaacaaccaccacttcaaatgtacctctgaaggtgaaggtaaaccatacgaaggtactcaaaccatgcgtattaaagcagttgaaggtggtccactgccatttgcatttgatattctggcaacctcttttatgtacggcagcaaaacctttatcaaccacactcaaggtatcccggattttttcaaacaaagctttccagaaggtttcacctgggaacgtgttaccacctacgaagatggtggtgttctgaccgcaactcaagatacctctctgcaagatggttgtctgatctacaacgttaaaatccgtggtgttaactttccatctaacggtccagttatgcagaaaaaaaccttaggttgggaagcatctaccgaaactctgtacccagcggatggtggtctggaaggtcgtgcagatatggcactgaaactggttggtggtggtcacctgatttgtaacctgaaaaccacctaccgttctaaaaaaccagcgaaaaatctgaaaatgccaggtgtttactacgttgatcgtcgtctggaacgtatcaaagaagcagataaagaaacctacgtggaacaacacgaagttgcagttgcacgttactgtgatctgccatctaaactgggtcaccgtTAAATGT +I11012_ctag,pOpen_v3,dna,ATCCgcagcaaacgacgaaaactacgctgctgctgttTAAATGT +M0050_ctag,pOpen_v3,dna,ATCCgctgctaacgacgaaaactacgctctggctgctTAAATGT +M0051_ctag,pOpen_v3,dna,ATCCgctgctaacgacgaaaactacaactacgctgacgcttctTAAATGT +M0052_ctag,pOpen_v3,dna,ATCCgctgctaacgacgaaaactacgctgacgcttctTAAATGT +PARS,pOpen_v3,dna,GTCTcgataagctgggggaacattcgcgaaaatgaaacaagtcggctgttatagtatatttattataatattgaaagatctcaaaagactacttatttttgaatgaaccaagtatgaaatcaacctatttggggttgaccaaaataagtaaatattaattggatccgcacaaacgaaggAAGC +Pichia_Bxb1,pOpen_v3,dna,GTCTtggccgtggccgtgctcgtcctcgtcggccggcttgtcgacgacggcggtcaccgtcgtcaggatcatccgggccacaagcttgctgacagaagcctcaagaaaaaaaaaattcttcttcgactatgctggaggcagagatgatcgagccggtagttaactatatatagctaaattggttccatcacAAGC +tAOX1_terminator,pOpen_v3,dna,ATCCTAAtcaagaggatgtcagaatgccatttgcctgagagatgcaggcttcatttttgatacttttttatttgtaacctatatagtataggattttttttgtcattttgtttcttctcgtacgagcttgctcctgatcagcctatctcgcagctgatgaatatcttgtggtaggggtttgggaaaatcattcgagtttgatgtttttcttggtatttcccactcctcttcagagtacagaagattaagtgagaCGCT +tAOX1_terminator_ctag,pOpen_v3,dna,ATGTtcaagaggatgtcagaatgccatttgcctgagagatgcaggcttcatttttgatacttttttatttgtaacctatatagtataggattttttttgtcattttgtttcttctcgtacgagcttgctcctgatcagcctatctcgcagctgatgaatatcttgtggtaggggtttgggaaaatcattcgagtttgatgtttttcttggtatttcccactcctcttcagagtacagaagattaagtgagaCGCT +RFP(Ppastoris),pOpen_v3,dna,AatggcaacttccggtatggtgtcaaagggagaggaaaataatatggctattattaaggagtttatgcgttttaaggtacatatggaaggttctgtcaacggtcacgaattcgaaattgaaggtgagggggaggggaggccatacgagggaactcagactgctaagttaaaggtcactaaaggtggtcctttacctttcgcctgggatatcctgtctccacagtttatgtacggttcaaaggcttatgtgaaacatcctgccgatatcccagattatcttaaactttctttccctgagggttttaagtgggagagggtaatgaactttgaagacggtggtgtggtcactgttactcaggactcaagtctgcaggacggtgagttcatctacaaggtgaagctgagaggtaccaattttccatcagatggtcccgtgatgcaaaaaaagacaatgggttgggaagcttctagtgaacgtatgtatcccgaagatggagctttgaaaggtgaaattaagcaaagactaaaacttaaggatggtggacattacgatgctgaagttaagacgacctacaaggccaaaaagccagtccagttgcctggagcatacaatgttaacatcaaattggatataacttcccataatgaagactataccatcgtcgagcaatacgaacgagccgaagggagacacagtactggtggtatggatgaactttataaaggatccggaaccgcaGGATCC +EGFP(Ppastoris),pOpen_v3,dna,AatggtgagcaagggcgaggagctgttcaccggggtggtgcccatcctggtcgagctggacggcgacgtaaacggccacaagttcagcgtgtccggcgagggcgagggcgatgccacctacggcaagctgaccctgaagttcatctgcaccaccggcaagctgcccgtgccctggcccaccctcgtgaccaccctgacctacggcgtgcagtgcttcagccgctaccccgaccacatgaagcagcacgacttcttcaagtccgccatgcccgaaggctacgtccaggagcgcaccatcttcttcaaggacgacggcaactacaagacccgcgccgaggtgaagttcgagggcgacaccctggtgaaccgcatcgagctgaagggcatcgacttcaaggaggacggcaacatcctggggcacaagctggagtacaactacaacagccacaacgtctatatcatggccgacaagcagaagaacggcatcaaggtgaacttcaagatccgccacaacatcgaggacggcagcgtgcagctcgccgaccactaccagcagaacacccccatcggcgacggccccgtgctgctgcccgacaaccactacctgagcacccagagcgccctgagcaaagaccccaacgagaagcgcgatcacatggtcctgctggagttcgtgaccgccgccgggatcactctcggcatggacgagctgtacaagGGATCC +ntag_pichia_alphaAmylase-alphaMFDelta,pOpen_v3,dna,AACTatggtggcatggtggtccttattcttatatggtcttcaagttgctgctcctgcccttgctatgagatttcctagtattttcactgctgtgctatttgccgctagttccgctctagctgctccagttaatactactactgaagatgaattggagggtgacttcgatgttgctgttctgcctttttccgcttctatcgcagccaaggaagaaggtgtatctctagagaagcgtggAATG +ntag_pichia_killer-alphaMFDelta,pOpen_v3,dna,AACTatgaccaaaccaacgcaagtcttagttcgttcagtctctattttattcttcatcacactgttgcacttggttgttgcaatgagatttcctagtattttcactgctgtgctatttgccgctagttccgctctagctgctccagttaatactactactgaagatgaattggagggtgacttcgatgttgctgttctgcctttttccgcttctatcgcagccaaggaagaaggtgtatctctagagaagcgtggAATG +ntag_pichia_invertase-alphaMFDelta,pOpen_v3,dna,AACTatgttattgcaagcttttttatttctgctggcaggttttgcagcaaagatttctgccatgagatttcctagtattttcactgctgtgctatttgccgctagttccgctctagctgctccagttaatactactactgaagatgaattggagggtgacttcgatgttgctgttctgcctttttccgcttctatcgcagccaaggaagaaggtgtatctctagagaagcgtggAATG +ntag_pichia_inulinase-alphaMFDelta,pOpen_v3,dna,AACTatgaaactggcttactccctgttgctacctctggctggagtttccgctatgagatttcctagtattttcactgctgtgctatttgccgctagttccgctctagctgctccagttaatactactactgaagatgaattggagggtgacttcgatgttgctgttctgcctttttccgcttctatcgcagccaaggaagaaggtgtatctctagagaagcgtggAATG +ntag_pichia_serumalbumin-alphaMFDelta,pOpen_v3,dna,AACTatgaagtgggtaactttcatctcattgttattcttgttctcctctgcttactctatgagatttcctagtattttcactgctgtgctatttgcctctagttccgctctagctgctccagttaatactactactgaagatgaattggagggtgacttcgatgttgctgttctgcctttttccgcttctatcgcagccaaggaagaaggtgtatctctagagaagcgtggAATG +ntag_pichia_glucoamylase-alphaMFDelta,pOpen_v3,dna,AACTatgtctttcagatccctattggcattgtcagggttggtctgttctggattggctatgagatttcctagtattttcactgctgtgctatttgccgctagttccgctctagctgctccagttaatactactactgaagatgaattggagggtgacttcgatgttgctgttctgcctttttccgcttctatcgcagccaaggaagaaggtgtatctctagagaagcgtggAATG +ntag_pichia_alphaMFDelta-noKex,pOpen_v3,dna,AACTatgagattcccatcaatttttactgctgttctgttcgccgcttctagtgcacttgccatgagatttcctagtattttcactgctgtgctatttgccgctagttccgctctagctgctccagttaatactactactgaagatgaattggagggtgacttcgatgttgctgttctgcctttttccgcttctatcacagccaaggaagaaggtgtatctctagagaagcgtggAATG +ntag_pichia_alphaMFDelta,pOpen_v3,dna,AACTatgagatttcctagtattttcactgctgtgctatttgccgctagttccgctctagctgctccagttaatactactactgaagatgaattggagggtgacttcgatgttgctgttctgcctttttccgcttctatcgcagccaaggaagaaggtgtatctctagagaagcgtggAATG +ntag_pichia_alphaMF-noEAEA,pOpen_v3,dna,AACTatgagatttccttcaatttttactgctgttttattcgcagcatcctccgcattagctgctccagtcaacactacaacagaagatgaaacggcacaaattccggctgaagctgtcatcggttactcagatttagaaggggatttcgatgttgctgttttgccattttccaacagcacaaataacgggttattgtttataaatactactattgccagcattgctgctaaagaagaaggggtatctctcgagaaaagaggAATG +ntag_pichia_alphaMF,pOpen_v3,dna,AACTatgagatttccttcaatttttactgctgttttattcgcagcatcctccgcattagctgctccagtcaacactacaacagaagatgaaacggcacaaattccggctgaagctgtcatcggttactcagatttagaaggggatttcgatgttgctgttttgccattttccaacagcacaaataacgggttattgtttataaatactactattgccagcattgctgctaaagaagaaggggtatctctcgagaaaagagaggctgaagctggAATG +pTPI1_promoter,pOpen_v3,dna,TACAgtgtttaaagattacggatatttaacttacttagaataatgccatttttttgagttataataatcctacgttagtgtgagcgggatttaaactgtgaggaccttaatacattcagacacttctgcggtatcaccctacttattcccttcgagattatatctaggaacccatcaggttggtggaagattacccgttctaagacttttcagcttcctctattgatgttacacctggacaccccttttctggcatccagtttttaatcttcagtggcatgtgagattctccgaaattaactaaagcaatcacacaattctctcggataccacctcggttgaaactgacaggtggtttgttacgcatgctaatgcaaaggagcctatatacctttggctcggctgctgtaacagggaatataaagggcagcataatttaggagtttagtgaacttgcaacatttactattttcccttcttacgtaaatatttttctttttaattctaaatcaatctttttcaattttttgtttgtattcttttcttgcttaaatctataactacaaaaaacacatacataaactaaaaAACT +pENO1_promoter,pOpen_v3,dna,TACAagaaagcatactatactattcgacattcctttcaatcctggaattaacagtcacttttaaaaaagacatctaccgtgaaggtgccgtagagtatcgcgttaccatatcgccaaaaactgatatacgccgcggaaaccaggcaaacaattgaaaagaaaaattttgaggaactctctgcatcgaagccgtctagagttaccactagtcagatgccgcgggcacttgagcacctcatgcacagcaataacacaacacaatggttagtagcaacctgaattcggtcattgatgcatgcatgtgccgtgaagcgggacaaccagaaaagtcgtctataaatgccggcacgtgcgatcatcgtggcggggttttaagagtgcatatcacaaattgtcgcattaccgcggaaccgccagatattcattacttgacgcaaaagcgtttgaaataatgacgaaaaagaaggaagaaaaaaaaagaaaaataccgcttctaggcgggttatctactgatccgagcttccactaggatagcacccaaacacctgcatatttggacgacctttacttacaccaccaaaaaccactttcgcctctcccgcccctgataacgtccactaattgagcgattacctgagcggtcctcttttgtttgcagcatgagacttgcatactgcaaatcgtaagtagcaacctctcaaggtcaaaactgtatggaaaccttgtcacctcacttaattctagctagcctaccctgcaagtcaagagctctccgtgattcctagccacctcaaggtatgcctctccccggaaactgtggccttttctggcacacatgatctccacgatttcaacatataaatagcttttgataatggcaatattaatcaaatttattttacttctttcttgtaacatctctcttgtaatcccttattccttctagctatttttcataaaaaaccaagcaactgcttatcaacacacaaacactaaatcaaaAACT +pAOX1_promoter,pOpen_v3,dna,TACAgatctaacatccaaagacgaaaggttgaatgaaacctttttgccatccgacatccacaggtccattctcacacataagtgccaaacgcaacaggaggggatacactagcagcagaccgttgcaaacgcaggacctccactcctcttctcctcaacacccacttttgccatcgaaaaaccagcccagttattgggcttgattggagctcgctcattccaattccttctattaggctactaacaccatgactttattagcctgtctatcctggcccccctggcgaggttcatgtttgtttatttccgaatgcaacaagctccgcattacacccgaacatcactccagatgagggctttctgagtgtggggtcaaatagtttcatgttccccaaatggcccaaaactgacagtttaaacgctgtcttggaacctaatatgacaaaagcgtgatctcatccaagatgaactaagtttggttcgttgaaatgctaacggccagttggtcaaaaagaaacttccaaaagtcggcataccgtttgtcttgtttggtattgattgacgaatgctcaaaaataatctcattaatgcttagcgcagtctctctatcgcttctgaaccccggtgcacctgtgccgaaacgcaaatggggaaacacccgctttttggatgattatgcattgtctccacattgtatgcttccaagattctggtgggaatactgctgatagcctaacgttcatgatcaaaatttaactgttctaacccctacttgacagcaatatataaacagaaggaagctgccctgtcttaaacctttttttttatcatcattattagcttactttcataattgcgactggttccaattgacaagcttttgattttaacgacttttaacgacaacttgagaagatcaaaaaacaactaattattcgaaacgAACT +PxylA,pOpen_v3,dna,TACAaaggccaaaaaactgctgccttcggatcagcgatatccacttcatccactccatttgtttaatctttaaattaagtatcaacatagtacatagcgaatcttccctttattatatctaatgtgttcataaaaaactaaaaaaaatattgaaaatactgacgaggttatataagatgaaaataagttagtttgtttaaacaacaaactaataggtgatgtacttactatatgaaataaaatgcatctgggatcccaagcttatcgaaAACT +PliaI,pOpen_v3,dna,TACAattggccaaagcagaaaggtccgacctaattaaagaaagggaagcaagtgttcatctgtaaagggttttaaaacgccatgcctcgtgcatggcgtttttttgtgccaatgggtccggtgcgagatacgactccggtcttatataaaaatcaatctctgattcgttttgcatatcttccaacttgtataagatgaagacaaggaaaacgaAACT +P_J23101,pOpen_v3,dna,TACAtttacagctagctcagtcctaggtattatgctagcAACT +PlepA,pOpen_v3,dna,TACAagtcaatgtatgaatggatacgggatatgaatcaataagtacgtgaaagagaaaagcaacccagatatgatagggaacttttctctttcttgttttacattgaatctttacaatcctattgatataatctaagctagtgtattttgcgtttaatagtAACT +PliaG,pOpen_v3,dna,TACAcaaaaatcagaccagacaaaagcggcaaatgaataagcggaacggggaaggatttgcggtcaagtccttcccttccgcacgtatcaattcgcaagcttttcctttataatagaatgaatgaAACT +Pveg,pOpen_v3,dna,TACAggagttctgagaattggtatgccttataagtccaattaacagttgaaaacctgcataggagagctatgcgggttttttattttacataatgatacataatttaccgaaacttgcggaacataattgaggaatcatagaattttgtcaaaataattttattgacaacgtcttattaacgttgatataatttaaattttatttgacaaaaatgggctcgtgttgtacaataaatgtagtAACT +US_amyE,pOpen_v3,dna,ATTAttaacaaaattctccagtcttcacatcggtttgaaaggaggaagcggaagaatgaagtaagagggatttttgactccgaagtaagtcttcaaaaaatcaaataaggagtgtcaagaatgtttgcaaaacgattcaaaacctctttactgccgttattcgctggatttttattgctgtttcatttggttctggcaggaccggcggctgcgagtgctgaaacggcgaacaaatcgaatgagcttacagcaccgtcgatcaaaagcggaaccattcttcatgcatggaattggtcgttcaatacgttaaaacacaatatgaaggatattcatgatgcaggatatacagccattcagacatctccgattaaccaagtaaaggaagggaatcaaggagataaaagcatgtcgaactggtactggctgtatcagccgacatcgtatcaaattggcaaccgttacttaggtactgaacaagaatttaaagaaatgtgtgcagccgctgaagaatatggcataaaggtcattgttgacgcggtcatcaatcataccaccagtgattatgccgcgatttccaatgaggttaagagtattccaaactggacacatggaaacacacaaattaaaaactggtctgatcCGAG +DS_amyE,pOpen_v3,dna,GTCTagagtcgacatggatgagcgatgatgatatccgtttaggctgggcggtgatagcttctcgttcaggcagtacgcctcttttcttttccagacctgagggaggcggaaatggtgtgaggttcccggggaaaagccaaataggcgatcgcgggagtgctttatttgaagatcaggctatcactgcggtcaatagatttcacaatgtgatggctggacagcctgaggaactctcgaacccgaatggaaacaaccagatatttatgaatcagcgcggctcacatggcgttgtgctggcaaatgcaggttcatcctctgtctctatcaatacggcaacaaaattgcctgatggcaggtatgacaataaagctggagcgggttcatttcaagtgaacgatggtaaactgacaggcacgatcaatgccaggtctgtagctgtgctttatcctgatgatattgcaaaagcgcctcatgttttccttgagaattacaaaacaggtgtaacacattctttcaatgatcaactgacgattaccttgcgtgcagatgcgaatacaacaaaagccgtttatcaaatcaataatggaccagagacggcgtttaaggatggagatcaattcacaatcggaaaaggagatccatttggcaaaacatacaccatcatgttaaaaggaacgaacagtgatggtgtaacgaggaccgagaaatacagttttgttaaaagagatccagcgtcggccaaaaccatcggctatcaaaatccgaatcattggagccaggtaaatgcttatatctataaacatgatgggagccgagtaattgaattgaccggatcttggcctggaaaaccaatgactaaaaatgcagacggaatttacacgctgacgctgcctgcggacacggatacaaccaacgcaaaagtgatttttaataatggcagcgcccaagtgcccggtcagaatcagcctggctttgattacgtgctaaatggtttatataatgactcgggcttaagcggttctcttccccattgagggcaaggctagacgggacttaccgaaagaaaccatcaatgatggtttcttttttgttcataaatcagacaaaacttttctcttgcaaaagtttgtgaagtgttgcacaatataaatgtgaaatacttcacaaacaaaaagacatcaaagagaaacataccctgAAGC +camR(Bsub),pOpen_v3,dna,AAGCtccggcaaaaaagggcaaggtgtcaattctcatgtttgacagcttatcatcggcaatagttacccttattatcaagataagaaagaaaaggatttttcgctacgctcaaatcctttaaaaaaacacaaaagaccacattttttaatgtggtctttattcttcaactaaagcacccattagttcaacaaacgaaaattggataaagtgggatatttttaaaatatatatttatgttacagtaatattgacttttaaaaaaggattgattctaatgaagaaagcagacaagtaagcctcctaaattcactttagataaaaatttaggaggcatatcaaatgaactttaataaaattgatttagacaattggaagagaaaagagatatttaatcattatttgaaccaacaaacgacttttagtataaccacagaaattgatattagtgttttataccgaaacataaaacaagaaggatataaattttaccctgcatttattttcttagtgacaagggtgataaactcaaatacagcttttagaactggttacaatagcgacggagagttaggttattgggataagttagagccactttatacaatttttgatggtgtatctaaaacattctctggtatttggactcctgtaaagaatgacttcaaagagttttatgatttatacctttctgatgtagagaaatataatggttcggggaaattgtttcccaaaacacctatacctgaaaatgctttttctctttctattattccatggacttcatttactgggtttaacttaaatatcaataataatagtaattaccttctacccattattacagcaggaaaattcattaataaaggtaattcaatatatttaccgctatctttacaggtacatcattctgtttgtgatggttatcatgcaggattgtttatgaactctattcaggaattgtcagataggcctaatgactggcttttataatatgagataatgccgactgtactttttacagtcggttttctaatgtcactaacctgccccgttagttgaagaaggtttttatattacagctccagatcctctacgccggacgcatcgtggccggcatcaccggcgccacaggtgcggttgctggcgcctatatcgccgacatcaccgatggggaagatcgggctcgccacttcgggctcatgagcgcttgtttcggcgtgggtatggtggcaggccccgtggccgggggactgttgggcgccatctccttgcatgcaccattccttgcggcggcggtgctcaacggcctcaacctactactgggctgcttcctaatgcaggagtcgcataagggagATAG +US_lacA,pOpen_v3,dna,ATTAgtgatgtcaaagcttgaaaaaacgcacgtaacaaaagcaaaatttatgctccatgggggagactacaaccccgatcagtggctggatcggcccgatattttagctgacgatatcaaactgatgaagctttctcatacgaatacgttttctgtcggcatttttgcatggagcgcacttgagccggaggagggcgtatatcaatttgaatggctggatgatatttttgagcggattcacagtataggcggccgggtcatattagcaacgccgagcggagcccgtccggcctggctgtcgcaaacctatccggaagttttgcgcgtcaatgcctcccgcgtcaaacagctgcacggcggaaggcacaaccactgcctcacatctaaagtctaccgagaaaaaacacggcacatcaaccgcttattagcagaacgatacggacatcacccggcgctgttaatgtggcacatttcaaacgaatacgggggagattgccactgtgaCGAG +DS_lacA,pOpen_v3,dna,GTCTttcaagctatatttggagttgagcctcttgaaacggacaccctgtatccgaaggatcgaaacgctgtcagctaccgcagccaaatatatgaaatgaaggattatgcaaccgtgattgatgtaaagacagcttcagtggaagcggtgtatcaagaagatttttatgcgcgcacgccagcggtcacaagccatgagtatcagcagggcaaggcgtattttatcggcgcgcgtttggaggatcaatttcagcgtgatttctatgagggtctgatcacagacctgtctctctctccagtttttccggttcggcacggaaaaggcgtctccgtacaagcgaggcaggatcaggacaatgattatatttttgtcatgaatttcacggaagaaaaacagctggtcacgtttgatcagagtgtgaaggacataatgacaggagacatattgtcaggcgacctgacgatggaaaagtatgaagtgagaattgtcgtaaacacacattagAAGC +eryR(Bsub),pOpen_v3,dna,AAGCgagttaacaagagtttgtagaaacgcaaaaaggccatccgtcaggatggccttctgcttagctagagcggcggatttgtcctactcaggagagcgttcaccgacaaacaacagataaaacgaaaggcccagtctttcgactgagcctttcgttttatttgatgcctcaagctagagagtcctagagtctagggacctctttagctccttggaagctgtcagtagtatacctaataatttatctacattccctttagtaacgtgtaactttccaaatttacaaaagcgactcatagaattatttcctcccgttaaataatagataactattaaaaatagacaatacttgctcataagtaacggtacttaaattgtttactttggcgtgtttcattgcttgatgaaactgatttttagtaaacagttgacgatattctcgattgacccattttgaaacaaagtacgtatatagcttccaatatttatctggaacatctgtggtatggcgggtaagttttattaagacactgtttacttttggtttaggatgaaagcattccgctggcagcttaagcaattgctgaatcgagacttgagtgtgcaagagcaaccctagtgttcggtgaatatccaaggtacgcttgtagaatccttcttcaacaatcagatagatgtcagacgcatggctttcaaaaaccacttttttaataatttgtgtgcttaaatggtaaggaatactcccaacaattttatacctctgtttgttagggaattgaaactgtagaatatcttggtgaattaaagtgacacgagtattcagttttaatttttctgacgataagttgaatagatgactgtctaattcaatagacgttacctgtttacttattttagccagtttcgtcgttaaatgccctttacctgttccaatttcgtaaacggtatcggtttcttttaaattcaattgttttattatttggttgagtactttttcactcgttaaaaagttttgagaatattttatatttttgttcatgtaatcactccttcttaattacaaatttttagcatctaatttaacttcaattcctattatacaaaattttaagatactgcactatcaacacactcttaagtttgcttctaagtcttatttccataacttcttttacgtttccgccattctttgctgtttcgatttttatgatatggtgcaagtcagcacgaacacgaaccgtcttatctcccattatatctttttttgcactgattggtgtatcatttcgtttttcttttgtgctagaggatcaattcttgaagacgaaagggcctcgtgatacgcctatttttataggttaatgtcatgataataatggtttcttagacgtcaggtggcacttttcggggaaatgtgATAG +US_thrC,pOpen_v3,dna,ATTAaattcatgtaaaagatgaggttggttcattctcgaaaattacatctgtgttctcagagcggggcgtgagctttgaaaaaatccttcagctgccaattaaaggccatgatgagttagctgaaatcgtaattgtcacacatcatacatcagaagctgatttcagtgatatcctgcaaaacctaaatgatttggaagtcgttcaagaagtcaaaagcacatatcgtgtagaagggaacggttggagctaatgtggaaaggacttatccatcaatataaagaatttttacctgtaacagatcaaacaccggcgctaactttacatgaaggaaacacacctcttattcacctgccgaagctgtctgagcagctcggaattgagcttcatgtcaaaacggaaggcgtcaatcctacgggatcatttaaagatcgcggaatggttatggctgtggcaaaggcaaaagaagaaggcaatgacacgattatgtgcgcgtcaacaggtaacacttccgctgctgcggcagcatatgcagcccgtgctaacatgaaatgcattgtcatcatcccgaacggaaaaattgcatttggaaaactcgctcaagctgtcatgtacggagccgagattatcgcaattgacggaaactttgacgatgcgcttaaaattgtccgttccatctgtgagaaatcaccgattgcccttgtcaactcagtcaacccttaccgcattgaaggccaaaaaactgctgccttcgCGAG +DS_thrC,pOpen_v3,dna,GTCTaggccttccgaaaatgcgcggctttgaagctgagggtgcggcggcaatcgtgcgcaatgaagtgattgaaaatccggaaacaatagcgacagccattcgtatcggaaacccggcaagctgggacaaagctgtaaaggcagccgaggaatccaatgggaaaattgacgaagtcactgatgatgaaatccttcacgcatatcagctgatcgcccgtgtagaaggcgtgtttgcagaaccaggttcttgcgcgtctatcgcaggagtgctgaaacaggtgaaatccggagaaattccgaaaggcagcaaggtcgtagctgtgttaacaggaaacggactgaaagatccgaacacagcggtcgacatttcagaaatcaagcctgtcacattgccgactgatgaagacagcatccttgaatatgtaaaaggagcggcccgtgtatgaacgaagccgacatgctgttctctgtcactgttcccggaagcacagctaacctaggccccggctttgattcagtcggaatggcgctcagcagatatttgaagctgaccgtctttgaaagcgacaaatggtcttttgaggctgaaacagaaacagtcgccggaattAAGC +Pars,pOpen_v3,dna,TACAttaatcaaaataaattgatttatttgcttgcattaatttaaaaatcatgagtataataaatacatcaaaaAACT +P43,pOpen_v3,dna,TACAtattttacatttttagaaatgggcgtgaaaaaaagcgcgcgattatgtaaaatataaAACT +PxylA_xylR,pOpen_v3,dna,TACAagcggataacaatttcacacaggaggccgatcgcgttatgcagcaatggcaagaacgtcccggggagctcctaacttataggggtaacacttaaaaaagaatcaataacgatagaaaccgctcctaaagcaggtgcattttttcctaacgaagaaggcaatagttcacatttattgtctaaatgagaatggactctagaagaaacttcgtttttaatcgtatttaaaacaatgggatgagattcaattatatgatttctcaagataacagcttctatatcaaatgtattaaggatattggttaatccaattccgatataaaagccaaagttttgaagtgcatttaacatttctacatcatttttatttgcgcgttccacaatctcttttcgagaaatattcttttcttctttagagagcgaagccagtaacgctttttcagaagcatataattcccaacagcctcgatttccacagctgcatttgggtccattaaaatctatcgtcatatgacccatttccccagaaaaaccctgaacacctttatacaattcgttgttaataacaagtccagttccaattccgatattaatactgatgtaaacgatgttttcatagttttttgtcataccaaatactttttcaccgtatgctcctgcattagcttcattttcaacaaaaaccggaacattaaactcactctcaattaaaaactgcaaatctttgatattccaatttaagttaggcatgaaaataatttgctgatgacgatctacaaggcctggaacacaaattcctattccgactagaccataaggggactcaggcatatgggttacaaaaccatgaataagtgcaaataaaatctcttttacttcactagcggaagaactagacaagtcagaagtcttctcgagaataatatttccttctaagtcggttagaattccgttaagatagtcgactcctatatcaataccaatcgagtagcctgcattcttattaaaaacaagcattacaggtcttctgccgcctctagattgccctgccccaatttcaaaaataaaatctttttcaagcagtgtatttacttgagaggagacagtagacttgtttaatcctgtaatctcagagagagttgccctggagacaggggagttcttcaaaatttcatctaatattaatttttgattcattttttttactaaagcttgatctgcaatttgaataataaccactcctttgtttatccaccgaactaagttggtgttttttgaagcttgaattagatatttaaaagtatcatatctaatattataactaaattttctaaaaaaaacattgaaataaacatttattttgtatatgatgagataaagttagtttattggataaacaaactaactcaattaagatagttgatggataaacttgttcacttaaatcaaaggctgagAACT +US_glmS,pOpen_v3,dna,ATTAcaacatgccgcttctgtctaagaaaccgctcttcattttcctttctcaaagcggagaaacagcagacagccgcgcggtactcgttcaagtcaaagcgctcggacacaaagccctgacaatcacaaacgtacctggatcaacgctttctcgtgaagctgactatacattgctgcttcatgcaggccctgagatcgctgttgcgtcaacgaaagcatacactgcacaaatcgcagttctggcggttcttgcttctgtggctgctgacaaaaatggcatcaatatcggatttgacctcgtcaaagaactcggtatcgctgcaaacgcaatggaagctctatgcgaccagaaagacgaaatggaaatgatcgctcgtgaatacctgactgtatccagaaatgctttcttcatcggacgcggccttgactacttcgtatgtgtcgaaggcgcactgaagctgaaagagatttcttacatccaggcagaaggttttgccggcggtgagctaaagcacggaacgattgccttgatcgaacaaggaacaccagtattcgcactggcaactcaagagcatgtaaacctaagcatccgcggaaacgtcaaagaagttgctgctcgcggagcaaacacatgcatcatctcactgaaaggcctagacgatgcggatgacagattcgtattgccggaagtaaacccagcgcttgctccgttggtatctgttgttccattgcagctgatcgcttactatgctgcactgcatcgcggctgtgatgtggataaacctcgtaaccttgcgaagagtgttactgtggagtaataaatgtttaacccctttggataagattatctaaaggggtgtCGAG +DS_glmS,pOpen_v3,dna,GTCTttatgtccaaaagcgaaagaacgtctacagtggaaaaatggattaaagaagttgatggtacaggacccgactatcgatcgaccatggctatagatttaagatgtttcctcattagatcggtcaatcaggttaaaaggcattaaaacttggagagaaactgcttttgcctaactgagttttctaatgttaatacagatataagagagcagttcctcattaagatacgcttttgatagcagttggaatttagcatgctagaaatccgaaaaccggagagcctattggaatgataacgtgtttttttattgacaattaataaagggcaaagaattctagaagtcgcacttacaatgtacttgtgggattaggttttaaagttcattctttttttattgagtacattccggttaaagttgaaatgattcagtaaaggacacgccataaataaaaggtagcgtatgattgccattctttccatggaaccgatatttctaagatttcatctttcgtaggctttcggttcatattacgtaatatttttatcgaatgaataagacccacatcatcgattggaaaagctgtcgggaacctaaggcagcgcattagaacataattggctgtccaaggaccgattcctcgtattttaattaagtttttttcagcatctttaaagttcattttcattaatttttccctcgataattctccacttgccattaatctggctattccaatgatatattcactttttttcaccgtcattttaatatctgccaggtcggtaggggttaaccgtgcaattcgctcgtacggtgggaacacccaatactttttaccattccattcaatagaatcgccaaatgcttctacaaattgcttctttaaggagtacgcgaaggctaaAAGC +Pmtl-short,pOpen_v3,dna,TACAtcctgaaaccaggagcctttttatttttaaaaaattgtcacagtcatgtgccaaagtcctctttactttcaattgtatagggactgtaagcgttttaacatagagtcaaagggaagcatcattctctgaacatatttAACT +Ptet_tetR,pOpen_v3,dna,TACAttaagacccactttcacatttaagttgtttttctaatccgcatatgatcaattcaaggccgaataagaaggctggctctgcaccttggtgatcaaataattcgatagcttgtcgtaataatggcggcatactatcagtagtaggtgtttccctttcttctttagcgacttgatgctcttgatcttccaatacgcaacctaaagtaaaatgccccacagcgctgagtgcatataatgcattctctagtgaaaaaccttgttggcataaaaaggctaattgattttcgagagtttcatactgtttttctgtaggccgtgtacctaaatgtacttttgctccatcgcgatgacttagtaaagcacatctaaaacttttagcgttattacgtaaaaaatcttgccagctttccccttctaaagggcaaaagtgagtatggtgcctatctaacatctcaatggctaaggcgtcgagcaaagcccgcttattttttacatgccaatacaatgtaggctgctctacacctagcttctgggcgagtttacgggttgttaaaccttcgattccgacctcattaagcagctctaatgcgctgttaatcactttacttttatctaatctagacatcattaattcctcctttttgttgacattatatcattgatagagttatttgtcaaactagatttttatttcgatgccctggacttcatgaaaaactaaaaaaaatattgacactctatcattgatagagtataattaaaataagcttgatcgtagcgttaacagatctgagctcctgcagtaagctgagAACT +Pman_manR,pOpen_v3,dna,TACAttaaagtgtgaataataagatcttgtctcaactgtataccgaaatcagctcattaaaatcgctttttttaccataggttccggtaataaaggcatttttccctataacaaaaaaagcaaggaataatccctgcttttaataatccaaatgagataaaaatgtcatgacattggtttctttcgtcagcttctggacgaaggccggctgctcgctaagatatgaaagctcgctaaacagctgctttgtcatagtttgatcctcgtgtttgacagccagcataaaaacgagcgaaactttttcatttccccattcaagcggctcctttaatgtggcaatcgcgattgctgattgtttaataaatttagcgtttgcatgaggaatggcaatgcctgacccaatatttgtagccgacattttttccctcatcaccgcatgaaccgcatagtccttatcaacatagcctttctcaaacaaagctgtcgccagctgttcgattaacttgtagcgatgctccgcctcctgctgcaaaaaaaccaaaaaaggagtcgtattgttcagcatctgaaacgttttttgtttttggcggtgtgattcgcccagctgacgtataaacgcacttaatttcttctcttcgccaggctctaaaagcggagacacaacaatatggggaaccgttatattctccagcgcaatggtggatatgacaagatcaatatcctcatgctttttgatataatcctttaaatcggcctttgcaatacaagccattacagcgatttggtgatatttccgttcaatttttgtccgcagcaattgtgacatcccaatccccatgtggcagacgataaccgcctttttatgtgtctcgctgctgccctgcatccgttcgatcgctgcctgaaaatgcaatgtcaaatatgctgcctcttcctcaggaatatgaagatcaaaagtttgattgatgtcttccaaaacatcaatgatgaggtggaacaaataagggtacatctttttgatgtcattgagcataggatttgctacagaaagatcgtagctcagccgttgtagcacagtattgagatgaatgttcaaaccattgatcaaatcctgatccttgtgaaaatccatcattttcagctctgaaacacgattgattaaatgccctacaaccttaggaagcacagcattttcaaggttctcttctgtctgcaatggataacgaaccttaccgcccaatatatgtaatgttaagtacacggcttcttcttcagggaagcgaattgcaaaaaccggttcaagccgctgtaaacaagcaaatgtccattgatattctttcttcttttttactgccgccatttcttttggggacaatgaaatcggctgcttcattttgatgcggcggaccattaacaatgtatgcagcaacaaactttcaaacgtttcgtctgtgaaatagagggaatgctttttttgtagtgatttgatttctttcgtcacaaagtctacttcataatgtaaaaacttacttttaataaattggctggtaaacgctgtattgtgaatcaaatcagatattcttgccaatgcctttcttttatttctttcgttcccttcaacctttaatccaagccgctgcctagaaaccagtgtgagatcaaaacgcttcagccactcttctactgcgtataaatcctttttaatggaagatcgattgacaaaatgccgagcggcaatgtcttttgcggaaacaggcttcggattcattaataagtcataagcaatgtgcagcattctttccttatcgctccgctgcctagaagaaaaatgttcagtatgtaattgttggcttagccaggtcctttcctgttcttctatatgtaaataaacgcccaaccctggttttcgaataagctgtgcatgagaatgctcattgagataatcctcgatcactttcaaatcattccggattgttttttcagaacactgtaccctatctgcgaaatcttgcacaactaaatagtcatcaggttcagataacaacagatacaaaatctctttttgtcttgtatttatgtattccatctgtttttctatccttcctgtcagtatctacttgagcaaattgatagtataagtatactatataagaaaccgtttacattataaaacaaaatcatagagacaaaagtttcgttttttaccgaagcttccggaaaaaagcgatttttatactctttgagtttgctatgAACT +P3P,pOpen_v3,dna,TACAtcttttggaagaaaatatagggaaaatggtatttgttaaaaattctgaatatttatacaatatcatatgtttcacattgaaatatgctgtccagactgtccgctgtgtaaaaaataggaataaaggggggttgttattattttactgatatgtaaaatataatttgtataagaaaattcgaaacgtaagatgaaaccttagataaaagtgctttttttgttgcaattgaagaattattaatgttaagcttaattaaagataatatctttgaattgtaacgcccctcaaaagtaagaactacaaaaaaagaatacgttatatagaaatatgtttgaaccttcttcagattacaaatatattcggacggactctacctcaaatgcttatctaactatagaatgacatacaagcacaaccttgaaaatttgaaaatataactaccaatgaacttgttcatgtgaattatcgctgtatttaattttctcaattcaatatataatatgccaatacattgttacaagtagaaattaagacacccttgatagccttactatacctaacatgatgtagtattaaatgaatatgtaaatatatttatgataagaagcgacttatttataatcattacatatttttctattggaatgattaagattccaatagaatagtgtataaattatttatcttgaaaggagggatgcctaaaaacgaagaacattaaaaacatatatttgcaccgtctaatggatttatgaaaaatcattttatcagtttgaaaattatgtattatAACT +US_sigF,pOpen_v3,dna,ATTAgctgaacttggaggacctttcctttatggacagctcggggcttggcgttattttaggaagatacaagcaaattaagcaaattggcggagaaatggttgtttgcgctatctctcctgcggtgaagcgattgtttgatatgtcgggtctgtttaaaattatccgatttgaacaatctgaacagcaggcactcctgacactgggggtggcatcatgaaaaatgaaatgcaccttgagttttctgccctcagtcagaatgaatcgttcgcccgtgtgacagttgcttcatttatagctcagctggacccgacaatggatgaactgactgaaatcaaaacagtcgtgtcagaggctgtcacgaatgcgattatccatggatatgaagagaactgtgaagggaaagtttacatttcagtgacgctggaagatcatgtcgtatatatgactattcgtgatgaaggcttaggcattacagatcttgaagaagcccgtcagcctctatttacgactaagcctgagcttgagcgctctggaatgggctttaccattatggaaaatttcatggatgatgtcagtatcgattcatcgcctgaaatgggaacaacgattcgcttaacaaagcacttatcaaaaagcaaagcgctttgtaattaaggagatttgttatggattaCGAG +DS_sigF,pOpen_v3,dna,GTCTtctgcagtgcaggctagcttttttgtgcaaaagcgtggtaatttatggtcttttcgagcggatgaatgagaacaaaatcgaaccacatactacatatataaccaccgaaagatggtgatcaatgatggaacgacgaatatttatccggcttcgccaccgagtgctggcacatccaggggatattattaccgttggagatgccgcgcaaatagaagggcagcttcagctgaaaaagaaactttcggctatgccgctttatcaggtgagcgaaaaagataaaaatatcgtaattctggatatcatacaagtcctcagagccattcatttacaagacccgacaattgatgttcaaaccgtaggcggagcagaaaccattgttgaaattcagtatcgaaagcgaaatttatcaacggttctatttatcggtgtctggctgcttctgtttattggatcgtgtcttgccatcatgaactttcatgaggatgtaagcatgagagatgttcatatcgcactatatgaaatcataaccggagagaggaatgactatccatatttgcttcaaatcccatacagcatcggtttgggactggggatgatcgtgttttttaaccacatatttaaaaagcgcctaaatgaagagcccagcccgctggaggttgagatgtttaactatcagcttgatctcgatcaatatgtggccatgcatgagaatcaagaaaccataaaggatctgcatgatcgttagtgtattgttcatcatttttgtcgggctcggcggaggcataacggtgggggcgggtttcgttgcttttttaaccgtaatgggaatcattccgcggctgatgcagctcaccaaaacaatgagatttgttcaggcttatgaagcgAAGC +kanR(Bsub),pOpen_v3,dna,AAGCgaggtcatcgttcaaaatggtatgcgttttgacacatccactatatatccgtgtcgttctgtccactcctgaatcccattccagaaattctctagcgattccagaagtttctcagagtcggaaagttgaccagacattacgaactggcacagatggtcataacctgaaggaagatctgattgcttaactgcttcagttaagaccgaagcgctcgtcgtataacagatgcgatgatgcagaccaatcaacatggcacctgccattgctacctgcacagtcaaggatggtagaaatgttgtcggtccttgcacacgaatattacgccatttgcctgcatattcaaacagctcttctacgataagggcacaaatcgcatcgtggaacgtttgggcttctaccgatttagcagtttgatacactttctctaagtatccacctgaatcataaatcggcaaaatagagaaaaattgaccatgtgtaagcggccaatctgattccacctgagatgcataatctagtagaatctcttcgctatcaaaattcacttccaccttccactcaccggttgtccattcatggctgaactctgcttcctctgttgacatgacacacatcatctcaatatccgaatagggcccatcagtctgacgaccaagagagccataaacaccaatagccttaacatcatccccatatttatccaatattcgttccttaatttcatgaacaatcttcattctttcttctctagtcattattattggtccattcactattctcattcccctttcagataattttagatttgcttttctaaataagaatatttggagagcaccgttcttattcagctattaaacccattatatcgggtttttgaggggatttcaactgcagATAG +US_yhgE,pOpen_v3,dna,ATTAgacgggtctaaacaggtccaaagcggatcatcaaaacttgcagacggatctgcatcacttgatacaggtttaggcaagctattggacggcacaggagagctgtctagtaagctgaaggatgcggctgaccaaacaggagatatcgatgctgatgatcaaacatacggcatgtttgcggatccggttaaaaccaaagatgatgccattcattccgttcctaactatggaacagggctgacaccttatatcctgtcaatgggcttgtatgtcggcggcatcatgctgacggttgtcttcccgctgaaggaagcgtcaggacgtccgagaaacggttttgagtggttcttcagcaaattcaatgtcatgatgcttgtaggaatcattcaatcattgattgtggcgacggttcttctattgggaattggtcttgaggtagagagcacgtggagattttacgtgtttacgatcattacgagtctcgctttcctggcgataattcaatttttggctacaacgatgggcaacccggggcgatttatcgcagtcattattttagtgctgcagctgggagcgagcggaggaaccttcccgctggaattgcttccgaatttctatcaggtcatccacggcgcattgccgatgacctacagcattaacggattcagagcagtcatttcaaacggtgactttggctacatgtggcagatggccggcgtcttaatcggcattgccctcgtcatgattgcactaagcattacttactttacaatgttaagccggaaggaagaaacatcagaagaacagccggcttcataaCGAG +DS_yhgE,pOpen_v3,dna,GTCTagaaaaagaacttgtttccttggaaataagttctttttttgttacatcccccatttaataagcaatcctgtataggttaatccgccgccaaacccgaaaagcaaaacgatttgatcttttttcagcttcccggctttcactgcgagatcgagcgccaaaacaattgaaaccgaagacgtgtttccgtagtgctcaacactagtgagcgttttttcaatcgggaacggtgttttttcacaaattgactcgatcatgcgcaagttggcgctgtgaggaacaaaccaatcgagatcatcggagctgagtcctgctttatgtaaaagccgttcaaattcgccagggacggttcttgcggcccatttatatacctcgcgtccgttttgcaccatttttccggaaccgacaagctgcacaccgtttatttcatttcgcagtccggcacgatacaaaatatcgccgccgttcccgcttgttccttgtacagacgcaagaaatcccggcgtctcttcatctcgttctactaacagcgcacccgcggcatcgccgaacagtacgcatgtcgttcgatcggtataatcggttacctttgataacgtctctccggcgatgacgagaattttttgatgaaggccagatgtgatcaatccatttgccaaatggaggccgtatgtcagcccggcgcatgtcgcattaatatccagcgcgccggtgctttcccagccgaaatattcctgtacgcggcatgccgtactcggaaaggcgtaatcggatgtggttgtgAAGC 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+pMTLB,pOpen_v3,dna,GTCTaaattcatattacttagaggatactattgagagagctattgataagatggttgagacattacctgagagccaaaaaactttttatgaatatgaattaaaaaaaagaaccaacaaaggctgagacagactccaaacgagtctgtttttttaaaaaaaatattaggagcattgaatatatattagagaattaagaaagacatgggaataaaaatattttaaatccagtaaaaatatgataagattatttcagaatatgaagaactctgtttgtttttgatgaaaaaacaaacaaaaaaaatccacctaacggaatctcaatttaactaacagcggccaaactgagaagttaaatttgagaaggggaaaaggcggatttatacttgtatttaactatctccattttaacattttattaaaccccatacaagtgaaaatcctcttttacactgttcctttaggtgatcgcggagggacattatgagtgaagtaaacctaaaaggaaatacagatgaattagtgtattatcgacagcaaaccactggaaataaaatcgccaggaagagaatcaaaaaagggaaagaagaagtttattatgttgctgaaacggaagagaagatatggacagaagaacaaataaaaaacttttctttagacaaatttggtacgcatataccttacatagaaggtcattatacaatcttaaataattacttctttgatttttggggctattttttaggtgctgaaggaattgcgctctatgctcacctaactcgttatgcatacggcagcaaagacttttgctttcctagtctacaaacaatcgctaaaaaaatggacaagactcctgttacagttagaggctacttgaaactgcttgaaaggtacggttttatttggaaggtaaacgtccgtaataaaaccaaggataacacagaggaatccccgatttttaagattagacgtaaggttcctttgctttcagaagaacttttaaatggaaaccctaatattgaaattccagatgacgaggaagcacatgtaaagaaggctttaaaaaaggaaaaagagggtctgccaaaggttttgaaaaaagagcacgatgaatttgttaaaaaaatgatggatgagtcagaaacaattaatattccagaggccttacaatatgacacaatgtatgaagatatactcagtaaaggagaaattcgaaaagaaatcaaaaaacaaatacctaatcctacaacatcttttgagagtatatcaatgacaactgaagaggaaaaagtcgacagtactttaaaaagcgaaatgcaaaatcgtgtctctaagccttcttttgatacctggtttaaaaacactaagatcaaaattgaaaataaaaattgtttattacttgtaccgagtgaatttgcatttgaatggattaagaaaagatatttagaaacaattaaaacagtccttgaagaagctggatatgttttcgaaaaaatcgaactaagaaaagtgcaataaactgctgaagtatttcagcagttttttttatttagaaatagtgaaaaaaatataatcagggaggtatcaatatttaatgagtactgatttaaatttatttagactggaattaataattaacacgtagactaattaaaatttaatgagggataaagaggatacaaaaatattaatttcaatccctattaaattttaacaagggggggattaaaatttaattagaggtttatccacaagaaaagaccctaataaaatttttactagggttataacactgattaatttcttaatgggggagggattaaaatttaatgacaaagaaaacaatcttttaagaaaagcttttaaaagataataataaaaagagctttgcgattaagcaaaactctttactttttcattgacattatcaaattcatcgAAGC +pNG168,pOpen_v3,dna,GTCTaccatagagttgagtagaatatttgacgaatgggaaacacatcatcgaaatttggtggttctacactgtgcgggatcgtcagacaccgactgttacctttaagttgcatccgtgaaaacaaggggccaaggcaggacagagtaatcactggtcggcgttggaagcctgaagaaggccccggacacgaacctctcgtgtccccacaggttgctacctcggccttcttccgctgtgggcttaagcgttaccccacgtagccgctgcgctgccaaatgaaatgtcctcatcacgatgacagcgatgcaaactacacagacacaccaagaccgaggtgaccgcgagtagctgacgctactccagaccaatgactcacaggtacagactcaaacgacaatccgggtccgatgctgccaggcttcgccccggatgtcgttgtgaagcatcgtcgcagcggaccagcagagccggtcctcaacacggatggagcggtattatcagtacgctcctatctacaatgtggacgcgaggggtcaaattggtttcgggaccctcgcatttgccggcttccacgcagtccgcacttctccacggactgcatgcacacgcctaatcaacaacaaggaatccggaagatcgttccaggagggaccctctctactgctggaattaccattaccgaggtcaccccacgggtaacagaatggatacctgaccttcttgaggaactactgccacggagtattcaatcggttcgaaaattcatccggcaagaggacccagaagtcctcacgcacgcacgatacaacaccgtctatcgccgacttcaagaggaaaccctgaggttcgaccatcaagaatggtgctccacaacggatatttggagtgatgcagaggctgaagcggttgaatacgtagaatcacttgtcgagttcgcagtcaaatattctgacgttgacgaagatgatctcgacgaactttcggagtatcaccagcagcggtgcaaatcgcttaaacagactctcactacgatcagtaccggacgtggcccactcaacgctggccttgaagccctcgccaagggacccgtacgactccacgatgagctcgatgacgcaccgcaaccgatcacgcttgtccttgatggcgagttgtggtcaaaactcgacgatagaggaacaggtatccgagcacttgcagctatcgccgtgctcggctccaccttcgacgtccgcctggttatttcaccagcgttagacgccgcgattgagcgacggtatccagactggtatgactcccatctccgtcttactgaaacccgtgaaacctcctctgtagaatcagcgggtggcgacggacaaccatcggcggagcagctcgaggaagcatgggaagcaatacagaacctcccagaggaatcagggagactccgacttctccggaacctccctatagagggctctcgagactatcgtgatctcaagcaggatgatgagattgacgtgcaggcagggactgtcggccgttacatcctcgatctagaagaattagggcttgtggatatcgaccgtcgtggacaatacaacagcgcatccctcaccggcttaggacaagtagcagttgagcagtatgtcaccacggactaccgggtgatccatccgacccaatcgacgctggaaacgcatcttacgccgacccctcagccccaagcaagtacagtgtatcccgcgcgatcggacacgagggagggggatcagcctgggacagcggaggattggatagctgcgacaggcagtcctagtgagggtgctgactacgttcaatggctcgatgggccgtctggtgttctcgacgcttggggaatgcatcagcggtaccttgctggccgtcgagatcgtggtgtcaccctagtcgatgaccgtatcgagcgtttcgaggacggtcgcgtatcgtatctcagttgcttcgacgatgatctcttcgtagccactcagtggggcggcccccttcctaccctgggacgtattgcgggtgccttactcagcgacaaagcactgagtaagatccttaccccttctcgactcggaaaccagttcgaagaaatcaacgatgcggtcgttgaacaactcgatagagaagctggggaaatcatccgtagaggacaccaaatcggctggttcagcgaggacgaagaggactacgatggttggcgtgagcgtattggctccgttcgcagtctgtgtttgcaacaagttggtgaactcacaaacagtgacgacgtcgaagcccggactgagttactacgtgacctgcacggcttggttgcctcagcgacacagctgtactacgcagccggtgtagacgtcacaatcaacgttcgagtccctgacacgggaatgctcatctcggatgaacgccgactcgacgatttcctcggtttcgcccggtacaccattccgaaacagtccgtgtacgggattcattcggggtataggatgctcctcgaggatcgaccggagaaactcaaacgacgacttccctacgaggtggatgatgctgattccacaatgcatctgaccgcatcgtgggttttctccggatcaacgatgatcgatcttcacgatgacatcgaggacgccattgaaatggaaaccaacgaaattcgtgaagcaattgctaatggacaggagtcagcaccggtaatggaaatccccgtccagattgggaactcctactcagcaattcgtaatcacgtcgaggactacgcttcagcgaagaactaccaggtagctcaccaggaggatattcacgaaggaaaacaggatctcgagcgactcgtccgcctgttcctccgtgtcctcgggacagaggatcgaccacatcgagcgtgtcctcacgacgttgccgaggcaatgcttcatgttgctcaatcctcccggaactacgatttcatcaccgtgcgagacatctcgtacggactgtcgaatctccccacgaaacgactcttacccgagctcccaccaacagcaacgaagctcctcaaaaccctgcttgatgcggatgacccgatgggacggtctgaaatcatcgataccgctgacatttcggaaagtagctatgatcgctacatcaacgaactcgccgcttgggatatcattgaaccccgcgaaatcgagggacaccgtcggtgggaagctcacttagagccgtggtggacaccgcagagtgatcgtgacgaaccctatgccgaccctgaccccgacacgggaatactgtacgctgaatttccccgtgatgtcgctagtgcggtgatgtgccacctcatcacccactacgacttacccgaccttgagacagcgtatcttgagggtatccaaccgggggacgacatcaaggccctcttcgatgatcacgaccgactcagacgatggcggccattcctctggggtgcattcgccgactcggacaaacttgaaagaggcccatctggtacagcagcttcggactcgaccgtagttcgactcggtcaatctccaggacccgacacagcacaatcgagcttccaagatgtctcagaaacagcaacccaacgagatcgactcagtcaaccgtcgccagggctggactaatgattgacggcaatcgataactacgcaatgaatcctccacaaccagcatctgaaccgtactcgaaaggtgatgcagtaaccgtatacgtgggcAAGC +pSK41,pOpen_v3,dna,GTCTgatttatcggcctcataattttagatatattaattatattagagatttgttatcaaatacatttcattacaagcatataattgacctgtgaggcaacaaaaaatttttaatttttttaaaatagtgttactattgaacatagaaaaagacacctaacgcttggcggttgttagatgtctaaagagtttaaataatatgtgttatttacttttacaatttgtttgtatctttattatatatacaacctcgatacattacaagataaaagcaacaagtctttaagttatatataacggataacattatataaccttattcatcacttgtaaataacgatattacttaaactccttaaatatacatttaaaggagtttttatcatgtctaaacaattttttacagtagaagaaaattataaagaacgcttttaccaattacctaaagtattttttactaatcctaattataaagatttatcgaatgacgctaaaattgcttacgcaatattgagagatcgtttacaattatctattaaaaataactggatagatacagaaggtaacatatatttcatttatacagttgctgatttagaagtgatattaaattgtgggaacaaaaaaatcactaaaattaaaaaagaattagaaaatgtagatttactaatccaaaaaaggcagggtttaaacaaaccaaatctactttatttattaaaaccagccataactaaaaatgatatttatgaaatagataaggctgaaaatgaagttgaggccttacaagacaaggaagtgtcaaaaggacacgtccagaaatgtcaaaaggacacgtctagaaatgtcaaaaggacacgtctagaaatgtcaaaaggacacacaaatgatactgattttattgatactgattttattgatactgaaagtaatgatatgaataatatgaatgatacaaatcaacattcaaatcattcaaatcacttttcaaatattcatgataaagaatctcttaaatatatagaacttcaagaactccccgaattaattaaaagttatattaataatttcagctatgaagaagttaaaagtattaaatctgttattttaaaagctaaaaaatcatttaataacaaatacgatactttctatatgttagaagatatagatgaagaattactattggttttaaaacgatttaaaggttatcttgttaaaaaacaagaaaaagtagctaatatggaaggttatttaatgagaagtatcattgctgaacttgaagaaatgcactcaactattatgagaagaaaaaatatggaaaacaatccattatctctatttaattagtttttgaagttaaattttcaaaaaaaagagattatacacctaaaaaggtgtataatctttacAAGC From acd86a87de8b4713bd405a16787ceef474f6d341 Mon Sep 17 00:00:00 2001 From: Keoni Gandall Date: Sun, 20 Oct 2024 22:26:05 -0700 Subject: [PATCH 7/9] update --- parts/linkers.md | 17 +++--- parts/parts/bsub.yaml | 104 ++++++++++++++++----------------- parts/parts/meganucleases.yaml | 40 +++++++++++++ parts/parts/openenzyme.yaml | 30 ++++++++++ parts/parts/proteins/HSlig4.md | 6 ++ 5 files changed, 136 insertions(+), 61 deletions(-) create mode 100644 parts/parts/meganucleases.yaml create mode 100644 parts/parts/openenzyme.yaml create mode 100644 parts/parts/proteins/HSlig4.md diff --git a/parts/linkers.md b/parts/linkers.md index 1bb5f8b..47eae6e 100644 --- a/parts/linkers.md +++ b/parts/linkers.md @@ -30,15 +30,14 @@ overhangs with numbers: 14. ACGA 15. TATC 16. TAGG -17. TACA -18. TTAC -19. TTGA -20. TGGA -21. GAAG -22. GACC -23. GCCG -24. AAAA -25. GGGG +17. TTAC +18. TTGA +19. TGGA +20. GAAG +21. GACC +22. GCCG +23. AAAA +24. GGGG Once a plasmid cloned using a linker, its new definition is derived from the particular linkers used to construct it. For example, an assembly with A1(2) + B1(4) creates a new plasmid with a fragment defined with the overhangs 2 and 4. diff --git a/parts/parts/bsub.yaml b/parts/parts/bsub.yaml index 291dc0d..576eadc 100644 --- a/parts/parts/bsub.yaml +++ b/parts/parts/bsub.yaml @@ -42,14 +42,14 @@ US_amyE: sequence: ttaacaaaattctccagtcttcacatcggtttgaaaggaggaagcggaagaatgaagtaagagggatttttgactccgaagtaagtcttcaaaaaatcaaataaggagtgtcaagaatgtttgcaaaacgattcaaaacctctttactgccgttattcgctggatttttattgctgtttcatttggttctggcaggaccggcggctgcgagtgctgaaacggcgaacaaatcgaatgagcttacagcaccgtcgatcaaaagcggaaccattcttcatgcatggaattggtcgttcaatacgttaaaacacaatatgaaggatattcatgatgcaggatatacagccattcagacatctccgattaaccaagtaaaggaagggaatcaaggagataaaagcatgtcgaactggtactggctgtatcagccgacatcgtatcaaattggcaaccgttacttaggtactgaacaagaatttaaagaaatgtgtgcagccgctgaagaatatggcataaaggtcattgttgacgcggtcatcaatcataccaccagtgattatgccgcgatttccaatgaggttaagagtattccaaactggacacatggaaacacacaaattaaaaactggtctgatc DS_amyE: description: downstream flank amyE - prefix: GTCT - suffix: AAGC + prefix: AAGC + suffix: ATAG tags: [downstream_flank] sequence: agagtcgacatggatgagcgatgatgatatccgtttaggctgggcggtgatagcttctcgttcaggcagtacgcctcttttcttttccagacctgagggaggcggaaatggtgtgaggttcccggggaaaagccaaataggcgatcgcgggagtgctttatttgaagatcaggctatcactgcggtcaatagatttcacaatgtgatggctggacagcctgaggaactctcgaacccgaatggaaacaaccagatatttatgaatcagcgcggctcacatggcgttgtgctggcaaatgcaggttcatcctctgtctctatcaatacggcaacaaaattgcctgatggcaggtatgacaataaagctggagcgggttcatttcaagtgaacgatggtaaactgacaggcacgatcaatgccaggtctgtagctgtgctttatcctgatgatattgcaaaagcgcctcatgttttccttgagaattacaaaacaggtgtaacacattctttcaatgatcaactgacgattaccttgcgtgcagatgcgaatacaacaaaagccgtttatcaaatcaataatggaccagagacggcgtttaaggatggagatcaattcacaatcggaaaaggagatccatttggcaaaacatacaccatcatgttaaaaggaacgaacagtgatggtgtaacgaggaccgagaaatacagttttgttaaaagagatccagcgtcggccaaaaccatcggctatcaaaatccgaatcattggagccaggtaaatgcttatatctataaacatgatgggagccgagtaattgaattgaccggatcttggcctggaaaaccaatgactaaaaatgcagacggaatttacacgctgacgctgcctgcggacacggatacaaccaacgcaaaagtgatttttaataatggcagcgcccaagtgcccggtcagaatcagcctggctttgattacgtgctaaatggtttatataatgactcgggcttaagcggttctcttccccattgagggcaaggctagacgggacttaccgaaagaaaccatcaatgatggtttcttttttgttcataaatcagacaaaacttttctcttgcaaaagtttgtgaagtgttgcacaatataaatgtgaaatacttcacaaacaaaaagacatcaaagagaaacataccctg camR(Bsub): description: Chloramphenicol resistance in Bacillus subtilis - prefix: AAGC - suffix: ATAG + prefix: GTCT + suffix: AAGC tags: [selection] sequence: tccggcaaaaaagggcaaggtgtcaattctcatgtttgacagcttatcatcggcaatagttacccttattatcaagataagaaagaaaaggatttttcgctacgctcaaatcctttaaaaaaacacaaaagaccacattttttaatgtggtctttattcttcaactaaagcacccattagttcaacaaacgaaaattggataaagtgggatatttttaaaatatatatttatgttacagtaatattgacttttaaaaaaggattgattctaatgaagaaagcagacaagtaagcctcctaaattcactttagataaaaatttaggaggcatatcaaatgaactttaataaaattgatttagacaattggaagagaaaagagatatttaatcattatttgaaccaacaaacgacttttagtataaccacagaaattgatattagtgttttataccgaaacataaaacaagaaggatataaattttaccctgcatttattttcttagtgacaagggtgataaactcaaatacagcttttagaactggttacaatagcgacggagagttaggttattgggataagttagagccactttatacaatttttgatggtgtatctaaaacattctctggtatttggactcctgtaaagaatgacttcaaagagttttatgatttatacctttctgatgtagagaaatataatggttcggggaaattgtttcccaaaacacctatacctgaaaatgctttttctctttctattattccatggacttcatttactgggtttaacttaaatatcaataataatagtaattaccttctacccattattacagcaggaaaattcattaataaaggtaattcaatatatttaccgctatctttacaggtacatcattctgtttgtgatggttatcatgcaggattgtttatgaactctattcaggaattgtcagataggcctaatgactggcttttataatatgagataatgccgactgtactttttacagtcggttttctaatgtcactaacctgccccgttagttgaagaaggtttttatattacagctccagatcctctacgccggacgcatcgtggccggcatcaccggcgccacaggtgcggttgctggcgcctatatcgccgacatcaccgatggggaagatcgggctcgccacttcgggctcatgagcgcttgtttcggcgtgggtatggtggcaggccccgtggccgggggactgttgggcgccatctccttgcatgcaccattccttgcggcggcggtgctcaacggcctcaacctactactgggctgcttcctaatgcaggagtcgcataagggag US_lacA: @@ -60,14 +60,14 @@ US_lacA: sequence: gtgatgtcaaagcttgaaaaaacgcacgtaacaaaagcaaaatttatgctccatgggggagactacaaccccgatcagtggctggatcggcccgatattttagctgacgatatcaaactgatgaagctttctcatacgaatacgttttctgtcggcatttttgcatggagcgcacttgagccggaggagggcgtatatcaatttgaatggctggatgatatttttgagcggattcacagtataggcggccgggtcatattagcaacgccgagcggagcccgtccggcctggctgtcgcaaacctatccggaagttttgcgcgtcaatgcctcccgcgtcaaacagctgcacggcggaaggcacaaccactgcctcacatctaaagtctaccgagaaaaaacacggcacatcaaccgcttattagcagaacgatacggacatcacccggcgctgttaatgtggcacatttcaaacgaatacgggggagattgccactgtga DS_lacA: description: downstream flank lacA - prefix: GTCT - suffix: AAGC + prefix: AAGC + suffix: ATAG tags: [downstream_flank] sequence: ttcaagctatatttggagttgagcctcttgaaacggacaccctgtatccgaaggatcgaaacgctgtcagctaccgcagccaaatatatgaaatgaaggattatgcaaccgtgattgatgtaaagacagcttcagtggaagcggtgtatcaagaagatttttatgcgcgcacgccagcggtcacaagccatgagtatcagcagggcaaggcgtattttatcggcgcgcgtttggaggatcaatttcagcgtgatttctatgagggtctgatcacagacctgtctctctctccagtttttccggttcggcacggaaaaggcgtctccgtacaagcgaggcaggatcaggacaatgattatatttttgtcatgaatttcacggaagaaaaacagctggtcacgtttgatcagagtgtgaaggacataatgacaggagacatattgtcaggcgacctgacgatggaaaagtatgaagtgagaattgtcgtaaacacacattag eryR(Bsub): description: erythromycin resistance in Bacillus subtilis - prefix: AAGC - suffix: ATAG + prefix: GTCT + suffix: AAGC tags: [selection] sequence: GAGTTAACAAGAGTTTGTAGAAACGCAAAAAGGCCATCCGTCAGGATGGCCTTCTGCTTAGCTAGAGCGGCGGATTTGTCCTACTCAGGAGAGCGTTCACCGACAAACAACAGATAAAACGAAAGGCCCAGTCTTTCGACTGAGCCTTTCGTTTTATTTGATGCCTCAAGCTAGAGAGTCCTAGAGTCTAGGGACCTCTTTAGCTCCTTGGAAGCTGTCAGTAGTATACCTAATAATTTATCTACATTCCCTTTAGTAACGTGTAACTTTCCAAATTTACAAAAGCGACTCATAGAATTATTTCCTCCCGTTAAATAATAGATAACTATTAAAAATAGACAATACTTGCTCATAAGTAACGGTACTTAAATTGTTTACTTTGGCGTGTTTCATTGCTTGATGAAACTGATTTTTAGTAAACAGTTGACGATATTCTCGATTGACCCATTTTGAAACAAAGTACGTATATAGCTTCCAATATTTATCTGGAACATCTGTGGTATGGCGGGTAAGTTTTATTAAGACACTGTTTACTTTTGGTTTAGGATGAAAGCATTCCGCTGGCAGCTTAAGCAATTGCTGAATCGAGACTTGAGTGTGCAAGAGCAACCCTAGTGTTCGGTGAATATCCAAGGTACGCTTGTAGAATCCTTCTTCAACAATCAGATAGATGTCAGACGCATGGCTTTCAAAAACCACTTTTTTAATAATTTGTGTGCTTAAATGGTAAGGAATACTCCCAACAATTTTATACCTCTGTTTGTTAGGGAATTGAAACTGTAGAATATCTTGGTGAATTAAAGTGACACGAGTATTCAGTTTTAATTTTTCTGACGATAAGTTGAATAGATGACTGTCTAATTCAATAGACGTTACCTGTTTACTTATTTTAGCCAGTTTCGTCGTTAAATGCCCTTTACCTGTTCCAATTTCGTAAACGGTATCGGTTTCTTTTAAATTCAATTGTTTTATTATTTGGTTGAGTACTTTTTCACTCGTTAAAAAGTTTTGAGAATATTTTATATTTTTGTTCATGTAATCACTCCTTCTTAATTACAAATTTTTAGCATCTAATTTAACTTCAATTCCTATTATACAAAATTTTAAGATACTGCACTATCAACACACTCTTAAGTTTGCTTCTAAGTCTTATTTCCATAACTTCTTTTACGTTTCCGCCATTCTTTGCTGTTTCGATTTTTATGATATGGTGCAAGTCAGCACGAACACGAACCGTCTTATCTCCCATTATATCTTTTTTTGCACTGATTGGTGTATCATTTCGTTTTTCTTTTGTGCTAGAGGATCAATTCTTGAAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTG US_thrC: @@ -78,8 +78,8 @@ US_thrC: sequence: aattcatgtaaaagatgaggttggttcattctcgaaaattacatctgtgttctcagagcggggcgtgagctttgaaaaaatccttcagctgccaattaaaggccatgatgagttagctgaaatcgtaattgtcacacatcatacatcagaagctgatttcagtgatatcctgcaaaacctaaatgatttggaagtcgttcaagaagtcaaaagcacatatcgtgtagaagggaacggttggagctaatgtggaaaggacttatccatcaatataaagaatttttacctgtaacagatcaaacaccggcgctaactttacatgaaggaaacacacctcttattcacctgccgaagctgtctgagcagctcggaattgagcttcatgtcaaaacggaaggcgtcaatcctacgggatcatttaaagatcgcggaatggttatggctgtggcaaaggcaaaagaagaaggcaatgacacgattatgtgcgcgtcaacaggtaacacttccgctgctgcggcagcatatgcagcccgtgctaacatgaaatgcattgtcatcatcccgaacggaaaaattgcatttggaaaactcgctcaagctgtcatgtacggagccgagattatcgcaattgacggaaactttgacgatgcgcttaaaattgtccgttccatctgtgagaaatcaccgattgcccttgtcaactcagtcaacccttaccgcattgaaggccaaaaaactgctgccttcg DS_thrC: description: - prefix: GTCT - suffix: AAGC + prefix: AAGC + suffix: ATAG tags: [downstream_flank] sequence: AGGCCTTCCGAAAATGCGCGGCTTTGAAGCTGAGGGTGCGGCGGCAATCGTGCGCAATGAAGTGATTGAAAATCCGGAAACAATAGCGACAGCCATTCGTATCGGAAACCCGGCAAGCTGGGACAAAGCTGTAAAGGCAGCCGAGGAATCCAATGGGAAAATTGACGAAGTCACTGATGATGAAATCCTTCACGCATATCAGCTGATCGCCCGTGTAGAAGGCGTGTTTGCAGAACCAGGTTCTTGCGCGTCTATCGCAGGAGTGCTGAAACAGGTGAAATCCGGAGAAATTCCGAAAGGCAGCAAGGTCGTAGCTGTGTTAACAGGAAACGGACTGAAAGATCCGAACACAGCGGTCGACATTTCAGAAATCAAGCCTGTCACATTGCCGACTGATGAAGACAGCATCCTTGAATATGTAAAAGGAGCGGCCCGTGTATGAACGAAGCCGACATGCTGTTCTCTGTCACTGTTCCCGGAAGCACAGCTAACCTAGGCCCCGGCTTTGATTCAGTCGGAATGGCGCTCAGCAGATATTTGAAGCTGACCGTCTTTGAAAGCGACAAATGGTCTTTTGAGGCTGAAACAGAAACAGTCGCCGGAATT Pars: @@ -108,8 +108,8 @@ US_glmS: sequence: CAACATGCCGCTTCTGTCTAAGAAACCGCTCTTCATTTTCCTTTCTCAAAGCGGAGAAACAGCAGACAGCCGCGCGGTACTCGTTCAAGTCAAAGCGCTCGGACACAAAGCCCTGACAATCACAAACGTACCTGGATCAACGCTTTCTCGTGAAGCTGACTATACATTGCTGCTTCATGCAGGCCCTGAGATCGCTGTTGCGTCAACGAAAGCATACACTGCACAAATCGCAGTTCTGGCGGTTCTTGCTTCTGTGGCTGCTGACAAAAATGGCATCAATATCGGATTTGACCTCGTCAAAGAACTCGGTATCGCTGCAAACGCAATGGAAGCTCTATGCGACCAGAAAGACGAAATGGAAATGATCGCTCGTGAATACCTGACTGTATCCAGAAATGCTTTCTTCATCGGACGCGGCCTTGACTACTTCGTATGTGTCGAAGGCGCACTGAAGCTGAAAGAGATTTCTTACATCCAGGCAGAAGGTTTTGCCGGCGGTGAGCTAAAGCACGGAACGATTGCCTTGATCGAACAAGGAACACCAGTATTCGCACTGGCAACTCAAGAGCATGTAAACCTAAGCATCCGCGGAAACGTCAAAGAAGTTGCTGCTCGCGGAGCAAACACATGCATCATCTCACTGAAAGGCCTAGACGATGCGGATGACAGATTCGTATTGCCGGAAGTAAACCCAGCGCTTGCTCCGTTGGTATCTGTTGTTCCATTGCAGCTGATCGCTTACTATGCTGCACTGCATCGCGGCTGTGATGTGGATAAACCTCGTAACCTTGCGAAGAGTGTTACTGTGGAGTAATAAATGTTTAACCCCTTTGGATAAGATTATCTAAAGGGGTGT DS_glmS: description: downstream glmS flank - prefix: GTCT - suffix: AAGC + prefix: AAGC + suffix: ATAG tags: [downstream_flank] sequence: TTATGTCCAAAAGCGAAAGAACGTCTACAGTGGAAAAATGGATTAAAGAAGTTGATGGTACAGGACCCGACTATCGATCGACCATGGCTATAGATTTAAGATGTTTCCTCATTAGATCGGTCAATCAGGTTAAAAGGCATTAAAACTTGGAGAGAAACTGCTTTTGCCTAACTGAGTTTTCTAATGTTAATACAGATATAAGAGAGCAGTTCCTCATTAAGATACGCTTTTGATAGCAGTTGGAATTTAGCATGCTAGAAATCCGAAAACCGGAGAGCCTATTGGAATGATAACGTGTTTTTTTATTGACAATTAATAAAGGGCAAAGAATTCTAGAAGTCGCACTTACAATGTACTTGTGGGATTAGGTTTTAAAGTTCATTCTTTTTTTATTGAGTACATTCCGGTTAAAGTTGAAATGATTCAGTAAAGGACACGCCATAAATAAAAGGTAGCGTATGATTGCCATTCTTTCCATGGAACCGATATTTCTAAGATTTCATCTTTCGTAGGCTTTCGGTTCATATTACGTAATATTTTTATCGAATGAATAAGACCCACATCATCGATTGGAAAAGCTGTCGGGAACCTAAGGCAGCGCATTAGAACATAATTGGCTGTCCAAGGACCGATTCCTCGTATTTTAATTAAGTTTTTTTCAGCATCTTTAAAGTTCATTTTCATTAATTTTTCCCTCGATAATTCTCCACTTGCCATTAATCTGGCTATTCCAATGATATATTCACTTTTTTTCACCGTCATTTTAATATCTGCCAGGTCGGTAGGGGTTAACCGTGCAATTCGCTCGTACGGTGGGAACACCCAATACTTTTTACCATTCCATTCAATAGAATCGCCAAATGCTTCTACAAATTGCTTCTTTAAGGAGTACGCGAAGGCTAA Pmtl-short: @@ -144,14 +144,14 @@ US_sigF: sequence: gctgaacttggaggacctttcctttatggacagctcggggcttggcgttattttaggaagatacaagcaaattaagcaaattggcggagaaatggttgtttgcgctatctctcctgcggtgaagcgattgtttgatatgtcgggtctgtttaaaattatccgatttgaacaatctgaacagcaggcactcctgacactgggggtggcatcatgaaaaatgaaatgcaccttgagttttctgccctcagtcagaatgaatcgttcgcccgtgtgacagttgcttcatttatagctcagctggacccgacaatggatgaactgactgaaatcaaaacagtcgtgtcagaggctgtcacgaatgcgattatccatggatatgaagagaactgtgaagggaaagtttacatttcagtgacgctggaagatcatgtcgtatatatgactattcgtgatgaaggcttaggcattacagatcttgaagaagcccgtcagcctctatttacgactaagcctgagcttgagcgctctggaatgggctttaccattatggaaaatttcatggatgatgtcagtatcgattcatcgcctgaaatgggaacaacgattcgcttaacaaagcacttatcaaaaagcaaagcgctttgtaattaaggagatttgttatggatta DS_sigF: description: downstream flank sigF - prefix: GTCT - suffix: AAGC + prefix: AAGC + suffix: ATAG tags: [downstream_flank] sequence: tctgcagtgcaggctagcttttttgtgcaaaagcgtggtaatttatggtcttttcgagcggatgaatgagaacaaaatcgaaccacatactacatatataaccaccgaaagatggtgatcaatgatggaacgacgaatatttatccggcttcgccaccgagtgctggcacatccaggggatattattaccgttggagatgccgcgcaaatagaagggcagcttcagctgaaaaagaaactttcggctatgccgctttatcaggtgagcgaaaaagataaaaatatcgtaattctggatatcatacaagtcctcagagccattcatttacaagacccgacaattgatgttcaaaccgtaggcggagcagaaaccattgttgaaattcagtatcgaaagcgaaatttatcaacggttctatttatcggtgtctggctgcttctgtttattggatcgtgtcttgccatcatgaactttcatgaggatgtaagcatgagagatgttcatatcgcactatatgaaatcataaccggagagaggaatgactatccatatttgcttcaaatcccatacagcatcggtttgggactggggatgatcgtgttttttaaccacatatttaaaaagcgcctaaatgaagagcccagcccgctggaggttgagatgtttaactatcagcttgatctcgatcaatatgtggccatgcatgagaatcaagaaaccataaaggatctgcatgatcgttagtgtattgttcatcatttttgtcgggctcggcggaggcataacggtgggggcgggtttcgttgcttttttaaccgtaatgggaatcattccgcggctgatgcagctcaccaaaacaatgagatttgttcaggcttatgaagcg kanR(Bsub): description: bacillus subtilis kanamycin resistance - prefix: AAGC - suffix: ATAG + prefix: GTCT + suffix: AAGC tags: [selection] sequence: gaggtcatcgttcaaaatggtatgcgttttgacacatccactatatatccgtgtcgttctgtccactcctgaatcccattccagaaattctctagcgattccagaagtttctcagagtcggaaagttgaccagacattacgaactggcacagatggtcataacctgaaggaagatctgattgcttaactgcttcagttaagaccgaagcgctcgtcgtataacagatgcgatgatgcagaccaatcaacatggcacctgccattgctacctgcacagtcaaggatggtagaaatgttgtcggtccttgcacacgaatattacgccatttgcctgcatattcaaacagctcttctacgataagggcacaaatcgcatcgtggaacgtttgggcttctaccgatttagcagtttgatacactttctctaagtatccacctgaatcataaatcggcaaaatagagaaaaattgaccatgtgtaagcggccaatctgattccacctgagatgcataatctagtagaatctcttcgctatcaaaattcacttccaccttccactcaccggttgtccattcatggctgaactctgcttcctctgttgacatgacacacatcatctcaatatccgaatagggcccatcagtctgacgaccaagagagccataaacaccaatagccttaacatcatccccatatttatccaatattcgttccttaatttcatgaacaatcttcattctttcttctctagtcattattattggtccattcactattctcattcccctttcagataattttagatttgcttttctaaataagaatatttggagagcaccgttcttattcagctattaaacccattatatcgggtttttgaggggatttcaactgcag US_yhgE: @@ -162,14 +162,14 @@ US_yhgE: sequence: gacgggtctaaacaggtccaaagcggatcatcaaaacttgcagacggatctgcatcacttgatacaggtttaggcaagctattggacggcacaggagagctgtctagtaagctgaaggatgcggctgaccaaacaggagatatcgatgctgatgatcaaacatacggcatgtttgcggatccggttaaaaccaaagatgatgccattcattccgttcctaactatggaacagggctgacaccttatatcctgtcaatgggcttgtatgtcggcggcatcatgctgacggttgtcttcccgctgaaggaagcgtcaggacgtccgagaaacggttttgagtggttcttcagcaaattcaatgtcatgatgcttgtaggaatcattcaatcattgattgtggcgacggttcttctattgggaattggtcttgaggtagagagcacgtggagattttacgtgtttacgatcattacgagtctcgctttcctggcgataattcaatttttggctacaacgatgggcaacccggggcgatttatcgcagtcattattttagtgctgcagctgggagcgagcggaggaaccttcccgctggaattgcttccgaatttctatcaggtcatccacggcgcattgccgatgacctacagcattaacggattcagagcagtcatttcaaacggtgactttggctacatgtggcagatggccggcgtcttaatcggcattgccctcgtcatgattgcactaagcattacttactttacaatgttaagccggaaggaagaaacatcagaagaacagccggcttcataa DS_yhgE: description: downstream_flank yhgE - prefix: GTCT - suffix: AAGC + prefix: AAGC + suffix: ATAG tags: [downstream_flank] sequence: agaaaaagaacttgtttccttggaaataagttctttttttgttacatcccccatttaataagcaatcctgtataggttaatccgccgccaaacccgaaaagcaaaacgatttgatcttttttcagcttcccggctttcactgcgagatcgagcgccaaaacaattgaaaccgaagacgtgtttccgtagtgctcaacactagtgagcgttttttcaatcgggaacggtgttttttcacaaattgactcgatcatgcgcaagttggcgctgtgaggaacaaaccaatcgagatcatcggagctgagtcctgctttatgtaaaagccgttcaaattcgccagggacggttcttgcggcccatttatatacctcgcgtccgttttgcaccatttttccggaaccgacaagctgcacaccgtttatttcatttcgcagtccggcacgatacaaaatatcgccgccgttcccgcttgttccttgtacagacgcaagaaatcccggcgtctcttcatctcgttctactaacagcgcacccgcggcatcgccgaacagtacgcatgtcgttcgatcggtataatcggttacctttgataacgtctctccggcgatgacgagaattttttgatgaaggccagatgtgatcaatccatttgccaaatggaggccgtatgtcagcccggcgcatgtcgcattaatatccagcgcgccggtgctttcccagccgaaatattcctgtacgcggcatgccgtactcggaaaggcgtaatcggatgtggttgtg spcR(Bsub): description: bacillus subtilis spectinomycin resistance - prefix: AAGC - suffix: ATAG + prefix: GTCT + suffix: AAGC tags: [selection] sequence: acgtgtaggctggagctgcttccctgcaggagtcaggtcaatcatcagataactatggcggcacgtgcattaaccacggttgtatcccgtctaaagtactcgtctgtgcaacaggtgacatcacgaaacgatcctcatcctgtctcttgatccatggattacgcgttaacccgggcccgcggatgcatatgatcagatcttaaggcctaggtctagaggatcgatctgtataataaagaataattattaatctgtagacaaattgtgaaaggatgtacttaaacgctaacggtcagctttattgaacagtaatttaagtatatgtccaatctagggtaagtaaattgagtatcaatataaactttatatgaacataatcaacgaggtgaaatcatgagcaatttgattaacggaaaaataccaaatcaagcgattcaaacattaaaaatcgtaaaagatttatttggaagttcaatagttggagtatatctatttggttcagcagtaaatggtggtttacgcattaacagcgatgtagatgttctagtcgtcgtgaatcatagtttacctcaattaactcgaaaaaaactaacagaaagactaatgactatatcaggaaagattggaaatacggattctgttagaccacttgaagttacggttataaataggagtgaagttgtcccttggcaatatcctccaaaaagagaatttatatacggtgagtggctcaggggtgaatttgagaatggacaaattcaggaaccaagctatgatcctgatttggctattgttttagcacaagcaagaaagaatagtatttctctatttggtcctgattcttcaagtatacttgtctccgtacctttgacagatattcgaagagcaattaaggattctttgccagaactaattgaggggataaaaggtgatgagcgtaatgtaattttaaccctagctcgaatgtggcaaacagtgactactggtgaaattacctcgaaagatgtcgctgcagaatgggctatacctcttttacctaaagagcatgtaactttactggatatagctagaaaaggctatcggggagagtgtgatgataagtgggaaggactatattcaaaggtgaaagcactcgttaagtatatgaaaaattctatagaaacttctctcaattaggctaattttattgcaataacaggtgcttacttttctggagttctttagcaaatttttttattagctgaacttagtattagtggccatactcctccaatccaaagctatttagaaagattactatatcctcaaacaggcggtaacgcgttgtccttttccgctttaataaactatggaagtatgtacagtcttgcaatgttgagtgaacaaacttccataataaaatatgcaagctggtaccctaaggaggatattcatatgcaagcact US_yrbD: @@ -180,97 +180,97 @@ US_yrbD: sequence: gtctgaattgcctgctgtgatctcattaatattcaaaagcgccttcgcattggattccgcttttggcggcttgatcggtatggcgatttcctggggcgttaaacgtggcatttactcaaacgaagccggccagggaacaggcccgcatccggcagctgctgcagaggtttctcatccggtcaagcagggactggttcaggcattttctgtttatattgacaccttgtttgtatgttccgctacagctttcatgatcttgtttaccggcatgtacaatacccaggctgcagatggttcctttatcgttcaccagcttaaaggcgtagaagcaggacctggtttcacacaagcggcaattgacagtgttcttcccggattcggtgcaggctttgttgccatcgctttattcttcttcgcttttacaacaattatggcgtattactacatcgcagaaacaaatatcgcctatttagcacgaggcagagaaagcaaatgggcgatgcttggcttaaaactgattattttagccgccacgttctatggtactgtgaaaacagcttcacttgcttgggcattaggggatgcagggcttggcattatggtatggctgaacgtcattgccattgtgctgcttgccaagccggcgctccttgctttaaaggattatgagcgccaaaagaagcagggcttagacccgatctttgatccaaaagcgctgggcatcaaaaacgctgatttctgggagaaagaatacactcatgaaagtgaacgtgtctcctgaacatactaaaaccggcccgatatgacctcgtgccggtttt DS_yrbD: description: downstream flank yrbD - prefix: GTCT - suffix: AAGC + prefix: AAGC + suffix: ATAG tags: [downstream_flank] sequence: ttatgaacgatagtttgtatacacttcttgaacatcatcgtcatcttcaaggacgtctattaatgtttccagcttctcaacagcctgatcatcggcttctgcataggtattgggaagcatggtcacctccgccgaagaaattagatatttcgattcgagggctgttttcacctcttcaaaatgctcaggctctgtgaaaacctcatataggtcttcttcaattcgcagctcttctcctcctgcttcaagcacatcaagcatgagctcatcttcttctatttgccgttcagtacggtctatcgtaatgaacccctttcgttcaaataaaaaggctacacatccgctttctcccagacttccgccatttttattaaaggctgtccgcacatttgacgccgttcggtttttattatctgttacgcattcaaccatgacggcaatccctgaagggccgtagccttcataggtgatttcttcatagctgcttccgtcttggcctcccgcagcttttttaatggcccgatctatattttcattcggcatattggcgccttttgctttttcgatgacaagacgcaatgctgaatttgcttctggatccgctccgccttctttcgctgccacatagatttcttttgccagcttcataaatattttcccgcgctttgcatcctgtgcattttttcttttttgaatgtttttccacttggaatggcctgccatgttttcaccttcttttacattttctatgtacactttaatgggaaatgttcgctgatgcaaagaaaaaggctgc TO_pE194ts: description: pE194ts is a temperature sensitive origin in Bacillus subtilis. Can be used for curable plasmids. - prefix: GTCT - suffix: AAGC + prefix: AAGC + suffix: ATAG tags: [origin] sequence: gacacctaaattcaaaatctatcggtcagatttataccgatttgattttatatattcttgaataacatacgccgagttatcacataaaagcgggaaccaatcatcaaatttaaacttcattgcataatccattaaactcttaaattctacgattccttgttcatcaataaactcaatcatttctttaattaatttatatctatctgttgttgttttctttaataattcatcaacatctacaccgccataaactatcatatcttctttttgatatttaaatttattaggatcgtccatgtgaagcatatatctcacaagacctttcacacttcctgcaatctgcggaatagtcgcattcaattcttctgtaattatttttatctgttcataagatttattaccctcatacatcactagaatatgataatgctcttttttcatcctatcttctgtatcagtatccctatcatgtaatggagacactacaaattgaatgtgtaactcttttaaatactctaaccactcggcttttgctgattctggatataaaacaaatgtccaattacgtcctcttgaatttttcttgttttcagtttcttttattacattttcgctcatgatataataacggtgctaatacatttaacaaaatttagtcatagataggcagcatgccagtgctgtctatctttttttgtttaaaatgcaccgtattcctcctttgcatatttttttattagaataccggttgcatctgatttgctaatattatatttttctttgattctatttaatatctcattttcttctgttgtaagtcttaaagtaacagcaacttttttctcttcttttctatctacaaccatcactgtacctcccaacatctgtttttttcactttaacataaaaaacaaccttttaacattaaaaacccaatatttatttatttgtttggacaatggacaatggacacctaggggggaggtcgtagtacccccctatgttttctcccctaaataaccccaaaaatctaagaaaaaaagacctcaaaaaggtctttaattaacatctcaaatttcgcatttattccaatttcctttttgcgtgtgatgcg TO_pHT315: description: pHT315 ~15 copies in Bacillus subtiis. NOTE 897 was mutated G->A to prevent BsaI cut site. Theta replication. - prefix: GTCT - suffix: AAGC + prefix: AAGC + suffix: ATAG tags: [origin] sequence: ccatcctccaaagttggagagtgagttttatgtcgcaaatattaatgtttctggtgaaccttatcaaattttcgttgatttaatagaaacatagcggtaaaattagcagtaacttaatagaacggaaatgaaaaaagccactctcatatgctattggctaccaacctttagcgagaatgacttaatcctgtacagccatacaggacttcgacttataagaggcgccaacttcaaataagttatttgccttgttttcgcgaacaaggcttattagatacacctattgtaccgttactctacgaatatttcagctagtaattactagcattgtcatatacataataaaacggatataaaagggcgttttctatacctagaagtcttgtaaatgtacagggcgtttagatatagagaacgccctttttgtgttccgttccagtggaagctaccactttaaaaagatggtctagtgtagccaatgcaggagagtacactcggatatcagttgtcgttgcattcaactgtctgacgtaagcgaggtaaaggacacaagccttgcataaaacaagcctacgggatgtaaatcctaataatgatgataaccaagacgttagcggcaaaaagtgttgggggttcaaaataagacatgattgtgcgactggagttaaacagttactcgtaagcggcgatcatgacactgattcacggctattcttgtacaagctagctttattacaaggatatgcgggttatatagcgaatcacccgaaagggaacggtgttgggcgtgagaaacgcaccgtacggcgcaatacaatgccaataagctatatacggacggtatagtagttttgtaagctataaccgtttgtcgtcaatgcaaccaatctcaattcAagacctcggcatctaagccagtacgaatgagtgggcgttttaacctcgtaaattttcaacaggggttactatgcccaaaactacattcagatttcctaacaaactcgccagtatgaaaaccttaagaccttaaagtcaagggatttgaaggattttaacctcgattagcaaaaaatgtagagtactgaagcaactaccattaactaagatagtgggggattgaggaagaatccagagctgtttaaatcaagtgaaagacaagatgaaattaaaagaatagtgaaagataggggagtggttctctatgagaaaggaaatggctagagaacaaaggcagcggtttattgatctattgttagactttatggtaaagaatcctcatttatttgttaatggtacagaggatgaaagtaataatgttgttacaaaatgtaatagtgatattaaagaggttgcggagtcatatttaactcttttatagtgagagggttaaaactaattaatatgtattaaggcccaatgttggaattattgtatttcactaggcaacctacttactaaaagtaagattatccattagtggatgttataatattgggttttttaacacaataatcatcgcctttcggtgtcgtttgatagaaaagtaaccattagcgatgaaaaagtcaatataaaaagccatccgtaaaaaacggatggcttaccgtacataggatcgttggtagggcggcgtatcctacatctctggtaacttacctagccaatcaaatgcttgagaacggcggttagataagcgcgtggggaacctttcccacctcaaagatcctatatcattattatgttactttctacaggtagtataccatgttcttatattttagtaaactccccgttagcttaacaggtctttgtaagcaattaaacgtccactattcaatcgtctttggattttcgcaggaccgttttttagatcgaacatagttgataagaacaaataaccgcttgggtccaactttatagcaattagtatatggtcatttaaaatctttaccaattcaacgctattaggttctttaggattttgcccgacatagtcggggtgttcaacgatatcttttatgtgcgatgaatatttttcataaataccaggatgttgtttctttacgtgctttataaatccgggaaacatttttacatcgttagaagtgcaagtcaagttatatgtatctataatgatttgtggaagttttgccacaacagttggtttatttacaatcttttttttattagccgtcaaatttctccctcatctcgtctctttatatctttattttatcataaaggagtatttgaaccgtcgcgcgggacaggtttatgatagggatattttattgaataattgatggtataagggactttcatgcttggaaagtggggattatgaattagatgcttgtccacaatatgttccaatgtaattaaaatttatgttcccaccttgaccaaacatcacgtccatacttaaatcgtccctcctttaataggtaaaatattaatttaccttaataaaaaaataatggataatagtattcgtctgaatttatataatcagggggaactattgatgctggggatactatttacagcggcgccatctactgatgtcgtaaaggatttgcaagataaagttatatcattgcaggatcatgaggtagcgtttttgaacaccacgatatctaatatgttgatcccc pAMbeta1: description: - prefix: GTCT - suffix: AAGC + prefix: AAGC + suffix: ATAG tags: [origin] sequence: gttaactaactcaacgctagtagtggatttaatcccaaatgagccaacagaaccagaaccagaaacagaatcagaacaagtaacattggatttagaaatggaagaagaaaaaagcaatgacttcgtgtgaataatgcacgaaatcgttgcttattttttttaaaagcggtatactagatataacgaaacaacgaactgaatagaaacgaaaaaagagccatgacacatttataaaatgtttgacgacattttataaatgcatagcccgataagattgccaaaccaacgcttatcagttagtcagatgaactcttccctcgtaagaagttatttaattaactttgtttAaagacggtatataaccgtactatcattatatagggaaatcagagagttttcaagtatctaagctactgaatttaagaattgttaagcaatcaatcggaaatcgtttgattgctttttttgtattcatttatagaaggtggagtttgtatgaatcatgatgaatgtaaaacttatataaaaaatagtttattggagataagaaaattagcaaatatctatacactagaaacgtttaagaaagagttagaaaagagaaatatctacttagaaacaaaatcagataagtatttttcttcggagggggaagattatatatataagttaatagaaaataacaaaataatttattcgattagtggaaaaaaattgacttataaaggaaaaaaatctttttcaaaacatgcaatattgaaacagttgaatgaaaaagcaaaccaagttaattaaacaacctattttataggatttataggaaaggagaacagctgaatgaatatcccttttgttgtagaaactgtgcttcatgacggcttgttaaagtacaaatttaaaaatagtaaaattcgctcaatcactaccaagccaggtaaaagcaaaggggctatttttgcgtatcgctcaaaatcaagcatgattggcggtcgtggtgttgttctgacttccgaggaagcgattcaagaaaatcaagatacatttacacattggacacccaacgtttatcgttatggaacgtatgcagacgaaaaccgttcatacacgaaaggacattctgaaaacaatttaagacaaatcaataccttctttattgattttgatattcacacggcaaaagaaactatttcagcaagcgatattttaacaaccgctattgatttaggttttatgcctactatgattatcaaatctgataaaggttatcaagcatattttgttttagaaacgccagtctatgtgacttcaaaatcagaatttaaatctgtcaaagcagccaaaataatttcgcaaaatatccgagaatattttggaaagtctttgccagttgatctaacgtgtaatcattttggtattgctcgcataccaagaacggacaatgtagaattttttgatcctaattaccgttattctttcaaagaatggcaagattggtctttcaaacaaacagataataagggctttactcgttcaagtctaacggttttaagcggtacagaaggcaaaaaacaagtagatgaaccctggtttaatctcttattgcacgaaacgaaattttcaggagaaaagggtttaatagggcgtaataacgtcatgtttaccctctctttagcctactttagttcaggctattcaatcgaaacgtgcgaatataatatgtttgagtttaataatcgattagatcaacccttagaagaaaaagaagtaatcaaaattgttagaagtgcctattcagaaaactatcaaggggctaatagggaatacattaccattctttgcaaagcttgggtatcaagtgatttaaccagtaaagatttatttgtccgtcaagggtggtttaaattcaagaaaaaaagaagcgaacgtcaacgtgttcatttgtcagaatggaaagaagatttaatggcttatattagcgaaaaatcagatgtatacaagccttatttagtgacgaccaaaaaagagattagagaagtgctaggcattcctgaacggacattagataaattgctgaaggtactgaaggcgaatcaggaaattttctttaagattaaaccaggaagaaatggtggcattcaacttgctagtgttaaatcattgttgctatcgatcattaaagtaaaaaaagaagaaaaagaaagctatataaaggcgctgacaaattcttttgacttagagcatacattcattcaagagactttaaacaagctagcagaacgccctaaaacggacacacaactcgatttgtttagctatgatacaggctgaaaataaaacccgcactatgccattacatttatatctatgatacgtgtttgttttttctttgctgtttagcgaatgattagcagaaatatacagagtaagattttaattaattattagggggagaaggagagagtagcccgaaaacttttagttggcttggactgaacgaagtgagggaaaggctactaaaacgtcgaggggcagtgagagcgaagcgaacacttgattttttaattttctatcttttataggtcattagagtatacttatttgtcctataaactatttagcagcataatagatttattgaataggtcatttaagttgagcatattagaggaggaaaatcttggagaaatatttgaagaacccgattacatggattggattagttcttgtggttacgtggtttttaactaaaagtagtgaatttttgatttttggtgtgtgtgtcttgttgttagtatttgctagtcaaagtgattaaatagaattc pBM300: description: - prefix: GTCT - suffix: AAGC + prefix: AAGC + suffix: ATAG tags: [origin] sequence: caagtcctacatcaagtcctaactcgtcaaatagtacatcttcatatagagactcaaaatggttagaaaactgtagggcaaattgttcgagatgtttttggatattatagtagttatcactaaattcctgttttaaaaagatgtcggtaaagcaagtataaccgccgttacctagatttgatacaatacttccctttctacctattcttggatatattgaggtaattacccaatttccatttccatctttttgcacatgtagtctaaaatcatatacatttccagattttattttgcatgttataaacttctgaactaagtaagtaccctcgcttatctgactctcgataaactctattagttggttttcactaagttttaacttttgttcgtttttactaatctcataataattatccgttttctctacaaagaagatacctgcccctttatgtccagataaaggttttacgattatttttttatagcgttcaaccatatctaacaagaccttataactagttagttttgaagtaggaataagatactgggtaaatttcttagcctgattcaccttattataaacggttaatttatcaccaacggagtgactagtgaatgggataatcttgcttaattcatgagttatttttgccattttatcgtttacaggggcgctagcattataaataacatcagggtaagaatattttctctcaacccatttgcctttttcgtaggccattcctaaaataactttttttttcaaatccaccttccctggagtgaaataaaaaaagtctattccttcacctttggcaactatagcaaaagcataagctttttttgcatttattgggtctttacgatgatgaagcatgccgattaaagtcataaaaaccatcctttcccttactcttaattgtttaggtatattcattaatttcatatattatttgtttttgatgaaccctggttgtcctttttttatatcatagattaagagaaggaaatatatgtattttatatgatgaaatcttaggataggatgatagatgtttatggatactattgtacttattgaaacaaataaatctggatcgagtagagaagcaattaaagcagcaaacaaacttaatttctttacggttttgttaactcgcagaaaaaaatttcttgaggaacgaaatgaattcccagatgtacatcaaatgattttcactgatacaaacgattataataatttagttacaataataaaaaagttaaaaaaggaagggaaaaa pBS72: description: - prefix: GTCT - suffix: AAGC + prefix: AAGC + suffix: ATAG tags: [origin] sequence: gaattcatattacttagaggatactattgagagagctattgataagatggttgagacattacctgagagccaaaaaactttttatgaatatgaattaaaaaaaagaaccaacaaaggctgagacagactccaaacgagtctgtttttttaaaaaaaatattaggagcattgaatatatattagagaattaagaaagacatgggaataaaaatattttaaatccagtaaaaatatgataagattatttcagaatatgaagaactctgtttgtttttgatgaaaaaacaaacaaaaaaaatccacctaacggaatctcaatttaactaacagcggccaaactgagaagttaaatttgagaaggggaaaaggcggatttatacttgtatttaactatctccattttaacattttattaaaccccatacaagtgaaaatcctcttttacactgttcctttaggtgatcgcggagggacattatgagtgaagtaaacctaaaaggaaatacagatgaattagtgtattatcgacagcaaaccactggaaataaaatcgccaggaagagaatcaaaaaagggaaagaagaagtttattatgttgctgaaacggaagagaagatatggacagaagaacaaataaaaaacttttctttagacaaatttggtacgcatataccttacatagaaggtcattatacaatcttaaataattacttctttgatttttggggctattttttaggtgctgaaggaattgcgctctatgctcacctaactcgttatgcatacggcagcaaagacttttgctttcctagtctacaaacaatcgctaaaaaaatggacaagactcctgttacagttagaggctacttgaaactgcttgaaaggtacggttttatttggaaggtaaacgtccgtaataaaaccaaggataacacagaggaatccccgatttttaagattagacgtaaggttcctttgctttcagaagaacttttaaatggaaaccctaatattgaaattccagatgacgaggaagcacatgtaaagaaggctttaaaaaaggaaaaagagggccttccaaaggttttgaaaaaagagcacgatgaatttgttaaaaaaatgatggatgagtcagaaacaattaatattccagaggccttacaatatgacacaatgtatgaagatatactcagtaaaggagaaattcgaaaagaaatcaaaaaacaaatacctaatcctacaacatcttttgagagtatatcaatgacaactgaagaggaaaaagtcgacagtactttaaaaagcgaaatgcaaaatcgtgtctctaagccttcttttgatacctggtttaaaaacactaagatcaaaattgaaaataaaaattgtttattacttgtaccgagtgaatttgcatttgaatggattaagaaaagatatttagaaacaattaaaacagtccttgaagaagctggatatgttttcgaaaaaatcgaactaagaaaagtgcaataaactgctgaagtatttcagcagttttttttatttagaaatagtgaaaaaaatataatcagggaggtatcaatatttaatgagtactgatttaaatttatttagactggaattaataattaacacgtagactaattaaaatttaatgagggataaagaggatacaaaaatattaatttcaatccctattaaattttaacaagggggggattaaaatttaattagaggtttatccacaagaaaagaccctaataaaatttttactagggttataacactgattaatttcttaatgggggagggattaaaatttaatgacaaagaaaacaatcttttaagaaaagcttttaaaagataataataaaaagagctttgcgattaagcaaaactctttactttttcattgacattatcaaattcatcgat pCG1: description: - prefix: GTCT - suffix: AAGC + prefix: AAGC + suffix: ATAG tags: [origin] sequence: attgaaggaggtttgtggcaagattttgatttagatcgttagataaattaggggaacctttattcttttctcctcctagaaaaggacttgcatatgcaagtcctcattcttaatacatgttcaaattacaaaaaatcatagatatctgtctgatgtgatgctttctctgcttctatgattacctctgcagtggttggcatccgtctatccctaattaccatctcatacccgataacttttcgtcctttttttacagttgttaaatctatatcggcgttgaatttcatttctaattcttcgcatgccaccatgagagcattcctcttaaataaacctgcaggccatgacttttcttcacccaagaaccaatcttgccattcctctaaacttcccttaatggttgtatattctcgatttcctaagcgatatgcttcccaaagtttcatcaagatcaaagaatactttgatttcacatttcctaactcatccaaatggaaagagtagtagtttcttttcaactcaaaaactaaaggtccagcatcttcagagaatttaaactctacaaggccatcctcaataaatcttattctgctgaaaagttgagtcatgattattgaacgtctaccatcaggcaatgtttcaggaatatatagagctgtattctcattaagtctcgaaaaagcttctgcaatccgtctataattctgaccgtttttagttaagccaaagtgcttcataatgtcaagactatgaaccgtaaagacatcaccaactctactatctttcgttacagatgaaaaacagtagtctaaaaccttgtgttcaaaagcttttaaattaccaaatgctttggccaagtcattcgcctggacaactaaataattttgacgagaagctagttcttcataaatcttacttgattttctatttgataccatcttgtttaccttttactatgttttctattatgctatattctatcaatttttaattttatagtcaataatggggtacaaaatagtcatcaatggggtacaaaatagtcatcaatggggtacaaaatagtcatcaatggggtacaaaatagtcaataatggggtacaaaatagtcaattatttatctgtaaacccttgatattactgagtttctaaggtcgacaaagtaatataaagtaatataaagtaggttagaaagtaaattagaaagtatcttcttaagaaaaaaatctatattttatttttaaaaaataaaaaaatctttcattcagttcactattttctatctaaccactttaaaatataatggctcaaaatctctattctaatatca pCM66: description: - prefix: GTCT - suffix: AAGC + prefix: AAGC + suffix: ATAG tags: [origin] sequence: cgacttccggcaagctatacgcgccctagaattgtcaattttaatcctctgtttatcggcagttcgtagagcgcgccgtgcgtcccgagcgatactgagcgaagcaagtgcgtcgagcagtgcccgcttgttcctgaaatgccagtaaagcgctggctgctgaacccccagccggaactgaccccacaaggccctagcgtttgcaatgcaccaggtcatcattgacccaggcgtgttccaccaggccgctgcctcgcaactcttcgcaggcttcgccgacctgctcgcgccacttcttcacgcgggtggaatccgatccgcacatgaggcggaaggtttccagcttgagcgggtacggctcccggtgcgagctgaaatagtcgaacatccgtcgggccgtcggcgacagcttgcggtacttctcccatatgaatttcgtgtagtggtcgccagcaaacagcacgacgatttcctcgtcgatcaggacctggcaacgggacgttttcttgccacggtccaggacgcggaagcggtgcagcagcgacaccgattccaggtgcccaacgcggtcggacgtgaagcccattgccgtcgcctgtaggcgcgacaggcattcctcggccttcgtgtaataccggccattgatcgaccagcccaggtcctggcaaagctcgtagaacgtgaaggtgatcggctcgccgataggggtgcgcttcgcgtactccaacacttgctgccacaccagttcgtcatcgtcggcccgcagctcgacgccggtgtaggtgatcttcacgtccttgttgacgtggaaaatgaccttgttttgcagcgcctcgcgcgggattttcttgttgcgcgtggtgaacagggcagagcgggccgtgtcgtttggcattgctcgcatcgtgtccggccacggcgcaatatcgaacaaggaaagctgcatttccttgatctgctgcttcgtgtgtttcagcaacgcggcctgcttggcctcgctgacctgttttgccaggtcctcgccggcggtttttcgcttcttggtcgtcatagttcctcgcgtgtcgatggtcatcgacttcgccaaacctgccgcctcctgttcgagtcgacgcgaacgctccacggcggccgatggcgcgggcagggcagggggagccagttgcacgctgtcgcgctcgatcttggccgtagcttgctggaccatcgagccgacggactggaaggtttcgcggggcgcacgcatgacggtgcggcttgctatggtttcggcatcctcggcggaaaaccccgcgtcgatcagttcttgcctgtatgccttccggtcaaacgtccgattcattcaccctccttgcgggattgccccgactcacgccggggcaatgtgcccttattcctgatttgacccgcctggtgccttggtgtccagataatccaccttatcggcaatgaagtcggtcccgtagaccgtctggccgtccttctcgtacttggtattccgaatcttgccctgcacgaataccagctccgcgaagtcgctcttcttgatggagcgcatggggacgtgcttggcaatcacgcgcaccccccggccgttttagcggctaaaaaagtcatggctctgccctcgggcggaccacgcccatcatgaccttgccaagctcgtcctgcttctcttcgatcttcgccagcagggcgaggatcgtggcatcaccgaaccgcgccgtgcgcgggtcgtcggtgagccagagtttcagcaggccgcccaggcggcccaggtcgccattgatgcgggccagctcgcggacgtgctcatagtccacgacgcccgtgattttgtagccctggccgacggccagcaggtaggcctacaggctcatgccggccgccgccgccttttcctcaatcgctcttcgttcgtctggaaggcagtacaccttgataggtgggctgcccttcctggttggcttggtttcatcagccatccgcttgccctcatctgttacgccggcggtagccggccagcctcgcagagcaggattcccgttgagcaccgccaggtgcgaataagggacagtgaagaaggaacacccgctcgcgggtgggcctacttcacctatcctgcccggctgacgccgttggatacaccaaggaaagtctacacgaaccctttggcaaaatcctgtatatcgtgcgaaaaaggatggatataccgaaaaaatcgctataatgaccccgaagcagggttatgcagcggaaaagatccgtcgaccctttccgacgctcaccgggctggttgccctcgccgctgggctggcggccgtctatggccctgcaaacgcgccagaaacgccgtcgaagccgtgtgcgagacaccgcggccgccggcgttgtggatacctcgcggaaaacttggccctcactgacagatgaggggcggacgttgacacttgaggggccgactcacccggcgcggcgttgacagatgaggggcaggctcgatttcggccggcgacgtggagctggccagcctcgcaaatcggcgaaaacgcctgattttacgcgagtttcccacagatgatgtggacaagcctggggataagtgccctgcggtattgacacttgaggggcgcgactactgacagatgaggggcgcgatccttgacacttgaggggcagagtgctgacagatgaggggcgcacctattgacatttgaggggctgtccacaggcagaaaatccagcatttgcaagggtttccgcccgtttttcggccaccgctaacctgtcttttaacctgcttttaaaccaatatttataaaccttgtttttaaccagggctgcgccctgtgcgcgtgaccgcgcacgccgaaggggggtgcccccccttctcgaaccctcccggcccgctaacgcgggcctcccatccccccaggggctgcgcccctcggccgcgaacggcctcaccccaaaaatggcagccaagctgacccgctagggacgtgaagtcgattccttcgatggttagcaatcaaagactcagagtgctgtctggagcgtgaatctaacggtacgtatctcgattgctcggtcgctattcgcactctgcgaaagttcgtaccgctcattcactaggttgcgaat pGRB1: description: - prefix: GTCT - suffix: AAGC + prefix: AAGC + suffix: ATAG tags: [origin] sequence: GATCTactcatcgacatcccaatctgctgtagctcacctacatcgctctcatcgcgtccgctctacgacccacttcttcaacccactccgtgggcaaggccagaccgccgtctgagcgtcttaaccacccggtggcggcgttcgcaacctcctgtacgggtcgctccgtggctccgacgggtccacctcgatgatgtccccgagatccccctccggcgcgatgcccaccatctcccactcctcgatgctgtcgccctcgtcgtcctcctccggttgcatcagctcctgtgcgccgttgctcggacggaaccactgccgaccactcgcccacatcgtcgcgtagaatgcacggacgttctcaggcatctcagacggctctgagccgtactcgccggccatgtacgccgccaggtacgccccgaggttctcgacgccaccgctgcgcgacgggtgcgacgagcgccgaactctcaccgcgtcctcgtcgccgttctcgtcgaacacctgatgggcgtcctcgcccgccgtcgggcagtttcgcagatgcgcgtccagcaccggctcgaactgctcggcgaccaccggccccctcacaaacacgccaagatggatgtgtacgtacccgctctcgtgcggttcaaggatggcgaggtactcccactctctaccctccaaaactcgcgccagcgcccgcctaacagcctcccacgactccagcaggtcctcgaagtgttccagcggaggccgcagccgtccctcgtcgtccgtactcgacgccgtgagcgtcaccatccccgtgtgcagaaggctcccccaccgctcccgaaggccccgttcgaggctctgcgccttcgcgtacagcatctcgcggtactccggcgtgaagcggttcggcatgtcaaagcccaccgtctcgcccagctcgttctcgacgacgatctgcgttccacgctggtcgtcgtaccagctctggaagcggtcgatggcctccccccacgacaccgccttgacggcccggctcgtgtcgaaggtcagttcatcccgcagtttcatcccgtctcgtttcgccatcggcacgtgtgaggcgctgaggctgtcgttctccgggcctgcacggttattcaaaggttgccgagaaccgcccgagttggcaggccggtcgggcggttcacccggctgattaggtgtgctcaccggaacccctccgttccgggaacgaggacggccgcgttgcggcctgtgttccgggctgcgggccgctgcgcggcccgttccgggctttcacccatcccttcactgcggggatgctggacagcacgggagatcccccgcctgccggctgttgcggcccctgagagggacctggggcacatataccagccgtctcgccgtcgggaaagacctaagcggacggacgaagaagggccgagtgaaccagccaggcaaggtggttcgtttgccggttgcaggccgcgggtacggccccttccggttttgtcccggcccttctccggggcacctttcacgttgcgtagctgttactccaccgtcattcagctatccagttattagttgcggcgtggagccagtattggtccttaat pIP404: description: - prefix: GTCT - suffix: AAGC + prefix: AAGC + suffix: ATAG tags: [origin] sequence: gaattcttttaaggggcttatagattatctattttaaataaaaaggattcttttaatttattattaaaagaatcctttttattaattatttttaactgaattaatgtatatatatatatgataaaatccaaatgtaaaaatgcaagaaaaaacatacattaaaaagcttataatataatattgtggaatattgaaattaattttactaaaaactattccaatcaaggcagcaataataaaaatagttttatttattggtttagagtacgtgtttttatttttaagtttctttttaatagctgttagaaagaaacttaaaaataaaatatatataataataaatataataaaaatatatataaatttagaaaaagataatatttgataaaacatatttaaagatataatcaaatcctgtaaagatgaaaatgaaaataaaattcctaaaaataagtttaataaagccggaaatttcactagtttttttaaaaacaataaatataataaagatattattataaaacttaaatttaataataacatcataaggaaaatggattggtttacttcaacggctttattatatgaaaaaaatacaattaaatttaaacataatattacaaaaagtattgttaaaattgaaaaaattaaatctttagtatctttggaaataatcataatttataagctcctttgattttttatataaattatacattaattatttaatatataaaaactaaaattagtaaatcatctaaatattacaaatgaagaaatattatttgtaaataattatagtaatatattatggaatgaaatagtaaaaaattaaaaagaatcatttaatttagtttaaaatattttaataaaaataataaattaaaacataaaaaatagattaagtttcaattggggacttactctattttttattaagagtttaagaatagttaataattattgaaaatatgatataattttcttagttttacagaaggggaggtgaataagtgagaaacgcattaaagctgaataaaaagaactatatagataatattcatagtagaagtaaagggtggataaccaggagcgttatagataaaaaaggatatagccaatggcactataaatacgctgaattaaaagatttagatatgagtgatgaaaatatctatataactctaaataccttttataagccgtgtaggcgattagaaaatataaaagagttaaatacactgtttatagacttggattattataaaactggcaaaactaaagaccaggtattaatggacttagaaaagaattattttaatcaaagtattcctataccaaactatgtaatagatagtggaagaggaatgtatttaatatggataataaatgcagtacctagtaaagcattaccattatggaaagcggttcaagaatatttatataatcaattaaaatactttggagcagatagacaagccttagatgcaaccagaatattaagagttccaggaagtataaactctaaatcaaaaacagtagtcaatatattagatgagtacgaatatatttatgacttaagagaaatacaaaatggatttttacctgaattaaaaccatatgaaaggaaaaagggtagaccaagcaaaataaattatatttatagagaaagaagtttatattatggaagaatacaagacataataaaactttgtgaactaagagaatatgatttaaaaggacacagagagcttatattatttttatatagatattatctttgtagctttacagaggacattgagaaggcattaaatgatgttttagaacttaatagtatgtttagacaacatttaagtgaaagagaagttataagagcaactagaagtgctgaaagatgttatttagataaaaataagcaatataagtataagaatgaaactcttatagagttattagaaattacagaagaagaacaaaaatatatgacaataataatttctaaaaaggaatataagagaagagaaaatattagaggtaaaaaaaattatcaagagcaattaaaagctaaaggaaaagcaacaaaaaaagaggaattaaatgtattaagaaaaaaaataaaagcccttaaagaaaaaggctttaaaaataaagaaattactctaatgttagaagtaccaataaaaacattagaacgtcatattacgtatatgaaaaaaaatgggcttttataaaggctcattttttatattcttttcttcaaagattatataatataaaaaaatttttttcaaactttaaataaaaaatatttttatatttttttatttttttatttttatatttttttatttttttatttttatatttttttatttttatatttttttatttttatatttttttatttttatatttttttatttttttatttttttatttttttatttttttatttttttatttttttatttttttattttttttatttttttatttttttatttttttatttttttatttttaccctcatttttttacgcttgtattatagggtactttgtacctgttcttttttttggggaggttgtaaagataattttttactttagttagttccgaaggaacttttattatagtgcccctttatttttttgcgtggcattgaatgtaaaaaattatcactatactagggcgtaaagtaatattacatgtgtctcaaagtgggattaaagcgggattttatagggcgtgtttgtggcttagagtgggattattggaaatttttttgatatc pLS20: description: - prefix: GTCT - suffix: AAGC + prefix: AAGC + suffix: ATAG tags: [origin] sequence: ATCATTACTAcaggcccggggctttacgtcaaaaaaaaaatcgtgtaagataaagtcaagttaagtcaatgtgaaagttttggcattgaattctggttctagttattctaaataaacaacaaaaaaccctgttgtttgcagacaacaaggttaatagatttgatgatacatattttgtttgcgagctatttgcaaaattgaatacaaataaagaatgacaaataaaaagtgtccgatgttgcagcatcgaacacttggtttgtaagccctcacttacaatgtttaagttatAtcttcattttaccgaaaacaagtgtatacgtcaacacttttcggctatttttggacgacagaaaaccactgtttggagggtatcaaaccgtggttttctgtcgttttttgtttgtcacccagctgccgttaaggggtgtaaaataaactgacgtcaggagttgccccgtccaccgcaaacccggggatataccacaataagcgttccttgctgcgcttgtggtgaatggcgaggacggccattaacggtccggagaatgtggataaaccactaggttgcttggcgcactacggtgcgggcaactatggccaagcttctatgcagcggcacacaattgagtgtgaccgtgtgagagaacacgagacaaaccccatagaagttgcatacaggacaagcattgtatgtaaaaaggtcatggcaggcgaaagcctggcgatagcaaggcagagctggaaacctcagaTgtctctatgacgctagtttcaaatcctgatcgattgcctatgtctgtccgtttttattttttgcgcgacagacataggcaatcgaaattcgccctgtttcctagaacagtcagctattctgctagtttcaagttctcaaagtcaagagaaaaacactaaataaaatgcgaatttccttaaaacccagaggttttaagttcctttttaagggttgttcgtttgatagaacattagtattttaatatatttttcctgtgggatttcactaacgtatactgttattccctttggcctatcttaaaaacttgtatagattgtgtttgagccgtttagtatgatatcagtacatgcttggg pMflST: description: - prefix: GTCT - suffix: AAGC + prefix: AAGC + suffix: ATAG tags: [origin] sequence: CCGTGAACACGAGCGTGTTTAGTTTTTGATGGTTGTCAAGTTCTTTTCATtgtattcaaacccccttttaaagttgataattcttgttaattataatataagtgctggaaaaagtatagttttttattgttatttttaagaaaatttatccattgtgtttttatatgaactttttcaacattttttataaccttttgtggaaaagcggaaaaaagatatataaatactaaactattgctttttagtgttattcattatgtttatttctttttaaaatatttttttatccacacattgtggaaaagttttattattatgtgttttaatttattttatgaaggggaattaatatggaaacaaaagcgctttgagaaaaattaattaataagttaaagaaggaaaaattaatagaccaagacattattgaagaacatattgttacttcagaattaattaaaatttcaaatacagagttcgtaatccttgttagaagtaatcttggtgtaactattttaaatgaatttaaagaagtatttgtgtacgaatttaaataagttttaaacagttatgtgtctgttgattttttaactaaagaaatttttgagaaaaatacaaaaaaagagaataaaaaagaacccataaatactgttttatctgaaaatgcacttacttttgagaattttatagttgggtcaagtaataaacaagctaatttagctgctaaaaatgttgttgcaaaccctggtatgagttttaatcctttatttatttatggagattcaggactgggtaaaacacatttattacaagctattaaaaatcaagcagaattaaatggaaaaaaagttttatatttaacatctgaagaatttacaaaaagaattgttaatgctttaaacaaaggtgatttaagtgaaatagaagaattaaaaactgagataaattctaatgaattctttattttagatgatgttcaatttttaagtaaaaaagataaaacaaacgagtttttctttaacattattaacaattttactgaaaatggtaaacaattagttttttctagtgataagactcctgaattgttaaatggttttgataaaagaatgataactagatttaattcaggtttatcaacacctattaatgctttagatattccaactgccaaattgataattgaagctgaaattaaaaaacaaggtttaaaacaaaaaattaaagaagatgccgttgtttacttagctcaaaactttagcgacgatgttagaaaaattaaaggtttagttaatagattacttttctttggtattcaaaacgatttaggtcacataattgatttggaagatgttattgacttatttaaagatacaccttcagctaatttaggattattaaatgttaaaaaaattaaagaagttgttgctaaaaaatatgatgttactataaaagccatcgatggtaaagcaagaacaactgctataaaaaatgctagacatctttctatgtattttgcaaaaataattttaaaccatacatcaactcaaattggtgcagaatttggtggaagagatcatagtacagttttaagtgctatttcgcgtattgaaaaattaatatataaagaaaaagaattcaaaaaaatagttgagtctttaaaaaacgaaataataggaaaatagatgtaaattcgcatctattttttttgtgtggaaaagaaattaaacttgttgaataaaaagtcatttaaacctttatttaccgttgttttatgagtgttttccacatatcaactgcattataaatattacttatataaagaataatataatatataaaagcatgcgtatttcttttgtttttattttataaatatgagataatatatattgtaaataagaggtgttatatgag pMK3: description: - prefix: GTCT - suffix: AAGC + prefix: AAGC + suffix: ATAG tags: [origin] sequence: aataaaaaaagcacctgaaaaggtgtctttttttgatggttttgaacttgttctttcttatcttgatacatatagaaataacgtcatttttattttagttgctgaaaggtgcgttgaagtgttggtatgtatgtgttttaaagtattgaaaacccttaaaattggttgcacagaaaaaccccatctgttaaagttataagtgactaaacaaataactaaatagatgggggtttcttttaatattatgtgtcctaatagtagcatttattcagatgaaaaatcaagggttttagtggacaagacaaaaagtggaaaagtgcgtccatggagagaaaagaaaatcgctaatgttgattactttgaacttctgcatattcttgaatttaaaaaggctgaaagagtaaaagattgtgctgaaatattagagtataaacaaaatcgtgaaacaggcgaaagaaagttgtatcgagtgtggttttgtaaatccaggctttgtccaatgtgcaactggaggagagcaatgaaacatggcattcagtcacaaaaggttgttgctgaagttattaaacaaaagccaacagttcgttggttgtttctcacattaacagttaaaaatgtttatgatggcgaagaattaaataagagtttgtcagatatggctcaaggatttcgccgaatgatgcaatataaaaaaattaataaaaatcttgttggttttatgcgtgcaacggaagtgacaataaataataaagataattcttataatcagcacatgcatgtattggtatgtgtggaaccaacttattttaagaatacagaaaactacgtgaatcaaaaacaatggattcaattttggaaaaaggcaatgaaattagactatgatccaaatgtaaaagttcaaatgattcgaccgaaaaataaatataaatcggatatacaatcggcaattgacgaaactgcaaaatatcctgtaaaggatacggattttatgaccgatgatgaagaaaagaatttgaaacgtttgtctgatttggaggaaggtttacaccgtaaaaggttaatctcctatggtggtttgttaaaagaaatacataaaaaattaaaccttgatgacacagaagaaggcgatttgattcatacagatgatgacgaaaaagccgatgaagatggattttctattattgcaatgtggaattgggaacggaaaaattattttattaaagagtagttcaacaaacgggccagtttg pMTLB: description: - prefix: GTCT - suffix: AAGC + prefix: AAGC + suffix: ATAG tags: [origin] sequence: Aaattcatattacttagaggatactattgagagagctattgataagatggttgagacattacctgagagccaaaaaactttttatgaatatgaattaaaaaaaagaaccaacaaaggctgagacagactccaaacgagtctgtttttttaaaaaaaatattaggagcattgaatatatattagagaattaagaaagacatgggaataaaaatattttaaatccagtaaaaatatgataagattatttcagaatatgaagaactctgtttgtttttgatgaaaaaacaaacaaaaaaaatccacctaacggaatctcaatttaactaacagcggccaaactgagaagttaaatttgagaaggggaaaaggcggatttatacttgtatttaactatctccattttaacattttattaaaccccatacaagtgaaaatcctcttttacactgttcctttaggtgatcgcggagggacattatgagtgaagtaaacctaaaaggaaatacagatgaattagtgtattatcgacagcaaaccactggaaataaaatcgccaggaagagaatcaaaaaagggaaagaagaagtttattatgttgctgaaacggaagagaagatatggacagaagaacaaataaaaaacttttctttagacaaatttggtacgcatataccttacatagaaggtcattatacaatcttaaataattacttctttgatttttggggctattttttaggtgctgaaggaattgcgctctatgctcacctaactcgttatgcatacggcagcaaagacttttgctttcctagtctacaaacaatcgctaaaaaaatggacaagactcctgttacagttagaggctacttgaaactgcttgaaaggtacggttttatttggaaggtaaacgtccgtaataaaaccaaggataacacagaggaatccccgatttttaagattagacgtaaggttcctttgctttcagaagaacttttaaatggaaaccctaatattgaaattccagatgacgaggaagcacatgtaaagaaggctttaaaaaaggaaaaagagggtctgccaaaggttttgaaaaaagagcacgatgaatttgttaaaaaaatgatggatgagtcagaaacaattaatattccagaggccttacaatatgacacaatgtatgaagatatactcagtaaaggagaaattcgaaaagaaatcaaaaaacaaatacctaatcctacaacatcttttgagagtatatcaatgacaactgaagaggaaaaagtcgacagtactttaaaaagcgaaatgcaaaatcgtgtctctaagccttcttttgatacctggtttaaaaacactaagatcaaaattgaaaataaaaattgtttattacttgtaccgagtgaatttgcatttgaatggattaagaaaagatatttagaaacaattaaaacagtccttgaagaagctggatatgttttcgaaaaaatcgaactaagaaaagtgcaataaactgctgaagtatttcagcagttttttttatttagaaatagtgaaaaaaatataatcagggaggtatcaatatttaatgagtactgatttaaatttatttagactggaattaataattaacacgtagactaattaaaatttaatgagggataaagaggatacaaaaatattaatttcaatccctattaaattttaacaagggggggattaaaatttaattagaggtttatccacaagaaaagaccctaataaaatttttactagggttataacactgattaatttcttaatgggggagggattaaaatttaatgacaaagaaaacaatcttttaagaaaagcttttaaaagataataataaaaagagctttgcgattaagcaaaactctttactttttcattgacattatcaaattcatcg pNG168: description: - prefix: GTCT - suffix: AAGC + prefix: AAGC + suffix: ATAG tags: [origin] sequence: accatagagttgagtagaatatttgacgaatgggaaacacatcatcgaaatttggtggttctacactgtgcgggatcgtcagacaccgactgttacctttaagttgcatccgtgaaaacaaggggccaaggcaggacagagtaatcactggtcggcgttggaagcctgaagaaggccccggacacgaacctctcgtgtccccacaggttgctacctcggccttcttccgctgtgggcttaagcgttaccccacgtagccgctgcgctgccaaatgaaatgtcctcatcacgatgacagcgatgcaaactacacagacacaccaagaccgaggtgaccgcgagtagctgacgctactccagaccaatgactcacaggtacagactcaaacgacaatccgggtccgatgctgccaggcttcgccccggatgtcgttgtgaagcatcgtcgcagcggaccagcagagccggtcctcaacacggatggagcggtattatcagtacgctcctatctacaatgtggacgcgaggggtcaaattggtttcgggaccctcgcatttgccggcttccacgcagtccgcacttctccacggactgcatgcacacgcctaatcaacaacaaggaatccggaagatcgttccaggagggaccctctctactgctggaattaccattaccgaggtcaccccacgggtaacagaatggatacctgaccttcttgaggaactactgccacggagtattcaatcggttcgaaaattcatccggcaagaggacccagaagtcctcacgcacgcacgatacaacaccgtctatcgccgacttcaagaggaaaccctgaggttcgaccatcaagaatggtgctccacaacggatatttggagtgatgcagaggctgaagcggttgaatacgtagaatcacttgtcgagttcgcagtcaaatattctgacgttgacgaagatgatctcgacgaactttcggagtatcaccagcagcggtgcaaatcgcttaaacagactctcactacgatcagtaccggacgtggcccactcaacgctggccttgaagccctcgccaagggacccgtacgactccacgatgagctcgatgacgcaccgcaaccgatcacgcttgtccttgatggcgagttgtggtcaaaactcgacgatagaggaacaggtatccgagcacttgcagctatcgccgtgctcggctccaccttcgacgtccgcctggttatttcaccagcgttagacgccgcgattgagcgacggtatccagactggtatgactcccatctccgtcttactgaaacccgtgaaacctcctctgtagaatcagcgggtggcgacggacaaccatcggcggagcagctcgaggaagcatgggaagcaatacagaacctcccagaggaatcagggagactccgacttctccggaacctccctatagagggctctcgagactatcgtgatctcaagcaggatgatgagattgacgtgcaggcagggactgtcggccgttacatcctcgatctagaagaattagggcttgtggatatcgaccgtcgtggacaatacaacagcgcatccctcaccggcttaggacaagtagcagttgagcagtatgtcaccacggactaccgggtgatccatccgacccaatcgacgctggaaacgcatcttacgccgacccctcagccccaagcaagtacagtgtatcccgcgcgatcggacacgagggagggggatcagcctgggacagcggaggattggatagctgcgacaggcagtcctagtgagggtgctgactacgttcaatggctcgatgggccgtctggtgttctcgacgcttggggaatgcatcagcggtaccttgctggccgtcgagatcgtggtgtcaccctagtcgatgaccgtatcgagcgtttcgaggacggtcgcgtatcgtatctcagttgcttcgacgatgatctcttcgtagccactcagtggggcggcccccttcctaccctgggacgtattgcgggtgccttactcagcgacaaagcactgagtaagatccttaccccttctcgactcggaaaccagttcgaagaaatcaacgatgcggtcgttgaacaactcgatagagaagctggggaaatcatccgtagaggacaccaaatcggctggttcagcgaggacgaagaggactacgatggttggcgtgagcgtattggctccgttcgcagtctgtgtttgcaacaagttggtgaactcacaaacagtgacgacgtcgaagcccggactgagttactacgtgacctgcacggcttggttgcctcagcgacacagctgtactacgcagccggtgtagacgtcacaatcaacgttcgagtccctgacacgggaatgctcatctcggatgaacgccgactcgacgatttcctcggtttcgcccggtacaccattccgaaacagtccgtgtacgggattcattcggggtataggatgctcctcgaggatcgaccggagaaactcaaacgacgacttccctacgaggtggatgatgctgattccacaatgcatctgaccgcatcgtgggttttctccggatcaacgatgatcgatcttcacgatgacatcgaggacgccattgaaatggaaaccaacgaaattcgtgaagcaattgctaatggacaggagtcagcaccggtaatggaaatccccgtccagattgggaactcctactcagcaattcgtaatcacgtcgaggactacgcttcagcgaagaactaccaggtagctcaccaggaggatattcacgaaggaaaacaggatctcgagcgactcgtccgcctgttcctccgtgtcctcgggacagaggatcgaccacatcgagcgtgtcctcacgacgttgccgaggcaatgcttcatgttgctcaatcctcccggaactacgatttcatcaccgtgcgagacatctcgtacggactgtcgaatctccccacgaaacgactcttacccgagctcccaccaacagcaacgaagctcctcaaaaccctgcttgatgcggatgacccgatgggacggtctgaaatcatcgataccgctgacatttcggaaagtagctatgatcgctacatcaacgaactcgccgcttgggatatcattgaaccccgcgaaatcgagggacaccgtcggtgggaagctcacttagagccgtggtggacaccgcagagtgatcgtgacgaaccctatgccgaccctgaccccgacacgggaatactgtacgctgaatttccccgtgatgtcgctagtgcggtgatgtgccacctcatcacccactacgacttacccgaccttgagacagcgtatcttgagggtatccaaccgggggacgacatcaaggccctcttcgatgatcacgaccgactcagacgatggcggccattcctctggggtgcattcgccgactcggacaaacttgaaagaggcccatctggtacagcagcttcggactcgaccgtagttcgactcggtcaatctccaggacccgacacagcacaatcgagcttccaagatgtctcagaaacagcaacccaacgagatcgactcagtcaaccgtcgccagggctggactaatgattgacggcaatcgataactacgcaatgaatcctccacaaccagcatctgaaccgtactcgaaaggtgatgcagtaaccgtatacgtgggc pSK41: description: - prefix: GTCT - suffix: AAGC + prefix: AAGC + suffix: ATAG tags: [origin] sequence: gatttatcggcctcataattttagatatattaattatattagagatttgttatcaaatacatttcattacaagcatataattgacctgtgaggcaacaaaaaatttttaatttttttaaaatagtgttactattgaacatagaaaaagacacctaacgcttggcggttgttagatgtctaaagagtttaaataatatgtgttatttacttttacaatttgtttgtatctttattatatatacaacctcgatacattacaagataaaagcaacaagtctttaagttatatataacggataacattatataaccttattcatcacttgtaaataacgatattacttaaactccttaaatatacatttaaaggagtttttatcatgtctaaacaattttttacagtagaagaaaattataaagaacgcttttaccaattacctaaagtattttttactaatcctaattataaagatttatcgaatgacgctaaaattgcttacgcaatattgagagatcgtttacaattatctattaaaaataactggatagatacagaaggtaacatatatttcatttatacagttgctgatttagaagtgatattaaattgtgggaacaaaaaaatcactaaaattaaaaaagaattagaaaatgtagatttactaatccaaaaaaggcagggtttaaacaaaccaaatctactttatttattaaaaccagccataactaaaaatgatatttatgaaatagataaggctgaaaatgaagttgaggccttacaagacaaggaagtgtcaaaaggacacgtccagaaatgtcaaaaggacacgtctagaaatgtcaaaaggacacgtctagaaatgtcaaaaggacacacaaatgatactgattttattgatactgattttattgatactgaaagtaatgatatgaataatatgaatgatacaaatcaacattcaaatcattcaaatcacttttcaaatattcatgataaagaatctcttaaatatatagaacttcaagaactccccgaattaattaaaagttatattaataatttcagctatgaagaagttaaaagtattaaatctgttattttaaaagctaaaaaatcatttaataacaaatacgatactttctatatgttagaagatatagatgaagaattactattggttttaaaacgatttaaaggttatcttgttaaaaaacaagaaaaagtagctaatatggaaggttatttaatgagaagtatcattgctgaacttgaagaaatgcactcaactattatgagaagaaaaaatatggaaaacaatccattatctctatttaattagtttttgaagttaaattttcaaaaaaaagagattatacacctaaaaaggtgtataatctttac diff --git a/parts/parts/meganucleases.yaml b/parts/parts/meganucleases.yaml new file mode 100644 index 0000000..5a9a690 --- /dev/null +++ b/parts/parts/meganucleases.yaml @@ -0,0 +1,40 @@ +# notes: added GSHHHHHHHH to each end of protein. +fuGFP(Ec)-gs8xhis: + tags: [protein] + sequence: MVSSGEDIFSGLVPILIELEGDVNGHRFSVRGEGYGDASNGKLEIKFICTTGRLPVPWPTLVTTLSYGVQCFAKYPEHMRQNDFFKSAMPDGYVQERTISFKEDGTYKTRAEVKFEGEALVNRIDLKGLEFKEDGNILGHKLEYSFNSHYVYITADKNRNGLEAQFRIRHNVDDGSVQLADHYQQNTPIGEGPVLLPEQHYLTTNSVLSKDPQERRDHMVLVEFVTAAGLSLGMDELYKSGSHHHHHHHH +I-SceI(Ec)-gs8xhis: + tags: [protein] + sequence: MKNIKKNQVMNLGPNSKLLKEYKSQLIELNIEQFEAGIGLILGDAYIRSRDEGKTYCMQFEWKNKAYMDHVCLLYDQWVLSPPHKKERVNHLGNLVITWGAQTFKHQAFNKLANLFIVNNKKTIPNNLVENYLTPMSLAYWFMDDGGKWDYNKNSTNKSIVLNTQSFTFEEVEYLVKGLRNKFQLNCYVKINKNKPIIYIDSMSYLIFYNLIKPYLIPQMMYKLPNTISSETFLKGSHHHHHHHH +I-SceI(site): + sequence: TAGGGATAACAGGGTAAT +I-CreI(Ec)-gs8xhis: + tags: [protein] + sequence: MNTKYNKEFLLYLAGFVDGDGSIIAQIKPNQSYKFKHQLSLTFQVTQKTQRRWFLDKLVDEIGVGYVRDRGSVSDYILSEIKPLHNFLTQLQPFLKLKQKQANLVLKIIEQLPSAKESPDKFLEVCTWVDQIAALNDSKTRKTTSETVRAVLDSLSEKKKSSPGSHHHHHHHH +I-CreI(site): + sequence: GAAACTGTCTCACGACGTTTTC +PI-SceI(Ec)-gs8xhis: + tags: [protein] + # notes: From https://www.addgene.org/195610/ + sequence: MCFAKGTNVLMADGSIECIENIEVGNKVMGKDGRPREVIKLPRGRETMYSVVQKSQHRAHKSDSSREVPELLKFTCNATHELVVRTPRSVRRLSRTIKGVEYFEVITFEMGQKKAPDGRIVELVKEVSKSYPISEGPERANELVESYRKASNKAYFEWTIEARDLSLLGSHVRKATYQTYAPILYENDHFFDYMQKSKFHLTIEGPKVLAYLLGLWIGDGLSDRATFSVDSRDTSLMERVTEYAEKLNLCAEYKDRKEPQVAKTVNLYSKVVRGNGIRNNLNTENPLWDAIVGLGFLKDGVKNIPSFLSTDNIGTRETFLAGLIDSDGYVTDEHGIKATIKTIHTSVRDGLVSLARSLGLVVSVNAEPAKVDMNGTKHKISYAIYMSGGDVLLNVLSKCAGSKKFRPAPAAAFARECRGFYFELQELKEDDYYGITLSDDSDHQFLLANQVVVHAKWVADPNSSSVDKLAAALEHHHHHHHH +PI-SceI(site): + sequence: ATCTATGTCGGGTGCGGAGAAAGAGGTAATGAAATGG +I-TevI(Ec)-gs8xhis: + tags: [protein] + # notes: from https://www.researchgate.net/profile/Marlene-Belfort/publication/13964870/figure/fig2/AS:601631284527125@1520451484869/GIY-YIG-endonuclease-I-TevI-The-bipartite-homing-site-is-shown-above-with-the-two.png + sequence: MKSGIYQIKNTLNNKVYVGSAKDFEKRWKRHFKDLEKGCHSSIKLQRSFNKHGNVFECSILEEIPYEKDLIIERENFWIKELNSKINGYNIADATFGDTCSTHPLKEEIIKKRSETVKAKMLKLGPDGRKALYSKPGSKNGRWNPETHKFCKCGVRIQTSAYTCSKCRNRSGENNSFFNHKHSDITKSKISEKMKGKKPSNIKKISCDGVIFDCAADAARHFKISSGLVTYRVKSDKWNWFYINAGSHHHHHHHH +I-TevI(site): + sequence: TATCAACGCTCAGTAGATGTTTTCTTGGGTCTACCGTTTAATATTGCGTCATATGC +I-AniI(Ec)-gs8xhis: + # from: https://pmc.ncbi.nlm.nih.gov/articles/PMC2040299/ + # uses lib4, a better binding site + tags: [protein] + sequence: MRILKSHPLLKIVNSYIIDSPQPANLSYLWNFGSLLALCLGIQIVTGVTLAMHYTPSVSEAFNSVEHIMRDVNNGWLVRYLHSNTASAFFFLVYLHIGRGLYYGSYKTPRTLTWAIGTVILIVMMATAFLGYVLPYGQMSLWGATVITNLMSAIPWIGQDIVEFIWGGLYTDEPQCGDVLLKILLNAGKSPILGFAYDLFFIIVLLIGVKIAMTRGKSAGVRSLHTSEASQRLHAGDLTYAYLVGLFEGDGYFSITKKGKYLTYELGIELSIKDVQLIYKIKKILGIGIVSFRKINEIEMVALRIRDKNHLKSFILPIFEKYPMFSNKQYDYLRFRNALLSGIISLEDLPDYTRSDEPLNSIESIINTSYFSAWLVGFIEAEGCFSVYKLNKDDDYLIASFDIAQRDGDILISAIRKYLSFTTKVYLDKTNCSKLKVTSVRSVENIIKFLQNAPVKLLGNKKLQYLLWLKQLRKISRYSEKIKIPSNYGSHHHHHHHH +I-AniI(site): + sequence: TGAGGAGGTTACTCTGTTA +I-DmoI(Ec)-gs8xhis: + # https://doi.org/10.1007/s10822-017-0087-5 + # https://www.jbc.org/article/S0021-9258(20)69746-0/fulltext + tags: [protein] + sequence: MHNNENVSGISAYLLGLIIGDGGLYKLKYKGNRSEYRVVITQKSENLIKQHFAPLMQFLIDELNVKSKIQIVKGDTRYELRVSSKKLYYYFANMQERIRLFNMREQIAFIKGLYVAEGDKTLKRLRIWNKNKALLEIVSRWLNNLGVRNTIHLDDHRHGVYVLNISLRDRIKFVHTILSSHLNPLPPERAGGYTGSHHHHHHHH # I52F, L95Q (D1) from wildtype. Makes it mesophilic +I-DmoI(site): + sequence: GCCTTGCCGGGTAAGTTCCGGCGCG diff --git a/parts/parts/openenzyme.yaml b/parts/parts/openenzyme.yaml new file mode 100644 index 0000000..dd4c225 --- /dev/null +++ b/parts/parts/openenzyme.yaml @@ -0,0 +1,30 @@ +# Ligases +HSlig4: + description: Ligase used in efficiently joining single-strand breaks in a double-strand polydeoxynucleotide in an ATP-dependent reaction. + sequence: MRLILPQLERERMAYGIKETMLAKLYIELLNLPRDGKDALKLLNYRTPTGTHGDAGDFAMIAYFVLKPRCLQKGSLTIQQVNDLLDSIASNNSAKRKDLIKKSLLQLITQSSALEQKWLIRMIIKDLKLGVSQQTIFSVFHNDAAELHNVTTDLEKVCRQLHDPSVGLSDISITLFSASKPMLAAIADIEHIEKDMKHQSFYIETKLDGERMQMHKDGDVYKYFSRNGYNYTDQFGASPTEGSLTPFIHNAFKADIQICILDGEMMAYNPNTQTFMQKGTKFDIKRMVEDSDLQTCYCVFDVLMVNNKKLGHETLRKRYEILSSIFTPIPGRIEIVQKTQAHTKNEVIDALNEAIDKREEGIMVKQPLSIYKPDKRGEGWLKIKPEYVSGLMDELDILIVGGYWGKGSRGGMMSHFLCAVAEKPPPGEKPSVFHTLSRVGSGCTMKELYDLGLKLAKYWKPFHRKAPPSSILCGTEKPEVYIEPCNSVIVQIKAAEIVPSDMYKTGCTLRFPRIEKIRDDKEWHECMTLDDLEQLRGKASGKLASKHLYIGGDDEPQEKKRKAAPKMKKVIGIIEHLKAPNLTNVNKISNIFEDVEFCVMSGTDSQPKPDLENRIAEFGGYIVQNPGPDTYCVIAGSENIRVKNIILSNKHDVVKPAWLLECFKTKSFVPWQPRFMIHMCPSTKEHFAREYDCYGDSYFIDTDLNQLKEVFSGIKNSNEQTPEEMASLIADLEYRYSWDCSPLSMFRRHTVYLDSYAVINDLSTKNEGTRLAIKALELRFHGAKVVSCLAEGVSHVIIGEDHSRVADFKAFRRTFKRKFKILKESWVTDSIDKCELQEENQYLI* +ECOligA: + description: Connects preferentially cohesive double-stranded DNA ends, active on blunt end DNA in the presence of Ficoll or polyethylene glycol. Requires Mg2+ and NAD+. Ligation when blunt end or RNA/ DNA ligation needs to be avoided. + sequence: MESIEQQLTELRTTLRHHEYLYHVMDAPEIPDAEYDRLMRELRELETKHPELITPDSPTQRVGAAPLAAFSQIRHEVPMLSLDNVFDEESFLAFNKRVQDRLKNNEKVTWCCELKLDGLAVSILYENGVLVSAATRGDGTTGEDITSNVRTIRAIPLKLHGENIPARLEVRGEVFLPQAGFEKINEDARRTGGKVFANPRNAAAGSLRQLDPRITAKRPLTFFCYGVGVLEGGELPDTHLGRLLQFKKWGLPVSDRVTLCESAEEVLAFYHKVEEDRPTLGFDIDGVVIKVNSLAQQEQLGFVARAPRWAVAFKFPAQEQMTFVRDVEFQVGRTGAITPVARLEPVHVAGVLVSNATLHNADEIERLGLRIGDKVVIRRAGDVIPQVVNVVLSERPEDTREVVFPTHCPVCGSDVERVEGEAVARCTGGLICGAQRKESLKHFVSRRAMDVDGMGDKIIDQLVEKEYVHTPADLFKLTAGKLTGLERMGPKSAQNVVNALEKAKETTFARFLYALGIREVGEATAAGLAAYFGTLEALEAASIEELQKVPDVGIVVASHVHNFFAEESNRNVISELLAEGVHWPAPIVINAEEIDSPFAGKTVVLTGSLSQMSRDDAKARLVELGAKVAGSVSKKTDLVIAGEAAGSKLAKAQELGIEVIDEAEMLRLLGS* +9N7lig: + description: 9 Degrees North DNA Ligase active at elevated temperatures (45 - 90 degrees C). + sequence: MKYSELAELYRRLEKTTLKTLKTKFVADFLKKTPDDLLEIVPYLILGKVFPDWDERELGVGEKLLIRAVSMATGVPEREIENSIKDTGDLGESVALALKKRKQKSFFSQPLTIKRVYSTFVKVAEASGEGSQDRKMKYLANLFMDAQPEEGKYIARTVLGTMRTGVAEGILRDAIAEAFKVKPELVERAYMLTSDFGYVAKVAKLEGNEGLSKVSIQIGKPIRPMLAQNAANVKEALIEMGGEAAFEIKYDGARVQVHRDGDRVIIYSRRLENVTRSIPEIVEAVKASLKPSKVIVEGELVAVGENGRPRPFQYVLRRFRRKYNIEEMIEKIPLELNLFDILYVDGESLIDTKFAERRKKLEESVEESDKIKLAEQLVTKKVEEAEEFYKRALELGHEGLMAKRLDSIYEPGNRGKKWLKIKPTMENLDLVIIGAEWGEGRRAHLLGSFLVAAYDPESGEFVPVGKVGSGFTDEDLVEFTKMLKPLIVREEGKFVEIEPKVVIEVTYQEIQKSPKYKSGFALRFPRYVALREDKSPEEADTIERVAQLYELQERFKAKR* +Pfulig: + description: DNA ligase that seals nicks in double-stranded DNA during DNA replication, DNA recombination and DNA repair. + sequence: MRYLELAQLYQKLEKTTMKLIKTRLVADFLKKVPDDHLEFIPYLILGEVFPEWDERELGVGEKLLIKAVAMATGIDAKEIEESVKDTGDLGESIALAVKKKKQKSFFSQPLTIKRVYQTLVKVAETTGEGSQDKKVKYLADLFMDAEPLEAKYLARTILGTMRTGVAEGLLRDAIAMAFHVKVELVERAYMLTSDFGYVAKIAKLEGNEGLAKVQVQLGKPIKPMLAQQAASIRDALLEMGGEAEFEIKYDGARVQVHKDGSKIIVYSRRLENVTRAIPEIVEALKEAIIPEKAIVEGELVAIGENGRPLPFQYVLRRFRRKHNIEEMMEKIPLELNLFDVLYVDGQSLIDTKFIDRRRTLEEIIKQNEKIKVAENLITKKVEEAEAFYKRALEMGHEGLMAKRLDAVYEPGNRGKKWLKIKPTMENLDLVIIGAEWGEGRRAHLFGSFILGAYDPETGEFLEVGKVGSGFTDDDLVEFTKMLKPLIIKEEGKRVWLQPKVVIEVTYQEIQKSPKYRSGFALRFPRFVALRDDKGPEDADTIERIAQLYELQEKMKGKVES* +THEAQlig: + description: Thermostable ligase for incorporation of phosphorylated oligonucleotides during PCR and Ligase Chain Reaction + sequence: MTLEAARRRVNELRDLIRYHNYLYYVLDAPEISDAEYDRLLRELKELEERFPELQSPDSPTEQVGARPLESTFRPVRHPTRMYSLDNAFSLDEVRAFEERIERALGRKGPFLYTVEHKVDGLSVNLYYEEGILVFGATRGDGETGEEVTQNLLTIRTIPRRLTGVPDRLEVRGEVYMPIEAFLRLNQELEEAGERIFKNPRNAAAGSLRQKDPRVTARRGLRATFYALGLGLEETGLKSQHDLLLWLRERGFPVEHGFTRALGAEGVEEVYQAWLKERRKLPFEADGVVVKLDDLALWRELGYTARAPRFALAYKFPAEEKETRLLSVAFQVGRTGRITPVGVLEPVFIEGSEVSRVTLHNESFIEELDVRIGDWVLVHKAGGVIPEVLRVLKERRTGEEKPILWPENCPECGHALIKEGKVHRCPNPLCPAKRFEAIRHYASRKAMDIQGLGEKLIEKLLEKGLVRDVADLYRLKKEDLVNLERMGEKSAENLLRQIEESKGRGLERLLYALGLPGVGEVLARNLALRFGHMDRLLEAGLEDLLEVEGVGELTARAILNTLKDPEFRDLVRRLKEAGVEMEAKEREGEALKGLTFVITGELSRPREEVKALLRRLGAKVTDSVSRKTGFLVVGENPGSKLEKARALGVPTLSEEELYRLIEERTGKDPRALTA* +T4gene30: + description: Ligase derived from T4 bacteriophage expressed in the early stage of lytic development. Has been implicated in T4 DNA synthesis and genetic recombination. May also play a role in T4 DNA repair. + sequence: MILKILNEIASIGSTKQKQAILEKNKDNELLKRVYRLTYSRGLQYYIKKWPKPGIATQSFGMLTLTDMLDFIEFTLATRKLTGNAAIEELTGYITDGKKDDVEVLRRVMMRDLECGASVSIANKVWPGLIPEQPQMLASSYDEKGINKNIKFPAFAQLKADGARCFAEVRGDELDDVRLLSRAGNEYLGLDLLKEELIKMTAEARQIHPEGVLIDGELVYHEQVKKEPEGLDFLFDAYPENSKAKEFAEVAESRTASNGIANKSLKGTISEKEAQCMKFQVWDYVPLVEIYSLPAFRLKYDVRFSKLEQMTSGYDKVILIENQVVNNLDEAKVIYKKYIDQGLEGIILKNIDGLWENARSKNLYKFKEVIDVDLKIVGIYPHRKDPTKAGGFILESECGKIKVNAGSGLKDKAGVKSHELDRTRIMENQNYYIGKILECECNGWLKSDGRTDYVKLFLPIAIRLREDKTKANTFEDVFGDFHEVTGL* +Tthlig: + description: Taq DNA Ligase + seqeunce: MTLEEARKRVNELRDLIRYHNYRYYVLADPEISDAEYDRLLRELKELEERFPELKSPDSPTLQVGARPLEATFRPVRHPTRMYSLDNAFNLDELKAFEERIERALGRKGPFAYTVEHKVDGLSVNLYYEEGVLVYGATRGDGEVGEEVTQNLLTIPTIPRRLKGVPERLEVRGEVYMPIEAFLRLNEELEERGERIFKNPRNAAAGSLRQKDPRITAKRGLRATFYALGLGLEEVEREGVATQFALLHWLKEKGFPVEHGYARAVGAEGVEAVYQDWLKKRRALPFEADGVVVKLDELALWRELGYTARAPRFAIAYKFPAEEKETRLLDVVFQVGRTGRVTPVGILEPVFLEGSEVSRVTLHNESYIEELDIRIGDWVLVHKAGGVIPEVLRVLKERRTGEERPIRWPETCPECGHRLLKEGKVHRCPNPLCPAKRFEAIRHFASRKAMDIQGLGEKLIERLLEKGLVKDVADLYRLRKEDLVGLERMGEKSAQNLLRQIEESKKRGLERLLYALGLPGVGEVLARNLAARFGNMDRLLEASLEELLEVEEVGELTARAILETLKDPAFRDLVRRLKEAGVEMEAKEKGGEALKGLTFVITGELSRPREEVKALLRRLGAKVTDSVSRKTSYLVVGENPGSKLEKARALGVPTLTEEELYRLLEARTGKKAEELV* +HSlig3: + description: Repairs single strand breaks in DNA efficiently. Unable to perform either blunt-end joining or AMP- dependent relaxation of supercoiled DNA (Elder, R.H. et al . , Bur. J. Biochem.. 203:53-58 (1992)) + sequence: MSLAFKIFFPQTLRALSRKELCLFRKHHWRDVRQFSQWSETDLLHGHPLFLRRKPVLSFQGSHLRSRATYLVFLPGLHVGLCSGPCEMAEQRFCVDYAKRGTAGCKKCKEKIVKGVCRIGKVVPNPFSESGGDMKEWYHIKCMFEKLERARATTKKIEDLTELEGWEELEDNEKEQITQHIADLSSKAAGTPKKKAVVQAKLTTTGQVTSPVKGASFVTSTNPRKFSGFSAKPNNSGEAPSSPTPKRSLSSSKCDPRHKDCLLREFRKLCAMVADNPSYNTKTQIIQDFLRKGSAGDGFHGDVYLTVKLLLPGVIKTVYNLNDKQIVKLFSRIFNCNPDDMARDLEQGDVSETIRVFFEQSKSFPPAAKSLLTIQEVDEFLLRLSKLTKEDEQQQALQDIASRCTANDLKCIIRLIKHDLKMNSGAKHVLDALDPNAYEAFKASRNLQDVVERVLHNAQEVEKEPGQRRALSVQASLMTPVQPMLAEACKSVEYAMKKCPNGMFSEIKYDGERVQVHKNGDHFSYFSRSLKPVLPHKVAHFKDYIPQAFPGGHSMILDSEVLLIDNKTGKPLPFGTLGVHKKAAFQDANVCLFVFDCIYFNDVSLMDRPLCERRKFLHDNMVEIPNRIMFSEMKRVTKALDLADMITRVIQEGLEGLVLKDVKGTYEPGKRHWLKVKKDYLNEGAMADTADLVVLGAFYGQGSKGGMMSIFLMGCYDPGSQKWCTVTKCAGGHDDATLARLQNELDMVKISKDPSKIPSWLKVNKIYYPDFIVPDPKKAAVWEITGAEFSKSEAHTADGISIRFPRCTRIRDDKDWKSATNLPQLKELYQLSKEKADFTVVAGDEGSSTTGGSSEENKGPSGSAVSRKAPSKPSASTKKAEGKLSNSNSKDGNMQTAKPSAMKVGEKLATKSSPVKVGEKRKAADETLCQTKVLLDIFTGVRLYLPPSTPDFSRLRRYFVAFDGDLVQEFDMTSATHVLGSRDKNPAAQQVSPEWIWACIRKRRLVAPC* + +# DNA polymerase +# Reverse Transcriptase +# RNA polymerase +# Other diff --git a/parts/parts/proteins/HSlig4.md b/parts/parts/proteins/HSlig4.md new file mode 100644 index 0000000..f1e73b1 --- /dev/null +++ b/parts/parts/proteins/HSlig4.md @@ -0,0 +1,6 @@ +--- +name: HSlig4 +short_description: Ligase used in efficiently joining single-strand breaks in a double-strand polydeoxynucleotide in an ATP-dependent reaction. +sequence: MRLILPQLERERMAYGIKETMLAKLYIELLNLPRDGKDALKLLNYRTPTGTHGDAGDFAMIAYFVLKPRCLQKGSLTIQQVNDLLDSIASNNSAKRKDLIKKSLLQLITQSSALEQKWLIRMIIKDLKLGVSQQTIFSVFHNDAAELHNVTTDLEKVCRQLHDPSVGLSDISITLFSASKPMLAAIADIEHIEKDMKHQSFYIETKLDGERMQMHKDGDVYKYFSRNGYNYTDQFGASPTEGSLTPFIHNAFKADIQICILDGEMMAYNPNTQTFMQKGTKFDIKRMVEDSDLQTCYCVFDVLMVNNKKLGHETLRKRYEILSSIFTPIPGRIEIVQKTQAHTKNEVIDALNEAIDKREEGIMVKQPLSIYKPDKRGEGWLKIKPEYVSGLMDELDILIVGGYWGKGSRGGMMSHFLCAVAEKPPPGEKPSVFHTLSRVGSGCTMKELYDLGLKLAKYWKPFHRKAPPSSILCGTEKPEVYIEPCNSVIVQIKAAEIVPSDMYKTGCTLRFPRIEKIRDDKEWHECMTLDDLEQLRGKASGKLASKHLYIGGDDEPQEKKRKAAPKMKKVIGIIEHLKAPNLTNVNKISNIFEDVEFCVMSGTDSQPKPDLENRIAEFGGYIVQNPGPDTYCVIAGSENIRVKNIILSNKHDVVKPAWLLECFKTKSFVPWQPRFMIHMCPSTKEHFAREYDCYGDSYFIDTDLNQLKEVFSGIKNSNEQTPEEMASLIADLEYRYSWDCSPLSMFRRHTVYLDSYAVINDLSTKNEGTRLAIKALELRFHGAKVVSCLAEGVSHVIIGEDHSRVADFKAFRRTFKRKFKILKESWVTDSIDKCELQEENQYLI +--- +# HSlig4 From 183cd20ec5f0fe9b4746a40ed57c7d9697a3ca12 Mon Sep 17 00:00:00 2001 From: Keoni Gandall Date: Sun, 20 Oct 2024 23:17:16 -0700 Subject: [PATCH 8/9] finish openenzyme --- parts/parts/openenzyme.yaml | 139 +++++++++++++++++++++++++++++++++++- 1 file changed, 138 insertions(+), 1 deletion(-) diff --git a/parts/parts/openenzyme.yaml b/parts/parts/openenzyme.yaml index dd4c225..0af8376 100644 --- a/parts/parts/openenzyme.yaml +++ b/parts/parts/openenzyme.yaml @@ -23,8 +23,145 @@ Tthlig: HSlig3: description: Repairs single strand breaks in DNA efficiently. Unable to perform either blunt-end joining or AMP- dependent relaxation of supercoiled DNA (Elder, R.H. et al . , Bur. J. Biochem.. 203:53-58 (1992)) sequence: MSLAFKIFFPQTLRALSRKELCLFRKHHWRDVRQFSQWSETDLLHGHPLFLRRKPVLSFQGSHLRSRATYLVFLPGLHVGLCSGPCEMAEQRFCVDYAKRGTAGCKKCKEKIVKGVCRIGKVVPNPFSESGGDMKEWYHIKCMFEKLERARATTKKIEDLTELEGWEELEDNEKEQITQHIADLSSKAAGTPKKKAVVQAKLTTTGQVTSPVKGASFVTSTNPRKFSGFSAKPNNSGEAPSSPTPKRSLSSSKCDPRHKDCLLREFRKLCAMVADNPSYNTKTQIIQDFLRKGSAGDGFHGDVYLTVKLLLPGVIKTVYNLNDKQIVKLFSRIFNCNPDDMARDLEQGDVSETIRVFFEQSKSFPPAAKSLLTIQEVDEFLLRLSKLTKEDEQQQALQDIASRCTANDLKCIIRLIKHDLKMNSGAKHVLDALDPNAYEAFKASRNLQDVVERVLHNAQEVEKEPGQRRALSVQASLMTPVQPMLAEACKSVEYAMKKCPNGMFSEIKYDGERVQVHKNGDHFSYFSRSLKPVLPHKVAHFKDYIPQAFPGGHSMILDSEVLLIDNKTGKPLPFGTLGVHKKAAFQDANVCLFVFDCIYFNDVSLMDRPLCERRKFLHDNMVEIPNRIMFSEMKRVTKALDLADMITRVIQEGLEGLVLKDVKGTYEPGKRHWLKVKKDYLNEGAMADTADLVVLGAFYGQGSKGGMMSIFLMGCYDPGSQKWCTVTKCAGGHDDATLARLQNELDMVKISKDPSKIPSWLKVNKIYYPDFIVPDPKKAAVWEITGAEFSKSEAHTADGISIRFPRCTRIRDDKDWKSATNLPQLKELYQLSKEKADFTVVAGDEGSSTTGGSSEENKGPSGSAVSRKAPSKPSASTKKAEGKLSNSNSKDGNMQTAKPSAMKVGEKLATKSSPVKVGEKRKAADETLCQTKVLLDIFTGVRLYLPPSTPDFSRLRRYFVAFDGDLVQEFDMTSATHVLGSRDKNPAAQQVSPEWIWACIRKRRLVAPC* - # DNA polymerase +phi29pol: + description: DNA polymerase responsible for protein-primed viral DNA replication by strand displacement with high processivity and fidelity. Possesses three enzymatic activities - DNA synthesis (polymerase), primer terminal protein (TP) deoxynucleotidylation, and 3' to 5' exonuclease activity. + sequence: MKHMPRKMYSCDFETTTKVEDCRVWAYGYMNIEDHSEYKIGNSLDEFMAWVLKVQADLYFHNLKFDGAFIINWLERNGFKWSADGLPNTYNTIISRMGQWYMIDICLGYKGKRKIHTVIYDSLKKLPFPVKKIAKDFKLTVLKGDIDYHKERPVGYKITPEEYAYIKNDIQIIAERLLIQFKQGLDRMTAGSDSLKGFKDIITTKKFKKVFPTLSLGLDKEVRYAYRGGFTWLNDRFKEKEIGEGMVFDVNSLYPAQMYSRLLPYGEPIVFEGKYVWDEDYPLHIQHIRCEFELKEGYIPTIQIKRSRFYKGNEYLKSSGGEIADLWLSNVDLELMKEHYDLYNVEYISGLKFKATTGLFKDFIDKWTYIKTTSEGAIKQLAKLMLNSLYGKFASNPDVTGKVPYLKENGALGFRLGEEETKDPVYTPMGVFITAWARYTTITAAQACYDRIIYCDTDSIHLTGTEIPDVIKDIVDPKKLGYWAHESTFKRVKYLRQKTYIQDIYMKEVDGKLVEGSPDDYTDIKFSVKCAGMTDKIKKEVTFENFKVGFSRKMKPKPVQVPGGVVLVDDTFTIK* +Pwopol: + description: DNA polymerase; additionally exhibits 3' to 5' exonuclease activity. + sequence: MILDVDYITEEGKPVIRLFKKENGKFKIEHDRTFRPYIYALLRDDSKIEEVKKITGERHGKIVRIVDVEKVEKKFLGKPITVWKLYLEHPQDVPTIREKVREHPAVVDIFEYDIPFAKRYLIDKGLIPMEGEEELKILAFDIETLYHEGEEFGKGPIIMISYADENEAKVITWKNIDLPYVEVVSSEREMIKRFLRIIREKDPDIIVTYNGDSFDFPYLAKRAEKLGIKLTIGRDGSEPKMQRIGDMTAVEVKGRIHFDLYHVITRTINLPTYTLEAVYEAIFGKPKEKVYADEIAKAWESGENLERVAKYSMEDAKATYELGKEFLPMEIQLSRLVGQPLWDVSRSSTGNLVEWFLLRKAYERNEVAPNKPSEEEYQRRLRESYTGGFVKEPEKGLWENIVYLDFRALYPSIIITHNVSPDTLNLEGCKNYDIAPQVGHKFCKDIPGFIPSLLGHLLEERQKIKTKMKETQDPIEKILLDYRQKAIKLLANSFYGYYGYAKARWYCKECAESVTAWGRKYIELVWKELEEKFGFKVLYIDTDGLYATIPGGESEEIKKKALEFVKYINSKLPGLLELEYEGFYKRGFFVTKKRYAVIDEEGKVITRGLEIVRRDWSEIAKETQARVLETILKHGDVEEAVRIVKEVIQKLANYEIPPEKLAIYEQITRPLHEYKAIGPHVAVAKKLAAKGVKIKPGMVIGYIVLRGDGPISNRAILAEEYDPKKHKYDAEYYIENQVLPAVLRILEGFGYRKEDLRYQKTRQVGLTSWLNIKKS* +GBDpolexo: + description: Robust and extremely thermostable polymerase with a half-life of 23 hours at 95 degrees C; offers 5x higher fidelity than Taq and robust performance. Ideal for GC-rich or looped sequences. Lacks exonuclease activity. Comparable to Deep Vent (exo-) DNA Polymerase at NEB + sequence: MIIDADYITEDGKPIIRIFKKEKGEFKVEYDRTFRPYIYALLKDDSAIDEVKKITAERHGKIVRITEVEKVQKKFLGRPIEVWKLYLEHPQDVPAIREKIREHPAVVDIFEYDIPFAKRYLIDKGLTPMEGNEELTFLAVAIATLYHEGEEFGKGPIIMISYADEEGAKVITWKSIDLPYVEVVSSEREMIKRLVKVIREKDPDVIITYNGDNFDFPYLLKRAEKLGIKLPLGRDNSEPKMQRMGDSLAVEIKGRIHFDLFPAIRRTINLPTYTLETVYEVIFGKSKEKVYAHEIAEAWETGKGLERVAKYSMEDAKVTSELGKEFFPMEAQLARLVGHPVWDVSRSSTGNLVEWFLLTKAYERNELAPNKPDEREYERRLRESYEGGYVNEPEKGLWEGIVSLDFRSLYPSIIITHNVSPDTLNRENCKEYDVAPQVGHRFCKDFPGFIPSLLGNLLEERQKIKKRMKESKDPVEKKLLDYRQRAIKILANSYYGYYGYAKARWYCKECAESVTAWGRQYIDLVRRELESRGFKVLYIDTDGLYATIPGAKHEEIKEKALKFVEYINSKLPGLLELEYEGFYARGFFVTKKKYALIDEEGKIVTRGLEIVRRDWSEIAKETQAKVLEAILKHGNVDEAVKIVKEVTEKLSKYEIPPEKLVIYEQITRPLSEYKAIGPHVAVAKRLAAKGVKVKPGMVIGYIVLKGDGPISKRAIAIEEFDPKKHKYDAEYYIENQVLPAVERILRAFGYRKEDLRYQKTKQVGLGAWLKF* +P45: + description: Significantly improves maximum amplicon size and PCR yields for Family B polymerases like Pfu or Phusion. Comparable to ArchaeMaxx/PfuTurbo. + sequence: MLHHVKLIYATKSRKLVGKKIVLAIPGSIAAVECVKLARELIRHGAEVHAVMSEAATKIIHPYAMEFATGNPVITEITGFIEHVELAGEHENKADLILVCPATANTISKIACGIDDTPVTTVVTTAFPHIPIMIAPAMHETMYRHPIVRENIERLKKLGVEFIGPRIEEGKAKVASIDEIVYRVIKKLHKKTLEGKRVLVTAGATREYIDPIRFITNASSGKMGVALAEEADFRGAEVTLIRTKGSVKSFVENQIEVETVEEMLSAIENELRSKKYDVVIMAAAVSDFRPKIKAEGKIKSDRSITIELVPNPKIIDRIKEIQPNVFLVGFKAETSKEKLIEEGKRQIERAKADLVVGNTLEAFGSEENQVVLIGRDFTKELPKMKKRELAERIWDEIEKLLS* +T5gene122: + description: Possesses two enzymatic activities - DNA synthesis (polymerase) and exonucleolytic activity that degrades ssDNA in the 3'-5' direction for proofreading purposes. + sequence: MKIAVVDKALNNTRYDKHFQLYGEEVDVFHMCNEKLSGRLLKKHITIGTPENPFDPNDYDFVILVGAEPFLYFAGKKGIGDYTGKRVEYNGYANWIASISPAQLHFKPEMKPVFDATVENIHDIINGREKIAKAGDYRPITDPDEAEEYIKMVYNMVIGPVAFDSETSALYCRDGYLLGVSISHQEYQGVYIDSDCLTEVAVYYLQKILDSENHTIVFHNLKFDMHFYKYHLGLTFDKAHKERRLHDTMLQHYVLDERRGTHGLKSLAMKYTDMGDYDFELDKFKDDYCKAHKIKKEDFTYDLIPFDIMWPYAAKDTDATIRLHNFFLPKIEKNEKLCSLYYDVLMPGCVFLQRVEDRGVPISIDRLKEAQYQLTHNLNKAREKLYTYPEVKQLEQDQNEAFNPNSVKQLRVLLFDYVGLTPTGKLTDTGADSTDAEALNELATQHPIAKTLLEIRKLTKLISTYVEKILLSIDADGCIRTGFHEHMTTSGRLSSSGKLNLQQLPRDESIIKGCVVAPPGYRVIAWDLTTAEVYYAAVLSGDRNMQQVFINMRNEPDKYPDFHSNIAHMVFKLQCEPRDVKKLFPALRQAAKAITFGILYGSGPAKVAHSVNEALLEQAAKTGEPFVECTVADAKEYIETYFGQFPQLKRWIDKCHDQIKNHGFIYSHFGRKRRLHNIHSEDRGVQGEEIRSGFNAIIQSASSDSLLLGAVDADNEIISLGLEQEMKIVMLVHDSVVAIVREDLIDQYNEILIRNIQKDRGISIPGCPIGIDSDSEAGGSRDYSCGKMKKQHPSIACIDDDEYTRYVKGVLLDAEFEYKKLAAMDKEHPDHSKYKDDKFIAVCKDLDNVKRILGA* +BstpolLF: + description: Fragment retains 5'-3' polymerase activity from full length Bst DNA Polymerase, while lacking 5'-3' exonuclease activity. Suitable for applications requiring thermophilic strand displacement. + sequence: MTDEGEKPLAGMDFAIADSVTDEMLADKAALVVEVVGDNYHHAPIVGIALANERGRFFLRPETALADPKFLAWLGDETKKKTMFDSKRAAVALKWKGIELRGVVFDLLLAAYLLDPAQAAGDVAAVAKMHQYEAVRSDEAVYGKGAKRTVPDEPTLAEHLARKAAAIWALEEPLMDELRRNEQDRLLTELEQPLAGILANMEFTGVKVDTKRLEQMGAELTEQLQAVERRIYELAGQEFNINSPKQLGTVLFDKLQLPVLKKTKTGYSTSADVLEKLAPHHEIVEHILHYRQLGKLQSTYIEGLLKVVHPVTGKVHTMFNQALTQTGRLSSVEPNLQNIPIRLEEGRKIRQAFVPSEPDWLIFAADYSQIELRVLAHIAEDDNLIEAFRRGLDIHTKTAMDIFHVSEEDVTANMRRQAKAVNFGIVYGISDYGLAQNLNITRKEAAEFIERYFASFPGVKQYMDNIVQEAKQKGYVTTLLHRRRYLPDITSRNFNVRSFAERTAMNTPIQGSAADIIKKAMIDLSVRLREERLQARLLLQVHDELILEAPKEEIERLCRLVPEVMEQAVALRVPLKVDYHYGPTWYDAK* +THEAQpolA: + description: DNA polymerase used in routine PCR method because of its high thermostability, standard testing, screening and output testing. + sequence: MRGMLPLFEPKGRVLLVDGHHLAYRTFHALKGLTTSRGEPVQAVYGFAKSLLKALKEDGDAVIVVFDAKAPSFRHEAYGGYKAGRAPTPEDFPRQLALIKELVDLLGLARLEVPGYEADDVLASLAKKAEKEGYEVRILTADKDLYQLLSDRIHVLHPEGYLITPAWLWEKYGLRPDQWADYRALTGDESDNLPGVKGIGEKTARKLLEEWGSLEALLKNLDRLKPAIREKILAHMDDLKLSWDLAKVRTDLPLEVDFAKRREPDRERLRAFLERLEFGSLLHEFGLLESPKALEEAPWPPPEGAFVGFVLSRKEPMWADLLALAAARGGRVHRAPEPYKALRDLKEARGLLAKDLSVLALREGLGLPPGDDPMLLAYLLDPSNTTPEGVARRYGGEWTEEAGERAALSERLFANLWGRLEGEERLLWLYREVERPLSAVLAHMEATGVRLDVAYLRALSLEVAEEIARLEAEVFRLAGHPFNLNSRDQLERVLFDELGLPAIGKTEKTGKRSTSAAVLEALREAHPIVEKILQYRELTKLKSTYIDPLPDLIHPRTGRLHTRFNQTATATGRLSSSDPNLQNIPVRTPLGQRIRRAFIAEEGWLLVALDYSQIELRVLAHLSGDENLIRVFQEGRDIHTETASWMFGVPREAVDPLMRRAAKTINFGVLYGMSAHRLSQELAIPYEEAQAFIERYFQSFPKVRAWIEKTLEEGRRRGYVETLFGRRRYVPDLEARVKSVREAAERMAFNMPVQGTAADLMKLAMVKLFPRLEEMGARMLLQVHDELVLEAPKERAEAVARLAKEVMEGVYPLAVPLEVEVGIGEDWLSAKE* +BsupolLF: + description: Bsu DNA Polymerase I, Large Fragment retains the 5'-3' polymerase activity of the Bacillus subtilis DNA polymerase I, but lacks the 5'-3' exonuclease domain. This large fragment naturally lacks 3'-5' exonuclease activity. Applications include random primer labeling, second strand cDNA synthesis, single dA tailing, and strand displacement DNA synthesis. + sequence: MGVGEKTAIKLLKQFDSVEKLLESIDEVSGKKLKEKLEEFKDQALMSKELATIMTDAPIEVSVSGLEYQGFNREQVIAIFKDLGFNTLLERLGEDSAEAEQDQSLEDINVKTVTDVTSDILVSPSAFVVEQIGDNYHEEPILGFSIVNETGAYFIPKDIAVESEVFKEWVENDEQKKWVFDSKRAVVALRWQGIELKGAEFDTLLAAYIINPGNSYDDVASVAKDYGLHIVSSDESVYGKGAKRAVPSEDVLSEHLGRKALAIQSLREKLVQELENNDQLELFEELEMPLALILGEMESTGVKVDVDRLKRMGEELGAKLKEYEEKIHEIAGEPFNINSPKQLGVILFEKIGLPVVKKTKTGYSTSADVLEKLADKHDIVDYILQYRQIGKLQSTYIEGLLKVTRPDSHKVHTRFNQALTQTGRLSSTDPNLQNIPIRLEEGRKIRQAFVPSEKDWLIFAADYSQIELRVLAHISKDENLIEAFTNDMDIHTKTAMDVFHVAKDEVTSAMRRQAKAVNFGIVYGISDYGLSQNLGITRKEAGAFIDRYLESFQGVKAYMEDSVQEAKQKGYVTTLMHRRRYIPELTSRNFNIRSFAERTAMNTPIQGSAADIIKKAMIDMAAKLKEKQLKARLLLQVHDELIFEAPKEEIEILEKLVPEVMEHALALDVPLKVDFASGPSWYDAK* +Bstpol: + description: Full length DNA polymerase from Bacillus stearothermophilus. + sequence: MKNKLVLIDGNSVAYRAFFALPLLHNDKGIHTNAVYGFTMMLNKILAEEQPTHILVAFDAGKTTFRHETFQDYKGGRQQTPPELSEQFPLLRELLKAYRIPAYELDHYEADDIIGTMAARAEREGFAVKVISGDRDLTQLASPQVTVEITKKGITDIESYTPETVVEKYGLTPEQIVDLKGLMGDKSDNIPGVPGIGEKTAVKLLKQFGTVENVLASIDEIKGEKLKENLRQYRDLALLSKQLAAICRDAPVELTLDDIVYKGEDREKVVALFQELGFQSFLDKMAVQTDEGEKPLAGMDFAIADSVTDEMLADKAALVVEVVGDNYHHAPIVGIALANERGRFFLRPETALADPKFLAWLGDETKKKTMFDSKRAAVALKWKGIELRGVVFDLLLAAYLLDPAQAAGDVAAVAKMHQYEAVRSDEAVYGKGAKRTVPDEPTLAEHLARKAAAIWALEEPLMDELRRNEQDRLLTELEQPLAGILANMEFTGVKVDTKRLEQMGAELTEQLQAVERRIYELAGQEFNINSPKQLGTVLFDKLQLPVLKKTKTGYSTSADVLEKLAPHHEIVEHILHYRQLGKLQSTYIEGLLKVVHPVTGKVHTMFNQALTQTGRLSSVEPNLQNIPIRLEEGRKIRQAFVPSEPDWLIFAADYSQIELRVLAHIAEDDNLIEAFRRGLDIHTKTAMDIFHVSEEDVTANMRRQAKAVNFGIVYGISDYGLAQNLNITRKEAAEFIERYFASFPGVKQYMDNIVQEAKQKGYVTTLLHRRRYLPDITSRNFNVRSFAERTAMNTPIQGSAADIIKKAMIDLSVRLREERLQARLLLQVHDELILEAPKEEIERLCRLVPEVMEQAVALRVPLKVDYHYGPTWYDAK* +GBDpol: + description: Robust and extremely thermostable polymerase with 5x higher fidelity than Taq. Ideal for GC-rich or looped sequences. + sequence: MIIDADYITEDGKPIIRIFKKEKGEFKVEYDRTFRPYIYALLKDDSAIDEVKKITAERHGKIVRITEVEKVQKKFLGRPIEVWKLYLEHPQDVPAIREKIREHPAVVDIFEYDIPFAKRYLIDKGLTPMEGNEELTFLAVDIETLYHEGEEFGKGPIIMISYADEEGAKVITWKSIDLPYVEVVSSEREMIKRLVKVIREKDPDVIITYNGDNFDFPYLLKRAEKLGIKLPLGRDNSEPKMQRMGDSLAVEIKGRIHFDLFPAIRRTINLPTYTLETVYEVIFGKSKEKVYAHEIAEAWETGKGLERVAKYSMEDAKVTSELGKEFFPMEAQLARLVGHPVWDVSRSSTGNLVEWFLLTKAYERNELAPNKPDEREYERRLRESYEGGYVNEPEKGLWEGIVSLDFRSLYPSIIITHNVSPDTLNRENCKEYDVAPQVGHRFCKDFPGFIPSLLGNLLEERQKIKKRMKESKDPVEKKLLDYRQRAIKILANSYYGYYGYAKARWYCKECAESVTAWGRQYIDLVRRELESRGFKVLYIDTDGLYATIPGAKHEEIKEKALKFVEYINSKLPGLLELEYEGFYARGFFVTKKKYALIDEEGKIVTRGLEIVRRDWSEIAKETQAKVLEAILKHGNVDEAVKIVKEVTEKLSKYEIPPEKLVIYEQITRPLSEYKAIGPHVAVAKRLAAKGVKVKPGMVIGYIVLKGDGPISKRAIAIEEFDPKKHKYDAEYYIENQVLPAVERILRAFGYRKEDLRYQKTKQVGLGAWLKF* +TthpolA: + description: DNA Polymerase whose activity is resistant to prolonged incubations at high temperatures (95 degrees C). + sequence: MEAMLPLFEPKGRVLLVDGHHLAYRTFFALKGLTTSRGEPVQAVYGFAKSLLKALKEDGYKAVFVVFDAKAPSFRHEAYEAYKAGRAPTPEDFPRQLALIKELVDLLGFTRLEVPGYEADDVLATLAKKAEKEGYEVRILTADRDLYQLVSDRVAVLHPEGHLITPEWLWEKYGLRPEQWVDFRALVGDPSDNLPGVKGIGEKTALKLLKEWGSLENLLKNLDRVKPENVREKIKAHLEDLRLSLELSRVRTDLPLEVDLAQGREPDREGLRAFLERLEFGSLLHEFGLLEAPAPLEEAPWPPPEGAFVGFVLSRPEPMWAELKALAACRDGRVHRAADPLAGLKDLKEVRGLLAKDLAVLASREGLDLVPGDDPMLLAYLLDPSNTTPEGVARRYGGEWTEDAAHRALLSERLHRNLLKRLEGEEKLLWLYHEVEKPLSRVLAHMEATGVRRDVAYLQALSLELAEEIRRLEEEVFRLAGHPFNLNSRDQLERVLFDELRLPALGKTQKTGKRSTSAAVLEALREAHPIVEKILQHRELTKLKNTYVDPLPSLVHPRTGRLHTRFNQTATATGRLSSSDPNLQNIPVRTPLGQRIRRAFVAEAGWALVALDYSQIELRVLAHLSGDENLIRVFQEGKDIHTQTASWMFGVPPEAVDPLMRRAAKTVNFGVLYGMSAHRLSQELAIPYEEAVAFIERYFQSFPKVRAWIEKTLEEGRKRGYVETLFGRRRYVPDLNARVKSVREAAERMAFNMPVQGTAADLMKLAMVKLFPRLREMGARMLLQVHDELLLEAPQARAEEVAALAKEAMEKAYPLAVPLEVEVGMGEDWLSAKG* +Tlipolexo: + description: DNA polymerase; lacks 3' to 5' exonuclease activity. + sequence: MILDTDYITKDGKPIIRIFKKENGEFKIELDPHFQPYIYALLKDDSAIEEIKAIKGERHGKTVRVLDAVKVRKKFLGREVEVWKLIFEHPQDVPAMRGKIREHPAVVDIYEYDIPFAKRYLIDKGLIPMEGDEELKLLAFAIATFYHEGDEFGKGEIIMISYADEEEARVITWKNIDLPYVDVVSNEREMIKRFVQVVKEKDPDVIITYNGDNFDLPYLIKRAEKLGVRLVLGRDKEHPEPKIQRMGDSFAVEIKGRIHFDLFPVVRRTINLPTYTLEAVYEAVLGKTKSKLGAEEIAAIWETEESMKKLAQYSMEDARATYELGKEFFPMEAELAKLIGQSVWDVSRSSTGNLVEWYLLRVAYARNELAPNKPDEEEYKRRLRTTYLGGYVKEPEKGLWENIIYLDFRSLYPSIIVTHNVSPDTLEKEGCKNYDVAPIVGYRFCKDFPGFIPSILGDLIAMRQDIKKKMKSTIDPIEKKMLDYRQRAIKLLANSYYGYMGYPKARWYSKECAESVTAWGRHYIEMTIREIEEKFGFKVLYADTDGFYATIPGEKPELIKKKAKEFLNYINSKLPGLLELEYEGFYLRGFFVTKKRYAVIDEEGRITTRGLEVVRRDWSEIAKETQAKVLEAILKEGSVEKAVEVVRDVVEKIAKYRVPLEKLVIHEQITRDLKDYKAIGPHVAIAKRLAARGIKVKPGTIISYIVLKGSGKISDRVILLTEYDPRKHKYDPDYYIENQVLPAVLRILEAFGYRKEDLRYQSSKQTGLDAWLKR* +T4gene43: + description: Polymerase that replicates viral genomic DNA. + sequence: MKEFYISIETVGNNIVERYIDENGKERTREVEYLPTMFRHCKEESKYKDIYGKNCAPQKFPSMKDARDWMKRMEDIGLEALGMNDFKLAYISDTYGSEIVYDRKFVRVANCDIEVTGDKFPDPMKAEYEIDAITHYDSIDDRFYVFDLLNSMYGSVSKWDAKLAAKLDCEGGDEVPQEILDRVIYMPFDNERDMLMEYINLWEQKRPAIFTGWNIEGFDVPYIMNRVKMILGERSMKRFSPIGRVKSKLIQNMYGSKEIYSIDGVSILDYLDLYKKFAFTNLPSFSLESVAQHETKKGKLPYDGPINKLRETNHQRYISYNIIDVESVQAIDKIRGFIDLVLSMSYYAKMPFSGVMSPIKTWDAIIFNSLKGEHKVIPQQGSHVKQSFPGAFVFEPKPIARRYIMSFDLTSLYPSIIRQVNISPETIRGQFKVHPIHEYIAGTAPKPSDEYSCSPNGWMYDKHQEGIIPKEIAKVFFQRKDWKKKMFAEEMNAEAIKKIIMKGAGSCSTKPEVERYVKFSDDFLNELSNYTESVLNSLIEECEKAATLANTNQLNRKILINSLYGALGNIHFRYYDLRNATAITIFGQVGIQWIARKINEYLNKVCGTNDEDFIAAGDTDSVYVCVDKVIEKVGLDRFKEQNDLVEFMNQFGKKKMEPMIDVAYRELCDYMNNREHLMHMDREAISCPPLGSKGVGGFWKAKKRYALNVYDMEDKRFAEPHLKIMGMETQQSSTPKAVQEALEESIRRILQEGEESVQEYYKNFEKEYRQLDYKVIAEVKTANDIAKYDDKGWPGFKCPFHIRGVLTYRRAVSGLGVAPILDGNKVMVLPLREGNPFGDKCIAWPSGTELPKEIRSDVLSWIDHSTLFQKSFVKPLAGMCESAGMDYEEKASLDFLFG* +K12polLF: + description: DNA pol fragment that retains 5'-3' polymerase activity and 3'-5' exonuclease activity but loses 5'-3' exonuclease activity. + sequence: MVISYDNYVTILDEETLKAWIAKLEKAPVFAFDTETDSLDNISANLVGLSFAIEPGVAAYIPVAHDYLDAPDQISRERALELLKPLLEDEKALKVGQNLKYDRGILANYGIELRGIAFDTMLESYILNSVAGRHDMDSLAERWLKHKTITFEEIAGKGKNQLTFNQIALEEAGRYAAEDADVTLQLHLKMWPDLQKHKGPLNVFENIEMPLVPVLSRIERNGVKIDPKVLHNHSEELTLRLAELEKKAHEIAGEEFNLSSTKQLQTILFEKQGIKPLKKTPGGAPSTSEEVLEELALDYPLPKVILEYRGLAKLKSTYTDKLPLMINPKTGRVHTSYHQAVTATGRLSSTDPNLQNIPVRNEEGRRIRQAFIAPEDYVIVSADYSQIELRIMAHLSRDKGLLTAFAEGKDIHRATAAEVFGLPLETVTSEQRRSAKAINFGLIYGMSAFGLARQLNIPRKEAQKYMDLYFERYPGVLEYMERTRAQAKEQGYVETLDGRRLYLPDIKSSNGARRAAAERAAINAPMQGTAADIIKRAMIAVDAWLQAEQPRVRMIMQVHDELVFEVHKDDVDAVAKQIHQLMENCTRLDVPLLVEVGSGENWDQAH* +K12polLFexo: + description: DNA pol fragment used in flourescent labelling for microarray, dA and dT tailing, and ligating DNA adapters to DNA fragments. + sequence: MGPLNVFENIEMPLVPVLSRIERNGVKIDPKVLHNHSEELTLRLAELEKKAHEIAGEEFNLSSTKQLQTILFEKQGIKPLKKTPGGAPSTSEEVLEELALDYPLPKVILEYRGLAKLRSTYTDKLPLMINPKTGRVHTSYHQAVTATGRLSSTDPNLQNIPVRNEEGRRIRQAFIAPEDYVIVSADYSQIELRIMAHLSRDKGLLTAFAEGKDIHRATAAEVFGLPLETVTSEQRRSAKAINFGLIYGMSAFGLARQLNIPRKEAQKYMDLYFERYPGVLEYMERTRAQAKEQGYVETLDGRRLYLPDIKSSNGARRAAAERAAINAPMQGTAADIIKRAMIAVDAWLQAEQPRVRMIMQVHDELVFEVHKDDVDAVAKQIHQLMENCTRLDVPLLVEVGSGENWDQAH* +9N7polA: + description: Thermophilic DNA polymerase; genetically engineered to have a decreased 3'-5' proofreading exonuclease activity + sequence: MILDTDYITENGKPVIRVFKKENGEFKIEYDRTFEPYFYALLKDDSAIEDVKKVTAKRHGTVVKVKRAEKVQKKFLGRPIEVWKLYFNHPQDVPAIRDRIRAHPAVVDIYEYDIPFAKRYLIDKGLIPMEGDEELTMLAFDIETLYHEGEEFGTGPILMISYADGSEARVITWKKIDLPYVDVVSTEKEMIKRFLRVVREKDPDVLITYNGDNFDFAYLKKRCEELGIKFTLGRDGSEPKIQRMGDRFAVEVKGRIHFDLYPVIRRTINLPTYTLEAVYEAVFGKPKEKVYAEEIAQAWESGEGLERVARYSMEDAKVTYELGREFFPMEAQLSRLIGQSLWDVSRSSTGNLVEWFLLRKAYKRNELAPNKPDERELARRRGGYAGGYVKEPERGLWDNIVYLDFRSLYPSIIITHNVSPDTLNREGCKEYDVAPEVGHKFCKDFPGFIPSLLGDLLEERQKIKRKMKATVDPLEKKLLDYRQRAIKILANSFYGYYGYAKARWYCKECAESVTAWGREYIEMVIRELEEKFGFKVLYADTDGLHATIPGADAETVKKKAKEFLKYINPKLPGLLELEYEGFYVRGFFVTKKKYAVIDEEGKITTRGLEIVRRDWSEIAKETQARVLEAILKHGDVEEAVRIVKEVTEKLSKYEVPPEKLVIHEQITRDLRDYKATGPHVAVAKRLAARGVKIRPGTVISYIVLKGSGRIGDRAIPADEFDPTKHRYDAEYYIENQVLPAVERILKAFGYRKEDLRYQKTKQVGLGAWLKVKGKK* +ECOpolA: + description: DNA polymerase that also exhibits 3'-5' and 5'-3' exonuclease activity. + sequence: MVQIPQNPLILVDGSSYLYRAYHAFPPLTNSAGEPTGAMYGVLNMLRSLIMQYKPTHAAVVFDAKGKTFRDELFEHYKSHRPPMPDDLRAQIEPLHAMVKAMGLPLLAVSGVEADDVIGTLAREAEKAGRPVLISTGDKDMAQLVTPNITLINTMTNTILGPEEVVNKYGVPPELIIDFLALMGDSSDNIPGVPGVGEKTAQALLQGLGGLDTLYAEPEKIAGLSFRGAKTMAAKLEQNKEVAYLSYQLATIKTDVELELTCEQLEVQQPAAEELLGLFKKYEFKRWTADVEAGKWLQAKGAKPAAKPQETSVADEAPEVTATVISYDNYVTILDEETLKAWIAKLEKAPVFAFDTETDSLDNISANLVGLSFAIEPGVAAYIPVAHDYLDAPDQISRERALELLKPLLEDEKALKVGQNLKYDRGILANYGIELRGIAFDTMLESYILNSVAGRHDMDSLAERWLKHKTITFEEIAGKGKNQLTFNQIALEEAGRYAAEDADVTLQLHLKMWPDLQKHKGPLNVFENIEMPLVPVLSRIERNGVKIDPKVLHNHSEELTLRLAELEKKAHEIAGEEFNLSSTKQLQTILFEKQGIKPLKKTPGGAPSTSEEVLEELALDYPLPKVILEYRGLAKLKSTYTDKLPLMINPKTGRVHTSYHQAVTATGRLSSTDPNLQNIPVRNEEGRRIRQAFIAPEDYVIVSADYSQIELRIMAHLSRDKGLLTAFAEGKDIHRATAAEVFGLPLETVTSEQRRSAKAINFGLIYGMSAFGLARQLNIPRKEAQKYMDLYFERYPGVLEYMERTRAQAKEQGYVETLDGRRLYLPDIKSSNGARRAAAERAAINAPMQGTAADIIKRAMIAVDAWLQAEQPRVRMIMQVHDELVFEVHKDDVDAVAKQIHQLMENCTRLDVPLLVEVGSGENWDQAH* +KTHEAQpol: + description: Exonuclease-free, N-terminal deletion mutant of Taq DNA polymerase. + sequence: MGLLHEFGLLESPKALEEAPWPPPEGAFVGFVLSRKEPMWADLLALAAARGGRVHRAPEPYKALRDLKEARGLLAKDLSVLALREGLGLPPGDDPMLLAYLLDPSNTTPEGVARRYGGEWTEEAGERAALSERLFANLWGRLEGEERLLWLYREVERPLSAVLAHMEATGVRLDVAYLRALSLEVAEEIARLEAEVFRLAGHPFNLNSRDQLERVLFDELGLPAIGKTEKTGKRSTSAAVLEALREAHPIVEKILQYRELTKLKSTYIDPLPDLIHPRTGRLHTRFNQTATATGRLSSSDPNLQNIPVRTPLGQRIRRAFIAEEGWLLVALDYSQIELRVLAHLSGDENLIRVFQEGRDIHTETASWMFGVPREAVDPLMRRAAKTINFGVLYGMSAHRLSQELAIPYEEAQAFIERYFQSFPKVRAWIEKTLEEGRRRGYVETLFGRRRYVPDLEARVKSVREAAERMAFNMPVQGTAADLMKLAMVKLFPRLEEMGARMLLQVHDELVLEAPKERAEAVARLAKEVMEGVYPLAVPLEVEVGIGEDWLSAKE* +9N7polA_CT: + description: 9 Degrees North DNA polymerase variant with enhanced ability to incorporate modified substrates + sequence: MILDTDYITENGKPVIRVFKKENGEFKIEYDRTFEPYFYALLKDDSAIEDVKKVTAKRHGTVVKVKRAEKVQKKFLGRPIEVWKLYFNHPQDVPAIRDRIRAHPAVVDIYEYDIPFAKRYLIDKGLIPMEGDEELTMLAFAIATLYHEGEEFGTGPILMISYADGSEARVITWKKIDLPYVDVVSTEKEMIKRFLRVVREKDPDVLITYNGDNFDFAYLKKRCEELGIKFTLGRDGSEPKIQRMGDRFAVEVKGRIHFDLYPVIRRTINLPTYTLEAVYEAVFGKPKEKVYAEEIAQAWESGEGLERVARYSMEDAKVTYELGREFFPMEAQLSRLIGQSLWDVSRSSTGNLVEWFLLRKAYKRNELAPNKPDERELARRRGGYAGGYVKEPERGLWDNIVYLDFRSSAVSIIITHNVSPDTLNREGCKEYDVAPEVGHKFCKDFPGFIPSLLGDLLEERQKIKRKMKATVDPLEKKLLDYRQRAIKILANSFYGYYGYAKARWYCKECAESVTAWGREYIEMVIRELEEKFGFKVLYADTDGLHATIPGADAETVKKKAKEFLKYINPKLPGLLELEYEGFYVRGFFVTKKKYAVIDEEGKITTRGLEIVRRDWSEIAKETQARVLEAILKHGDVEEAVRIVKEVTEKLSKYEVPPEKLVIHEQITRDLRDYKATGPHVAVAKRLAARGVKIRPGTVISYIVLKGSGRIGDRAIPADEFDPTKHRYDAEYYIENQVLPAVERILKAFGYRKEDLRYQKTKQVGLGAWLKVKGKK* +dbh: + description: Thermostable gamma-family lesion-bypass DNA Polymerase that efficiently synthesizes DNA across a variety of DNA template lesions. + sequence: MIVLFVDFDYFYAQVEEVLNPSLKGKPVVVCVFSGRFEDSGAVATANYEARKFGVKAGIPIVEAKKILPNAVYLPMRKEVYQQVSSRIMNLLREYSEKIEIASIDEAYLDISDKVRDYREAYNLGLEIKNKILEKEKITVTVGISKNKVFAKIAADMAKPNGIKVIDDEEVKRLIRELDIADVPGIGNITAEKLKKLGINKLVDTLSIEFDKLKGMIGEAKAKYLISLARDEYNEPIRTRVRKSIGRIVTMKRNSRNLEEIKPYLFRAIEESYYKLDKRIPKAIHVVAVTEDLDIVSRGRTFPHGISKETAYSESVKLLQKILEEDERKIRRIGVRFSKFIEAIGLDKFFDT* +T7gene5: + description: DNA Pol with high fidelity and rapid extension rate (useful in copying long stretches of DNA); Has strong 3'-5' exonuclease. + sequence: MIVSDIEANALLESVTKFHCGVIYDYSTAEYVSYRPSDFGAYLDALEAEVARGGLIVFHNGHKYDVPALTKLAKLQLNREFHLPRENCIDTLVLSRLIHSNLKDTDMGLLRSGKLPGKRFGSHALEAWGYRLGEMKGEYKDDFKRMLEEQGEEYVDGMEWWNFNEEMMDYNVQDVVVTKALLEKLLSDKHYFPPEIDFTDVGYTTFWSESLEAVDIEHRAAWLLAKQERNGFPFDTKAIEELYVELAARRSELLRKLTETFGSWYQPKGGTEMFCHPRTGKPLPKYPRIKTPKVGGIFKKPKNKAQREGREPCELDTREYVAGAPYTPVEHVVFNPSSRDHIQKKLQEAGWVPTKYTDKGAPVVDDEVLEGVRVDDPEKQAAIDLIKEYLMIQKRIGQSAEGDKAWLRYVAEDGKIHGSVNPNGAVTGRATHAFPNLAQIPGVRSPYGEQCRAAFGAEHHLDGITGKPWVQAGIDASGLELRCLAHFMARFDNGEYAHEILNGDIHTKNQIAAELPTRDNAKTFIYGFLYGAGDEKIGQIVGAGKERGKELKKKFLENTPAIAALRESIQQTLVESSQWVAGEQQVKWKRRWIKGLDGRKVHVRSPHAALNTLLQSAGALICKLWIIKTEEMLVEKGLKHGWDGDFAYMAWVHDEIQVGCRTEEIAQVVIETAQEAMRWVGDHWNFRCLLDTEGKMGPNWAICH* # Reverse Transcriptase +TthRT: + description: DNA polymerase that efficiently reverse-transcribes RNA in the presence of MnCl2 + sequence: MEAMLPLFEPKGRVLLVDGHHLAYRTFFALKGLTTSRGEPVQAVYGFAKSLLKALKEDGYKAVFVVFDAKAPSFRHEAYEAYKAGRAPTPEDFPRQLALIKELVDLLGFTRLEVPGYEADDVLATLAKKAEKEGYEVRILTADRDLYQLVSDRVAVLHPEGHLITPEWLWEKYGLRPEQWVDFRALVGDPSDNLPGVKGIGEKTALKLLKEWGSLENLLKNLDRVKPENVREKIKAHLEDLRLSLELSRVRTDLPLEVDLAQGREPDREGLRAFLERLEFGSLLHEFGLLEAPAPLEEAPWPPPEGAFVGFVLSRPEPMWAELKALAACRDGRVHRAADPLAGLKDLKEVRGLLAKDLAVLASREGLDLVPGDDPMLLAYLLDPSNTTPEGVARRYGGEWTEDAAHRALLSERLHRNLLKRLEGEEKLLWLYHEVEKPLSRVLAHMEATGVRRDVAYLQALSLELAEEIRRLEEEVFRLAGHPFNLNSRDQLERVLFDELRLPALGKTQKTGKRSTSAAVLEALREAHPIVEKILQHRELTKLKNTYVDPLPSLVHPRTGRLHTRFNQTATATGRLSSSDPNLQNIPVRTPLGQRIRRAFVAEAGWALVALDYSQIELRVLAHLSGDENLIRVFQEGKDIHTQTASWMFGVPPEAVDPLMRRAAKTVNFGVLYGMSAHRLSQELAIPYEEAVAFIERYFQSFPKVRAWIEKTLEEGRKRGYVETLFGRRRYVPDLNARVKSVREAAERMAFNMPVQGTAADLMKLAMVKLFPRLREMGARMLLQVHDELLLEAPQARAEEVAALAKEAMEKAYPLAVPLEVEVGMGEDWLSAKG* +MMLV_RT_mut_H: + description: Single 75 kDa monomer, cDNA synthesis; high enzyme activity and processivity. + sequence: MTLNIEDEHRLHETSKEPDVSLGSTWLSDFPQAWAETGGMGLAVRQAPLIIPLKATSTPVSIKQYPMSQEARLGIKPHIQRLLDQGILVPCQSPWNTPLLPVKKPGTNDYRPVQDLREVNKRVEDIHPTVPNPYNLLSGLPPSHQWYTVLDLKDAFFCLRLHPTSQPLFAFEWRDPEMGISGQLTWTRLPQGFKNSPTLFDEALHRDLADFRIQHPDLILLQYVDDLLLAATSELDCQQGTRALLQTLGNLGYRASAKKAQICQKQVKYLGYLLKEGQRWLTEARKETVMGQPTPKTPRQLREFLGTAGFCRLWIPGFAEMAAPLYPLTKTGTLFNWGPDQQKAYQEIKQALLTAPALGLPDLTKPFELFVDEKQGYAKGVLTQKLGPWRRPVAYLSKKLDPVAAGWPPCLRMVAAIAVLTKDAGKLTMGQPLVILAPHAVEALVKQPPDRWLSNARMTHYQALLLDTDRVQFGPVVALNPATLLPLPEEGLQHNCLDILAEAHGTRPDLTDQPLPDADHTWYTGGSSLLQEGQRKAGAAVTTETEVIWAKALPAGTSAQRAQLIALTQALKMAEGKKLNVYTNSRYAFATAHIHGEIYRRRGLLTSEGKEIKNKDEILALLKALFLPKRLSIIHCPGHQKGHSAEARGNRMADQAARKAA* +MMLV_RT_lack_H: + description: Single 75 kDa monomer, cDNA synthesis; high enzyme activity and processivity. Comparable to SuperScriptII from Thermo Fisher. + sequence: MTLNIEDEHRLHETSKEPDVSLGSTWLSDFPQAWAETGGMGLAVRQAPLIIPLKATSTPVSIKQYPMSQEARLGIKPHIQRLLDQGILVPCQSPWNTPLLPVKKPGTNDYRPVQDLREVNKRVEDIHPTVPNPYNLLSGLPPSHQWYTVLDLKDAFFCLRLHPTSQPLFAFEWRDPEMGISGQLTWTRLPQGFKNSPTLFDEALHRDLADFRIQHPDLILLQYVDDLLLAATSELDCQQGTRALLQTLGNLGYRASAKKAQICQKQVKYLGYLLKEGQRWLTEARKETVMGQPTPKTPRQLREFLGTAGFCRLWIPGFAEMAAPLYPLTKTGTLFNWGPDQQKAYQEIKQALLTAPALGLPDLTKPFELFVDEKQGYAKGVLTQKLGPWRRPVAYLSKKLDPVAAGWPPCLRMVAAIAVLTKDAGKLTMGQPLVILAPHAVEALVKQPPDRWLSNARMTHYQALLLDTDRVQFGPVVALNPATLLPLPEEGLQHNCLDNSRLI* # RNA polymerase +T3_RNAP: + description: DNA-dependent RNA polymerase derived from T3 bacteriophage that catalyzes the transcription of DNA into RNA using the four ribonucleoside triphosphates as substrates. + sequence: MNIIENIEKNDFSEIELAAIPFNTLADHYGSALAKEQLALEHESYELGERRFLKMLERQAKAGEIADNAAAKPLLATLLPKLTTRIVEWLEEYASKKGRKPSAYAPLQLLKPEASAFITLKVILASLTSTNMTTIQAAAGMLGKAIEDEARFGRIRDLEAKHFKKHVEEQLNKRHGQVYKKAFMQVVEADMIGRGLLGGEAWSSWDKETTMHVGIRLIEMLIESTGLVELQRHNAGNAGSDHEALQLAQEYVDVLAKRAGALAGISPMFQPCVVPPKPWVAITGGGYWANGRRPLALVRTHSKKGLMRYEDVYMPEVYKAVNLAQNTAWKINKKVLAVVNEIVNWKNCPVADIPSLERQELPPKPDDIDTNEAALKEWKKAAAGIYRLDKARVSRRISLEFMLEQANKFASKKAIWFPYNMDWRGRVYAVPMFNPQGNDMTKGLLTLAKGKPIGEEGFYWLKIHGANCAGVDKVPFPERIAFIEKHVDDILACAKDPINNTWWAEQDSPFCFLAFCFEYAGVTHHGLSYNCSLPLAFDGSCSGIQHFSAMLRDEVGGRAVNLLPSETVQDIYGIVAQKVNEILKQDAINGTPNEMITVTDKDTGEISEKLKLGTSTLAQQWLAYGVTRSVTKRSVMTLAYGSKEFGFRQQVLDDTIQPAIDSGKGLMFTQPNQAAGYMAKLIWDAVSVTVVAAVEAMNWLKSAAKLLAAEVKDKKTKEILRHRCAVHWTTPDGFPVWQEYRKPLQKRLDMIFLGQFRLQPTINTLKDSGIDAHKQESGIAPNFVHSQDGSHLRMTVVYAHEKYGIESFALIHDSFGTIPADAGKLFKAVRETMVITYENNDVLADFYSQFADQLHETQLDKMPPLPKKGNLNLQDILKSDFAFA* +T7_RNAP: + description: Highly processive DNA-dependent RNA polymerase derived from T7 bacteriophage + sequence: MNTINIAKNDFSDIELAAIPFNTLADHYGERLAREQLALEHESYEMGEARFRKMFERQLKAGEVADNAAAKPLITTLLPKMIARINDWFEEVKAKRGKRPTAFQFLQEIKPEAVAYITIKTTLACLTSADNTTVQAVASAIGRAIEDEARFGRIRDLEAKHFKKNVEEQLNKRVGHVYKKAFMQVVEADMLSKGLLGGEAWSSWHKEDSIHVGVRCIEMLIESTGMVSLHRQNAGVVGQDSETIELAPEYAEAIATRAGALAGISPMFQPCVVPPKPWTGITGGGYWANGRRPLALVRTHSKKALMRYEDVYMPEVYKAINIAQNTAWKINKKVLAVANVITKWKHCPVEDIPAIEREELPMKPEDIDMNPEALTAWKRAAAAVYRKDKARKSRRISLEFMLEQANKFANHKAIWFPYNMDWRGRVYAVSMFNPQGNDMTKGLLTLAKGKPIGKEGYYWLKIHGANCAGVDKVPFPERIKFIEENHENIMACAKSPLENTWWAEQDSPFCFLAFCFEYAGVQHHGLSYNCSLPLAFDGSCSGIQHFSAMLRDEVGGRAVNLLPSETVQDIYGIVAKKVNEILQADAINGTDNEVVTVTDENTGEISEKVKLGTKALAGQWLAYGVTRSVTKRSVMTLAYGSKEFGFRQQVLEDTIQPAIDSGKGLMFTQPNQAAGYMAKLIWESVSVTVVAAVEAMNWLKSAAKLLAAEVKDKKTGEILRKRCAVHWVTPDGFPVWQEYKKPIQTRLNLMFLGQFRLQPTINTNKDSEIDAHKQESGIAPNFVHSQDGSHLRKTVVWAHEKYGIESFALIHDSFGTIPADAANLFKAVRETMVDTYESCDVLADFYDQFADQLHESQLDKMPALPAKGNLNLRDILESDFAFA* +SP6_RNAP: + description: DNA-dependent RNA polymerase derived from SP6 bacteriophage that catalyzes the transcription of viral genes. + sequence: MQDLHAIQLQLEEEMFNGGIRRFEADQQRQIAAGSESDTAWNRRLLSELIAPMAEGIQAYKEEYEGKKGRAPRALAFLQCVENEVAAYITMKVVMDMLNTDATLQAIAMSVAERIEDQVRFSKLEGHAAKYFEKVKKSLKASRTKSYRHAHNVAVVAEKSVAEKDADFDRWEAWPKETQLQIGTTLLEILEGSVFYNGEPVFMRAMRTYGGKTIYYLQTSESVGQWISAFKEHVAQLSPAYAPCVIPPRPWRTPFNGGFHTEKVASRIRLVKGNREHVRKLTQKQMPKVYKAINALQNTQWQINKDVLAVIEEVIRLDLGYGVPSFKPLIDKENKPANPVPVEFQHLRGRELKEMLSPEQWQQFINWKGECARLYTAETKRGSKSAAVVRMVGQARKYSAFESIYFVYAMDSRSRVYVQSSTLSPQSNDLGKALLRFTEGRPVNGVEALKWFCINGANLWGWDKKTFDVRVSNVLDEEFQDMCRDIAADPLTFTQWAKADAPYEFLAWCFEYAQYLDLVDEGRADEFRTHLPVHQDGSCSGIQHYSAMLRDEVGAKAVNLKPSDAPQDIYGAVAQVVIKKNALYMDADDATTFTSGSVTLSGTELRAMASAWDSIGITRSLTKKPVMTLPYGSTRLTCRESVIDYIVDLEEKEAQKAVAEGRTANKVHPFEDDRQDYLTPGAAYNYMTALIWPSISEVVKAPIVAMKMIRQLARFAAKRNEGLMYTLPTGFILEQKIMATEMLRVRTCLMGDIKMSLQVETDIVDEAAMMGAAAPNFVHGHDASHLILTVCELVDKGVTSIAVIHDSFGTHADNTLTLRVALKGQMVAMYIDGNALQKLLEEHEVRWMVDTGIEVPEQGEFDLNEIMDSEYVFA* # Other +BovDNTT: + description: Adds dT at the end of PCR blunted DNA fragments/vectors that can then be used in TOPO cloning; Addition of homopolymer tails to the 3' ends of DNA; Labeling the 3' ends of DNA with modified nucleotides (e.g., ddNTP, DIG-dUTP); TUNEL assay (in situ localization of apoptosis); TdT dependent PCR + sequence: MDPLCTASSGPRKKRPRQVGASMASPPHDIKFQNLVLFILEKKMGTTRRNFLMELARRKGFRVENELSDSVTHIVAENNSGSEVLEWLQVQNIRASSQLELLDVSWLIESMGAGKPVEITGKHQLVVRTDYSATPNPGFQKTPPLAVKKISQYACQRKTTLNNYNHIFTDAFEILAENSEFKENEVSYVTFMRAASVLKSLPFTIISMKDTEGIPCLGDKVKCIIEEIIEDGESSEVKAVLNDERYQSFKLFTSVFGVGLKTSEKWFRMGFRSLSKIMSDKTLKFTKMQKAGFLYYEDLVSCVTRAEAEAVGVLVKEAVWAFLPDAFVTMTGGFRRGKKIGHDVDFLITSPGSAEDEEQLLPKVINLWEKKGLLLYYDLVESTFEKFKLPSRQVDTLDHFQKCFLILKLHHQRVDSSKSNQQEGKTWKAIRVDLVMCPYENRAFALLGWTGSRQFERDIRRYATHERKMMLDNHALYDKTKRVFLKAESEEEIFAHLGLDYIEPWERNA* +T4-BGT: + description: T4 Phage beta-glucosyltransferasetransfers the glucose moiety of UDP-Glc to the 5-hmC residues in double-stranded DNA. + sequence: MKIAIINMGNNVINFKTVPSSETIYLFKVISEMGLNVDIISLKNGVYTKSFDEVDVNDYDRLIVVNSSINFFGGKPNLAILSAQKFMAKYKSKIYYLFTDIRLPFSQSWPNVKNRPWAYLYTEEELLIKSPIKVISQGINLDIAKAAHKKVDNVIEFEYFPIEQYKIHMNDFQLSKPTKKTLDVIYGGSFRSGQRESKMVEFLFDTGLNIEFFGNAREKQFKNPKYPWTKAPVFTGKIPMNMVSEKNSQAIAALIIGDKNYNDNFITLRVWETMASDAVMLIDEEFDTKHRIINDARFYVNNRAELIDRVNELKHSDVLRKEMLSIQHDILNKTRAKKAEWQDAFKKAIDL* +Endonuclease_VIII_Ecoli: + description: Bifunctional DNA glycosylase with DNA N-glycosylase and AP lyase activities; The N-glycosylase activity releases damaged pyrimidines, including thymine glycol and uracil glycol. The AP lyase activity cleaves DNA phosphodiester backbone at AP sites via beta and delta-elimination, creating a 1 nucleotide DNA gap with 5' and 3' phosphate termini. + sequence: MPEGPEIRRAADNLEAAIKGKPLTDVWFAFPQLKPYQSQLIGQHVTHVETRGKALLTHFSNDLTLYSHNQLYGVWRVVDTGEEPQTTRVLRVKLQTADKTILLYSASDIEMLTPEQLTTHPFLQRVGPDVLDPNLTPEVVKERLLSPRFRNRQFAGLLLDQAFLAGLGNYLRVEILWQVGLTGNHKAKDLNAAQLDALAHALLEIPRFSYATRGQVDENKHHGALFRFKVFHRDGEPCERCGSIIEKTTLSSRPFYWCPGCQH* +dromedNK: + description: Used for dNTP synthesis + sequence: MAEAASCARKGTKYAEGTQPFTVLIEGNIGSGKTTYLNHFEKYKNDICLLTEPVEKWRNVNGVNLLELMYKDPKKWAMPFQSYVTLTMLQSHTAPTNKKLKIMERSIFSARYCFVENMRRNGSLEQGMYNTLEEWYKFIEESIHVQADLIIYLRTSPEVAYERIRQRARSEESCVPLKYLQELHELHEDWLIHQRRPQSCKVLVLDADLNLENIGTEYQRSESSIFDAISSNQQPSPVLVSPSKRQRVAR* +DNA_Topoisomerase_1B_Vaccinia_Virus: + description: Used in TOPO cloning. Recognizes the DNA sequence 5'-(C/T)CCTT-3' and digests double stranded DNA at this sequence. + sequence: MRALFYKDGKLFTDNNFLNPVSDDNPAYEVLQHVKIPTHLTDVVVYEQTWEEALTRLIFVGSDSKGRRQYFYGKMHVQNRNAKRDRIFVRVYNVMKRINCFINKNIKKSSTDSNYQLAVFMLMETMFFIRFGKMKYLKENETVGLLTLKNKHIEISPDEIVIKFVGKDKVSHEFVVHKSNRLYKPLLKLTDDSSPEEFLFNKLSERKVYECIKQFGIRIKDLRTYGVNYTFLYNFWTNVKSISPLPSPKKLIALTIKQTAEVVGHTPSISKRAYMATTILEMVKDKNFLDVVSKTTFDEFLSIVVDHVKSSTDG* +T7EI: + description: DNA Endonuclease; catalyzes cleavage of DNA mismatches and non-beta DNA structures (Holliday junctions and cruciform). + sequence: MAGYGAKGIRKVGAFRSGLEDKVSKQLESKGIKFEYEEWKVPYVIPASNHTYTPDFLLPNGIFVETKGLWESDDRKKHLIIREQHPELDIRIVFSSSRTKLYKGSPTSYGEFCEKHGIKFADKLIPAEWIKEPKKEVPFDRLKRKGGKK* +TEV_protease: + description: Used for site-specific cleavage of proteins. TEV cleaves the protein consensus sequence EXXYXQ^S/G + sequence: GESLFKGPRDYNPISSTICHLTNESDGHTTSLYGIGFGPFIITNKHLFRRNNGTLLVQSLHGVFKVKNTTTLQQHLIDGRDMIIIRMPKDFPPFPQKLKFREPQREERICLVTTNFQTKSMSSMVSDTSCTFPSSDGIFWKHWIQTKDGQCGSPLVSTRDGFIVGIHSASNFTNTNNYFTSVPKNFMELLTNQEAQQWVSGWRLNADSVLWGGHKVFMVKPEEPFQPVKEATQLMN +RNAse_H: + description: Endonuclease that specifically degrades the RNA of RNA-DNA hybrids. Participates in DNA replication; helps to specify the origin of genomic replication. + sequence: MLKQVEIFTDGSCLGNPGPGGYGAILRYRGREKTFSAGYTRTTNNRMELMAAIVALEALKEHCEVILSTDSQYVRQGITQWIHNWKKRGWKTADKKPVKNVDLWQRLDAALGQHQIKWEWVKGHAGHPENERCDELARAAAMNPTLEDTGYQVEV* +ALPI: + description: CIAP reporter; can be used in spectrophotometric or luminescence assay when appropriate substrate is used + sequence: MQGACVLLLLGLHLQLSLGLVPVEEEDPAFWNRQAAQALDVAKKLQPIQTAAKNVILFLGDGMGVPTVTATRILKGQMNGKLGPETPLAMDQFPYVALSKTYNVDRQVPDSAGTATAYLCGVKGNYRTIGVSAAARYNQCKTTRGNEVTSVMNRAKKAGKSVGVVTTTRVQHASPAGAYAHTVNRNWYSDADLPADAQMNGCQDIAAQLVNNMDIDVILGGGRKYMFPVGTPDPEYPDDASVNGVRKRKQNLVQAWQAKHQGAQYVWNRTALLQAADDSSVTHLMGLFEPADMKYNVQQDHTKDPTLQEMTEVALRVVSRNPRGFYLFVEGGRIDHGHHDDKAYMALTEAGMFDNAIAKANELTSELDTLILVTADHSHVFSFGGYTLRGTSIFGLAPSKALDSKSYTSILYGNGPGYALGGGSRPDVNDSTSEDPSYQQQAAVPQASETHGGEDVAVFARGPQAHLVHGVEEETFVAHIMAFAGCVEPYTDCNLPAPTTATSIPDAAHLAASPPPLALLAGAMLLLLAPTLY* +RNAse_A: + description: Ribonuclease A or RNase A; endoribonuclease purified from bovine pancreas. Important enzyme for the removal of RNA for RNA free DNA purification reactions such as plasmid DNA purification and genomic DNA purification, RNA removal from recombinant protein preparations, ribonuclease protection assays, mapping single-base mutations in DNA/RNA. + sequence: MALKSLVLLSLLVLVLLLVRVQPSLGKETAAAKFERQHMDSSTSAASSSNYCNQMMKSRNLTKDRCKPVNTFVHESLADVQAVCSQKNVACKNGQTNCYQSYSTMSITDCRETGSSKYPNCAYKTTQANKHIIVACEGNPYVPVHFDASV* +T5gene12_13: + description: Double-stranded DNA specific exonuclease and single-stranded DNA endonuclease. Initiates at the 5' termini of linear or nicked double-stranded DNA. Cleaves linear or nicked double-stranded DNA in the 5' to 3' direction. + sequence: MSKSWGKFIEEEEAEMASRRNLMIVDGTNLGFRFKHNNSKKPFASSYVSTIQSLAKSYSARTTIVLGDKGKSVFRLEHLPEYKGNRDEKYAQRTEEEKALDEQFFEYLKDAFELCKTTFPTFTIRGVEADDMAAYIVKLIGHLYDHVWLISTDGDWDTLLTDKVSRFSFTTRREYHLRDMYEHHNVDDVEQFISLKAIMGDLGDNIRGVEGIGAKRGYNIIREFGNVLDIIDQLPLPGKQKYIQNLNASEELLFRNLILVDLPTYCVDAIAAVGQDVLDKFTKDILEIAEQ* +T4_PGD: + description: T4 pyrimidine DNA glycosylase; bifunctional DNA glycosylase with DNA N-glycosylase and AP lyase activities. + sequence: MTRINLTLVSELADQHLMAEYRELPRVFGAVRKHVANGKRVRDFKISPTFILGAGHVTFFYDKLEFLRKRQIELIAECLKRGFNIKDTTVQDISDIPQEFRGDYIPHEASIAISQARLDEKIAQRPTWYKYYGKAIYA* +T4_g32: + description: A single-stranded DNA binding protein required for bacteriophage T4 replication and repair; Used extensively to stabilize and mark regions of ssDNA for electron microscopic examination of intracellular DNA structure, to increase yield and processivity of reverse transcriptase during RT-PCR, and to increase yield and specificity of PCR products from soil samples. + sequence: MFKRKSTAELAAQMAKLNGNKGFSSEDKGEWKLKLDNAGNGQAVIRFLPSKNDEQAPFAILVNHGFKKNGKWYIETCSSTHGDYDSCPVCQYISKNDLYNTDNKEYSLVKRKTSYWANILVVKDPAAPENEGKVFKYRFGKKIWDKINAMIAVDVEMGETPVDVTCPWEGANFVLKVKQVSGFSNYDESKFLNQSAIPNIDDESFQKELFEQMVDLSEMTSKDKFKSFEELNTKFGQVMGTAVMGGAAATAAKKADKVADDLDAFNVDDFNTKTEDDFMSSSSGSSSSADDTDLDDLLNDL* +Exonuclease_III_Ecoli: + description: Double-stranded DNA specific exonuclease. Initiates at the 3' termini of linear double-stranded DNA with 5' overhangs or blunt ends and 3' overhangs containing less than four bases. Initiates at nicked sites in double-stranded DNA. + sequence: MKFVSFNINGLRARPHQLEAIVEKHQPDVIGLQETKVHDDMFPLEEVAKLGYNVFYHGQKGHYGVALLTKETPIAVRRGFPGDDEEAQRRIIMAEIPSLLGNVTVINGYFPQGESRDHPIKFPAKAQFYQNLQNYLETELKRDNPVLIMGDMNISPTDLDIGIGEENRKRWLRTGKCSFLPEEREWMDRLMSWGLVDTFRHANPQTADRFSWFDYRSKGFDDNRGLRIDLLLASQPLAECCVETGIDYEIRSMEKPSDHAPVWATFRR* +UDG: + description: Uracil DNA glycosylase catalyzes the hydrolysis of the N-glycosidic bond from deoxyuridine to release uracil. + sequence: MANELTWHDVLAEEKQQPYFLNTLQTVASERQSGVTIYPPQKDVFNAFRFTELGDVKVVILGQDPYHGPGQAHGLAFSVRPGIAIPPSLLNMYKELENTIPGFTRPNHGYLESWARQGVLLLNTVLTVRAGQAHSHASLGWETFTDKVISLINQHREGVVFLLWGSHAQKKGAIIDKQRHHVLKAPHPSPLSAHRGFFGCNHFVLANQWLEQRGETPIDWMPVLPAESE* +ETSSB: + description: ssDNA binding protein, 9kDa. Features - Improves the processivity of DNA polymerase; involved in stabilization and marking of ssDNA structure; increases the yield and specificitiy of PCR; increases the yield and processivity of RT during RT-PCR; improves DNA sequencing through regions with strong secondary structure + sequence: MEEKVGNLKPNMESVNVTVRVLEASEARQIQTKNGVRTISEAIVGDETGRVKLTLWGKHAGSIKEGQVVKIENAWTTAFKGQVQLNAGSKTKIAEASEDGFPESSQIPENTPTAPQQMRGGGRGFRGGGRRYGRRGGRRQENEEGEEE* +Fpg: + description: Formamidopyrimidine DNA Glycosylase; Bifunctional DNA glycosylase with DNA N-glycosylase and AP lyase activities + sequence: MPELPEVETSRRGIEPHLVGATILHAVVRNGRLRWPVSEEIYRLSDQPVLSVQRRAKYLLLELPEGWIIIHLGMSGSLRILPEELPPEKHDHVDLVMSNGKVLRYTDPRRFGAWLWTKELEGHNVLTHLGPEPLSDDFNGEYLHQKCAKKKTAIKPWLMDNKLVVGVGNIYASESLFAAGIHPDRLASSLSLAECELLARVIKAVLLRSIEQGGTTLKDFLQSDGKPGYFAQELQVYGRKGEPCRVCGTPIVATKHAQRATFYCRQCQK* +Endonuclease_IV_E_coli: + description: DNA AP endonuclease; Catalyzes the cleavage of DNA phosphodiester backbone at AP sites via hydrolysis leaving a 1 nucleotide gap with 3'-hydroxyl and 5' deoxyribose phosphate (dRP) termini; Also has 3'-diesterase activity which can remove 3' phosphate, 3'-alpha, beta-unsaturated aldehyde, phosphoglycoaldehyde, and other 3' blocking groups. + sequence: MKYIGAHVSAAGGLANAAIRAAEIDATAFALFTKNQRQWRAAPLTTQTIDEFKAACEKYHYTSAQILPHDSYLINLGHPVTEALEKSRDAFIDEMQRCEQLGLSLLNFHPGSHLMQISEEDCLARIAESINIALDKTQGVTAVIENTAGQGSNLGFKFEHLAAIIDGVEDKSRVGVCIDTCHAFAAGYDLRTPAECEKTFADFARTVGFKYLRGMHLNDAKSTFGSRVDRHHSLGEGNIGHDAFRWIMQDDRFDGIPLILETINPDIWAEEIAWLKAQQTEKAVA* +T4_PNK: + description: The most commonly used polynucleotide kinase. + sequence: MKKIILTIGCPGSGKSTWAREFIAKNPGFYNINRDDYRQSIMAHEERDEYKYTKKKEGIVTGMQFDTAKSILYGGDSVKGVIISDTNLNPERRLAWETFAKEYGWKVEHKVFDVPWTELVKRNSKRGTKAVPIDVLRSMYKSMREYLGLPVYNGTPGKPKAVIFDVDGTLAKMNGRGPYDLEKCDTDVINPMVVELSKMYALMGYQIVVVSGRESGTKEDPTKYYRMTRKWVEDIAGVPLVMQCQREQGDTRKDDVVKEEIFWKHIAPHFDVKLAIDDRTQVVEMWRRIGVECWQVASGDF* From 3592666097db35831d14cd9107b0555fc31d395d Mon Sep 17 00:00:00 2001 From: Keoni Gandall Date: Tue, 29 Oct 2024 09:51:36 -0700 Subject: [PATCH 9/9] in-between --- parts/linkers2.md | 110 ++++++++++++++++++++++++++++++++++++++++++++-- 1 file changed, 106 insertions(+), 4 deletions(-) diff --git a/parts/linkers2.md b/parts/linkers2.md index f1f3e4b..c1e50dd 100644 --- a/parts/linkers2.md +++ b/parts/linkers2.md @@ -1,16 +1,23 @@ # DnaDesign Assembly -DnaDesign Assembly (shortened as dd assembly) is a GoldenGate DNA assembly method similar to MoClo. The overhang set was changed from MoClo in order to accomodate the overhangs `GTCT` and `CGAG`, which are used in recursive DNA assemblies. The BsaI overhangs were optimized using [empirical data](https://doi.org/10.1371/journal.pone.0238592) from the paper "Enabling one-pot Golden Gate assemblies of unprecedented complexity using data-optimized assembly design". +DnaDesign Assembly (shortened as dd assembly) is a GoldenGate DNA assembly method similar to MoClo. It is designed from the bottom up for manufacturing with the intention of globally lowering the cost of useful synthetic DNA. The overhang set was changed from MoClo in order to accomodate the overhangs `GTCT` and `CGAG`, which are used in recursive DNA assemblies. The BsaI overhangs were optimized using [empirical data](https://doi.org/10.1371/journal.pone.0238592) from the paper "Enabling one-pot Golden Gate assemblies of unprecedented complexity using data-optimized assembly design". + +## Simplest user perspective + +From the simplest user perspective, dd assembly has genetic parts. These parts can be put together in a modular fashion into much larger genetic designs. Synthesis and assembly time is minimized, while allowing the user to do almost anything they want - from creating fusion proteins, transcriptional units, operons, and shuttle vectors. If they do not require novel synthesis, the goal of dd assembly is to allow time from ordering to recieving clonal DNA within 3 days (3 day plasmids), regardless of the size or complexity. + +## Overhangs overhangs: `CGAG,GTCT,GGGG,AAAA,AACT,AATG,ATCC,CGCT,TTCT,AAGC,ATAG,ATTA,ATGT,ACTC,ACGA,TATC,TAGG,TACA,TTAC,TTGA,TGGA,GAAG,GACC,GCCG` overhangs with numbers: -1. CGAG +X. GTCT +Y. CGAG 2. TACA 3. AACT 4. AATG 5. ATCC 6. CGCT -7. GTCT +7. TACA 8. AAGC 9. ATAG 10. ATTA @@ -30,6 +37,51 @@ overhangs with numbers: 24. AAAA 25. GGGG +This document is techincal reference material, not a how-to guide or tutorial. It contains the following sections: + +1. Assembly +2. Parts +3. Vectors +4. Primers +5. Cache blocks + +# Assembly + +## Special overhangs: GTCT and CGAG + +Both `GTCT` and `CGAG` are special overhangs that make dd assembly different than alternative GoldenGate standards like MoClo. These two sites enable recursive GoldenGate assembly of genetic parts by containing part of the BsaI cut site, but not an edge base pair. This edge base pair can be methylated, preventing cutting during a GoldenGate assembly. This methylation is then deprotected during amplification so that BsaI can be used again for another assembly reaction. Since these overhangs are within the BsaI cut site, when BsaI is used in subsequent reactions, it cuts user-defined DNA. Let's take a detailed look at what a vector would look like. + +``` +>recursive vector +... g[GTCT](NGAGACC---GGTCTCN)[CGAG]ACc ... + +>insert +GGTCTCN [GTCT]CA (NNNN --- NNNN) [CGAG] NGAGACC + +>result +g[GTCT]CA (NNNN --- NNNN) [CGAG]ACc +``` + +To break the `recursive` vector down: +1. `GGTCT` is BsaI in the forward direction and `GAGACC` is BsaI in the reverse direction. It cuts `1,4`, or `GGTCTC N [NNNN]` where `NNNN` is the overhang. +2. The rest of the circular vector (ori and amp) are simplified with `...` +3. The lowercase letters are methylated cytosines (in the complement for `g`) +4. The two overhangs in brackets are our special overhangs `GTCT` and `CGAG` +5. The sequence within the parathesis is sequence that will get cut out during the GoldenGate reaction, and will be replaced with our sequence of interest. + +To break the insert down: +1. `GGTCTC` and `GAGACC` is still BsaI +2. The user insert is `(NNNN --- NNNN)`. The 4 NNNNs on both sides are the overhangs which will be exposed after methylation is removed. +3. `CA` after `[GTCT]` is spacer needed to properly space the user DNA from the methylation-exposed BsaI cut sites. + +In dd assembly, there are two kinds of vectors: `recursive` vectors and `base` vectors. `recursive` vectors are just like they are described above, while `base` vectors derive their overhangs from `linkers` or from a special kind of insert called a `replaceable`, creating a `replaceable` vector. Since `base` vectors by themselves do not contain more BsaI sites, they can also be used to create `shuttle` vectors. We will go over each before diving into specific overhangs for parts. Briefly: + +1. `recursive` vectors are used for creating synthetic DNA, independent of partification. +2. `linkers` are used to create multigene constructs. **Does not require vector intermediates** +3. `replaceable` vectors are created from `base` vectors by inserting genetic parts and a replacement insert. Mostly just used for expression vectors, but can also be used for shuttle vectors. + +## Recursive construction + ## Linkers The most basic dd assembly will create a transcriptional unit. Oftentimes, however, you will want to be able to combine different transcriptional units together. In order to do these multi-level assemblies, we use linkers. Linkers are genetic parts that are used within an assembly reaction that define the part overhangs of the assembled construct, for use in the next assembly reaction. For example, we could have 3 genetic parts: @@ -37,4 +89,54 @@ The most basic dd assembly will create a transcriptional unit. Oftentimes, howev * GFP * Terminator -While we could construct a simple transcriptional unit of `["Promoter+RBS", "GFP", "Terminator"]`, we might want to use this whole transcriptional unit in a different construct. In order to do that, we will add 2 linkers. The prefix linker (A) +While we could construct a simple transcriptional unit of `["Promoter+RBS", "GFP", "Terminator"]`, we might want to use this whole transcriptional unit in a different construct. In order to do that, we will add 2 linkers. The prefix linker (A) and the suffix linker (B). These linkers have additional numbers, + +## Replaceable vectors + +# Parts + +## Orthogonal primer binding sites + +### CDS fusion using SapI +``` +NNN TGA AGAGC ACTT +``` + +In dd assembly, CDSs have either their protein tags directly fused to them, or use SapI fusion. SapI fusions are enabled by the following observation: You can overlap SapI with a stop codon to specifically cut the last codon of a protein. By cutting the last codon, without cutting any other sequence, we can create seamless protein fusions for any protein. Proteins do not need to be specifically designed to have fusion tags - + +# Vectors + +## Standard oriT +dd assembly takes is opinionated in how transfer to non-cloning organisms should be done. + +# Cache blocks + +Cache blocking is a concept unique to dd assembly, designed to give us a way to feasibly synthesize and test massive sequences. It stems from one fundamental observation: we are limited in our ability to synthesize correct DNA. Cache blocking aims to minimize the necessity of synthesis when creating and testing DNA. + +In practical terms, you chunk a given sequence into `cache blocks`, which are clonally verified. Each `cache block` is defined by dd assembly overhangs, so can be used like any other construct, but unlike when doing classic dd assembly, these `cache blocks` are seamless. They are fragmented at dd assembly overhangs, but they do not have scar sequences. Subsequent cache block assemblies simply maintain whatever overhangs were on the edge `cache blocks`, and these assemblies can create new `cache blocks` that get sequence verified, or create a final desired sequence. + +## Mutational limiting + +When you use directly utilize synthetic DNA, you are typically limited by the mutation rate of the synthesis reaction. Synthesis blocks have a lot of mutations. This means you have to clone more intermediate fragments. One idea behind `cache blocks` is that, if you take the upfront cost of clonally verifying each block, the replacement of any given block within a target sequence becomes increasingly lower. + +Let's say, for example, we have a 8kbp metabolic circuit that is `cache block`ed down to 250bp, or 32 `cache blocks`. If you use an AI system to mutate one of the genes, or perhaps some of the ribosomal binding sites, you would just swap the specific blocks you need to change. If you're only changing a few blocks, you only need to synthesize the 250bp from those blocks, lowering synthesis costs, but also lowering mutation rates to a screenable level, whereas it is very difficult to do that with 8kbp of purely synthetic DNA. + +An important thing to note here is that the `cache blocks` essentially act as constants: so the same concept that works with 8kbp works with 50kbp, or perhaps even **genomes**. In this way, we make whole genome rearrangement and testing trivial: you only need to resynthesize the specific blocks you're changing. The entire system can work computationally in a consistent manner for testing any piece of DNA in a modular fashion. + +## Assembly caching + +An astute reader may still observe that we need to assemble an awful lot of `cache blocks`. There are two ways we handle this: assembly caching using `identity` linkers and clone-less assembly. + +Linker based recursion is similar to the above linker section with one exception: all linkers are `identity` linkers. `identity` linkers do not redefine their overhangs. They simply take in `cache blocks` (or parts, since they appear the same to dd assembly), and spit out assemblies of those `cache blocks` with the edge overhangs exposed. This enables you to create compositions of `cache blocks` as `cache blocks` themselves - for example, in our 8bkp example, if we are only changing the first 2kbp, we can `cache block` the remaining 6kbp as a single block. This new block can be sequence verified and used further. + +However, we can also use clone-less assembly, as described above. Basically, we do not pause for a cloning step between putting together a number of `cache blocks` together. We simply amplify the resultant GoldenGate and continue with the next step of assembly. The most important thing that this process does is limit our need to achieve equimolar GoldenGate ratios for efficient assembly - as this would take in intermediate quantification and normalization step which is, ironically, more expensive and more annoying than simply recursing on assemblies. This also allows reuse of overhangs - for example, if you want to assemble a transcriptional unit. + +## Plasmid resynthesis and Genome testing + +There are two killer applications of `cache blocks`: plasmid resynthesis and genome testing. Many plasmids use the same components - ampicillin resistance, pUC origin, etc - and these can be cached in such a way that users can synthesize arbitrary plasmids without even thinking about parts - in each case, from an end-user perspective, the only thing that gets synthesized is the user-specified DNA with some minimal flanking sequence to compensate on either side since the mutations are limited and we assembly cache most of the vector backbone. In scaled facilities that can handle large quantities of DNA, this eliminates the need for any vector onboarding. + +Perhaps the most difficult part of creating new synthetic genomes is testing whether or not changes work. Once cached, however, this becomes much much easier: parts can be swapped out piecewise, with synthesis and assembly only occuring at locations with changes, in a similar way to plasmid resynthesis, except at scale. Testing any particular change becomes just a task of assembly. + +### Genome replacement + +Rather than focus on methods of genome replacement that are specific to certain organisms on the basis of their unique properties (for example, homologous recombination/integration into yeast, natural competence of Bacillus subtilis), dd assembly foucses