| 质粒名称: | pFE-sfGFP |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 126575 |
| 存储实验室: | Jerome Bonnet |
|
载体骨架基本信息 |
|
| 载体名称: | pBEST-OR2-OR1-Pr-UTR1-deGFP-T500 |
| 完整质粒大小: | 3611bp |
| 载体类型: | 合成生物学;无细胞蛋白合成 |
| 筛选标记: | 新霉素（G418） |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 30 ℃ |
| 宿主菌: | KL740+ |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | sfGFP |
| 物种来源: | Synthetic |
| 插入大小: | 720 bp |
|
文献引用方法 |
|
| 材料和方法部分: |
pBEAST2-sfGFP was a gift from Jerome Bonnet (Addgene plasmid # 126575 ; http://n2t.net/addgene:126575 ; RRID:Addgene_126575)
|
| 参考文献部分: | Plug-and-play metabolic transducers expand the chemical detection space of cell-free biosensors. Voyvodic PL, Pandi A, Koch M, Conejero I, Valjent E, Courtet P, Renard E, Faulon JL, Bonnet J. Nat Commun. 2019 Apr 12;10(1):1697. doi: 10.1038/s41467-019-09722-9. 10.1038/s41467-019-09722-9 PubMed 30979906 |
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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 pBEAST2-sfGFP 3611 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Strong constitutive expression of super-folder GFP for cell-free
expression.
ACCESSION .
VERSION .
KEYWORDS pBEAST2-sfGFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3611)
AUTHORS Voyvodic PL, Pandi A, Koch M, Conejero I, Valjent E, Courtet P,
Renard E, Faulon JL, Bonnet J
TITLE Plug-and-play metabolic transducers expand the chemical detection
space of cell-free biosensors.
JOURNAL Nat Commun. 2019 Apr 12;10(1):1697. doi: 10.1038/s41467-019-09722-9.
PUBMED 30979906
REFERENCE 2 (bases 1 to 3611)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..3611
/organism="synthetic DNA construct"
/mol_type="other DNA"
terminator 148..175
/label=T7Te terminator
/note="phage T7 early transcription terminator"
RBS 343..365
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 370..1083
/codon_start=1
/product="GFP variant that folds robustly even when fused
to poorly folded proteins (Pedelacq et al., 2006)"
/label=superfolder GFP
/translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
FVTAAGITHGMDELYK"
primer_bind complement(1426..1443)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1597..2185)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1677..1696)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2356..3216)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 2979..2998
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(3217..3321)
/gene="bla"
/label=AmpR promoter
primer_bind 3389..3407
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
ORIGIN
1 cgcctgatgc gtgaacgtga cggacgtaac caccgcgaca tgtgtgtgct gttccgctgg
61 ctcggatacc cttactctgt tgaaaacgaa tagataggtt aaggaacggt tatttctgcg
121 tagatctatc ttacacagca tcacactggc tcaccttcgg gtgggccttt ctgcgtttat
181 atactagaga gagaatataa aaagccagat tattaatccg gcttttttat tatttaggca
241 actgaaacga ttcggatcct gtattactat tcttatgagc taacaccgtg cgtgttgaca
301 attttacctc tggcggtgat aatggttgca gctagcaata attttgttta actttaagaa
361 ggagatataa tgcgtaaagg cgaagagctg ttcactggtg tcgtccctat tctggtggaa
421 ctggatggtg atgtcaacgg tcataagttt tccgtgcgtg gcgagggtga aggtgacgca
481 actaatggta aactgacgct gaagttcatc tgtactactg gtaaactgcc ggtaccttgg
541 ccgactctgg taacgacgct gacttatggt gttcagtgct ttgctcgtta tccggaccat
601 atgaagcagc atgacttctt caagtccgcc atgccggaag gctatgtgca ggaacgcacg
661 atttccttta aggatgacgg cacgtacaaa acgcgtgcgg aagtgaaatt tgaaggcgat
721 accctggtaa accgcattga gctgaaaggc attgacttta aagaagacgg caatatcctg
781 ggccataagc tggaatacaa ttttaacagc cacaatgttt acatcaccgc cgataaacaa
841 aaaaatggca ttaaagcgaa ttttaaaatt cgccacaacg tggaggatgg cagcgtgcag
901 ctggctgatc actaccagca aaacactcca atcggtgatg gtcctgttct gctgccagac
961 aatcactatc tgagcacgca aagcgttctg tctaaagatc cgaacgagaa acgcgatcat
1021 atggttctgc tggagttcgt aaccgcagcg ggcatcacgc atggtatgga tgaactgtac
1081 aaatgatgaa ctttatctga gaatagtcaa tcttcggaaa tcccaggtgg catgctaaaa
1141 gtctcgtaaa gcgttctatc aataacccgt tggtgcaaag cccgccgaaa ggcgggcttt
1201 tctgtccgtc tcagaatcgg ccgtgaacaa taaaatagtt tcggtattat tgaccacttc
1261 cgagtagaat cgtgcttcag taagagtcga ccgatgccct tgagagcctt caacccagtc
1321 agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac tgtcttcttt
1381 atcatgcaac tcgtaggaca ggtgccggca gcgctcttcc gcttcctcgc tcactgactc
1441 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
1501 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
1561 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
1621 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
1681 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
1741 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
1801 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
1861 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
1921 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
1981 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
2041 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
2101 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
2161 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
2221 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
2281 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
2341 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
2401 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
2461 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
2521 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
2581 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
2641 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
2701 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
2761 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
2821 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
2881 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
2941 gcggcgaccg agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag
3001 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
3061 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
3121 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
3181 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
3241 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
3301 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
3361 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca
3421 agaattctgg cgaatcctct gaccagccag aaaacgacct ttctgtggtg aaaccggatg
3481 ctgcaattca gagcgccagc aagtggggga cagcagaaga cctgaccgcc gcagagtgga
3541 tgtttgacat ggtgaagact atcgcaccat cagccagaaa accgaatttt gctgggtggg
3601 ctaacgatat c
//
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