| 质粒名称: | pTW084b |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 116894 |
| 存储实验室: | David Liu |
|
载体骨架基本信息 |
|
| 载体名称: | SC101 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | 5437 bp |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 低拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | g3N |
| 物种来源: | Synthetic |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pTW084b was a gift from David Liu (Addgene plasmid # 116894 ; http://n2t.net/addgene:116894 ; RRID:Addgene_116894) |
| 参考文献部分: | Continuous directed evolution of proteins with improved soluble expression. Wang T, Badran AH, Huang TP, Liu DR. Nat Chem Biol. 2018 Aug 20. pii: 10.1038/s41589-018-0121-5. doi: 10.1038/s41589-018-0121-5. 10.1038/s41589-018-0121-5 [pii] PubMed 30127387 |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pTW084b 5437 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Activity-independent SE-PACE AP.
ACCESSION .
VERSION .
KEYWORDS pTW084b
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5437)
AUTHORS Wang T, Badran AH, Huang TP, Liu DR
TITLE Continuous directed evolution of proteins with improved soluble
expression.
JOURNAL Nat Chem Biol. 2018 Aug 20. pii: 10.1038/s41589-018-0121-5. doi:
10.1038/s41589-018-0121-5.
PUBMED 30127387
REFERENCE 2 (bases 1 to 5437)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5437
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS complement(1..951)
/codon_start=1
/gene="rep101"
/product="RepA protein needed for replication with the
pSC101 origin"
/label=Rep101
/translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFESKF
LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
rep_origin 999..1221
/label=pSC101 ori
/note="low-copy replication origin that requires the Rep101
protein"
CDS complement(3090..3113)
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
terminator 3945..3972
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator 4064..4150
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind 4303..4320
/label=pBAD Reverse
/note="For vectors with E. coli araBAD promoter, reverse
primer"
CDS complement(4366..5226)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 4989..5008
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5227..5331)
/gene="bla"
/label=AmpR promoter
primer_bind 5399..5417
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
ORIGIN
1 tcagatcctt ccgtatttag ccagtatgtt ctctagtgtg gttcgttgtt tttgcgtgag
61 ccatgagaac gaaccattga gatcatgctt actttgcatg tcactcaaaa attttgactc
121 aaaactggtg agctgaattt ttgcagttaa agcatcgtgt agtgtttttc ttagtccgtt
181 acgtaggtag gaatctgatg taatggttgt tggtattttg tcaccattca tttttatctg
241 gttgttctca agttcggtta cgagatccat ttgtctatct agttcaactt ggaaaatcaa
301 cgtatcagtc gggcggcctc gcttatcaac caccaatttc atattgctgt aagtgtttaa
361 atctttactt attggtttca aaacccattg gttaagcctt ttaaactcat ggtagttatt
421 ttcaagcatt aacatgaact taaattcatc aaggctaatc tctatatttg ccttgtgagt
481 tttcttttgt gttagttctt ttaataacca ctcataaatc ctcatagagt atttgttttc
541 aaaagactta acatgttcca gattatattt tatgaatttt tttaactgga aaagataagg
601 caatatctct tcactaaaaa ctaattctaa tttttcgctt gagaacttgg catagtttgt
661 ccactggaaa atctcaaagc ctttaaccaa aggattcctg atttccacag ttctcgtcat
721 cagctctctg gttgctttag ctaatacacc ataagcattt tccctactga tgttcatcat
781 ctgagcgtat tggttataag tgaacgatac cgtccgttct ttccttgtag ggttttcaat
841 cgtggggttg agtagtgcca cacagcataa aattagcttg gtttcatgct ccgttaagtc
901 atagcgacta atcgctagtt catttgcttt gaaaacaact aattcagaca tacatctcaa
961 ttggtctagg taattttaat cactatacca attgagatgg gctagtcaat gataattact
1021 agtccttttc ctttgagttg tgggtatctg taaattctgc tagacctttg ctggaaaact
1081 tgtaaattct gctagaccct ctgtaaattc cgctagacct ttgtgtgttt tttttgttta
1141 tattaaagtg gttataattt atagaataaa gaaagaataa aaaaagataa aaagaataga
1201 tcccagccct gtgtataact cactacttta gtcagttccg cagtattaca aaaggatgtc
1261 gcaaacgctg tttgctcctc tacaaaacag accttaaaac cctaaaggct taagtagcac
1321 cctcgcaagc tcgggcaaat cgctgaatat tccttttgtc tccgaccatc aggcacctga
1381 gtcgctgtct ttttcgtgac attcagttcg ctgcgctcac ggctctggca gtgaatgggg
1441 gtaaatggca ctacaggcgc cttttatgga ttcatgcaag gaaactaccc ataatacaag
1501 aaaagcccgt cacgggcttc tcagggcgtt ttatggcggg tctgctatgt ggtgctatct
1561 gactttttgc tgttcagcag ttcctgccct ctgcttttcc agtctgacca cttcggctta
1621 tcccgtgaca ggtcattcag actggctaat gcacccagta aggcagcggg ctcatcaaca
1681 ggcttacccg tcgtacggtc aagaggacat ccgggcaaat gcaacgaaag gctcagtcga
1741 aagactgggc ctttcgttgc gctgcggaga cttagggacc ctaaaacaag atacattgta
1801 tgaaaataca agaaagtttg ttcattaggc accccgggct ttactcgtaa agcttccggc
1861 gcgtatgttg tgtcgaccga cctgcaggtg cagtaaaaaa ggaaaaaaat gaaaaaatta
1921 ttattcgcaa ttcctttatg tctcagctac gaaaccgaaa tcttgaccgt cgaatatggt
1981 ctgctgccaa tcggcaagat tgttgaaaaa cgtattgaat gtacggtcta ctcagtggat
2041 aacaacggca atatctacac ccagccggtg gcccagtggc atgaccgtgg tgaacaggaa
2101 gtgttcgaat attgtctgga agacggatct ttaatccgtg ccacaaagga tcacaaattt
2161 atgactgtag atggtcagat gctcccaatc gacgaaattt ttgaacgcga attagacctg
2221 atgcgcgtgg ataatctccc gaattaaacg gagccaatgt acgcaaaaaa ccccgcttca
2281 gcggggtttt ttcgccaaaa aaaaccccgc cctgtcaggg gcggggtttt tttttagggc
2341 cctactgact gttagctgga gaccgtgaca agagtgccct gcccccagta gtcaaaaagt
2401 ccggtgacac agtagtataa cgccgtatca tccgagcgaa ctttgctcat ttgcaaatag
2461 acagtgtttt cgcaatcgtc tttgctgatg ataaagcgat ccttcaaggc actgttataa
2521 tcagtggttc cgtcgcccca gatgacgcca atccactcca agccacgccc cggtgcctgg
2581 cgtacccaat tgaccccata atccgtcaac gagaaaccac tcacagcgca gctcagcttt
2641 aacgagcctc caggctgcac aagaccacca ccactttcaa gaagcttaac ctcactccct
2701 ccacccgaag accctccgcc gcctgaaccc ccacctccgg agccaccacc cccgcgttta
2761 agttcaacct tcgtgccctg accgaagacc caatggttgg agtaccacaa tgcacaatag
2821 tatgtagcga aatcctctgg ttgcagactg ctgatagtaa gtgtggcctt gtcaccaatc
2881 aaactccccg aaaaacggct agggacgccc ggtgcgcggt tattagtacc gccaatcaaa
2941 cccttgaaca acttacccgg tttttcctgg acccacgaag cgtaattact ggttgtgact
3001 gccccagtgc tcgaacggca tgtaattgta acgcgatctc ccactgacgc tgataagctt
3061 gatggcgact gcgtcataac aatgtcgcgc ttgtcatcgt cgtccttgta gtctacgaat
3121 tttaccctcg cttccacgac aacaccgata atcttgcagt ttccgttgat aggagtcata
3181 ggccatgaag gattcaggcc tttcaggtac ttctgaccgc catctatgac cagtttcttg
3241 aatgttgctt cgttcgcgtc agtcagtttg gctacaacaa ggcttccatt cactggctcc
3301 cgtccagtat ctactaacac catatgacct tcagggatgc tttgacctac aggtgaggtc
3361 atggaatcac cttcaacctt cagccagaat ccattgccta ataagttaac gtcactgtca
3421 taccattcat caatgtcctt gatatcgtag ggttcacaag cttcacacca cgaacgagct
3481 ctaaccatgc taatcaatgg atatttccct ttgggctcaa cgtgcccaac aaatctaaca
3541 ttcgaatcag aggtgccatt gagcagccag tcaacactta cgccaagagc tgacgcaagt
3601 tctggtaaaa agcgtggtcg cttagtttta ccgttttcga gctgctctat agactgctgg
3661 gtagtcccca ccttttgagc aagttcagcc tggttaagtc caagctgaat tcttttgctt
3721 tttaccctgc tgctgatact catttttttt tcctccttac tctagtaaaa gttaaacaaa
3781 attatttgta gagggaaacc gttgtggtct ccctgaatat agataccgag ccttatgcat
3841 gcccgtaaag ttatccagca accactcata gacctagggc agcagatagg gacgacgtgg
3901 tgttagctgt gctctagaat cctcagcgag tttgtagaaa cgcaaaaagg acatccgtca
3961 ggatggcctt ctgcttaatt tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc
4021 ctccgggccg ttgcttcgca acgttcaaat ccgctcccgg cggatttgtc ctactcagga
4081 gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtctttcgac tgagcctttc
4141 gttttatttg atgcctggca gttccctact ctcgcatggg gagaccccac actaccatcg
4201 gcgctacggc gtttcacttc tgagttcggc atggggtcag gtgggaccac cgcgctactg
4261 ccgccaggca aattctgttt tatcagaccg cttctgcgtt ctgatttaat ctgtatcagg
4321 ctgaaaatct tctctcatcc gccaaaacag ccatgcccat ggcgtttacc aatgcttaat
4381 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
4441 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
4501 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
4561 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
4621 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
4681 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
4741 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
4801 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
4861 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
4921 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccagcgtc
4981 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
5041 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
5101 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
5161 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
5221 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
5281 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
5341 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
5401 taggcgtatc acgaggccct taggccaaat aggccgt
//
