| 质粒名称: | pUC-GFP-AT |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 133306 |
| 存储实验室: | Chris Barnes |
|
载体骨架基本信息 |
|
| 载体名称: | pSF_UC_OXB20-daGFP |
| 载体出品公司: | Oxford Genetics |
| 空载体大小: | 4700 bp |
| 完整质粒大小: | 6475 bp |
| 载体修饰: | 原始SC101 ori替换为pUC ori |
| 载体类型: | 细菌表达，合成生物学 |
| 载体抗性: | 卡那霉素50 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | axe/txe toxin-antitoxin cassette |
| 物种来源: | E. coli |
| 插入大小: | 1998 bp |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | SacI（如果已销毁，未知） |
| 3’克隆位点: | XhoI（如果已销毁，未知） |
| 5’测序引物 | TCGTTGCGTTACACACAC |
| 3’测序引物 | AGTCAGTCAGTGCAGGAG |
|
文献引用方法 |
|
| 材料和方法部分: | pUC-GFP-AT was a gift from Chris Barnes (Addgene plasmid # 133306 ; http://n2t.net/addgene:133306 ; RRID:Addgene_133306) |
| 参考文献部分: | Two New Plasmid Post-segregational Killing Mechanisms for the Implementation of Synthetic Gene Networks in Escherichia coli. Fedorec AJH, Ozdemir T, Doshi A, Ho YK, Rosa L, Rutter J, Velazquez O, Pinheiro VB, Danino T, Barnes CP. iScience. 2019 Apr 26;14:323-334. doi: 10.1016/j.isci.2019.03.019. Epub 2019 Mar 22. 10.1016/j.isci.2019.03.019 PubMed 30954530 |
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 pUC-GFP-AT 6475 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION ""Burdensome"" GFP expressing plasmid carrying axe/txe
toxin-antitoxin for plasmid stabilisation.
ACCESSION .
VERSION .
KEYWORDS pUC-GFP-AT
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6475)
AUTHORS Fedorec AJH, Ozdemir T, Doshi A, Ho YK, Rosa L, Rutter J, Velazquez
O, Pinheiro VB, Danino T, Barnes CP
TITLE Two New Plasmid Post-segregational Killing Mechanisms for the
Implementation of Synthetic Gene Networks in Escherichia coli.
JOURNAL iScience. 2019 Apr 26;14:323-334. doi: 10.1016/j.isci.2019.03.019.
Epub 2019 Mar 22.
PUBMED 30954530
REFERENCE 2 (bases 1 to 6475)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6475
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 58..129
/label=5' beta-globin insulator
/note="insulator upstream of the human beta-globin locus
(Farrell et al., 2002)"
terminator 134..180
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
polyA_signal 198..246
/note="synthetic polyadenylation signal"
polyA_signal 2253..2374
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2264..2283)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 2318..2337
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
terminator 2489..2535
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
terminator 2565..2701
/label=rrnG terminator
/note="transcription terminator from the E. coli ribosomal
RNA rrnG operon (Albrechtsen et al., 1991)"
misc_feature 2746..2817
/label=3' beta-globin insulator
/note="insulator downstream of the human beta-globin locus
(Farrell et al., 2002)"
rep_origin complement(3043..3631)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3123..3142)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
promoter 4251..4319
/label=recA (Delta-LexA) promoter
/note="constitutively active E. coli recA promoter lacking
the LexA binding site (Brent and Ptashne, 1981)"
CDS 4336..5046
/codon_start=1
/product="green fluorescent protein with peak excitation at
510 nm and peak emission at 521 nm"
/label=daGFP
/translation="MTALTEGAKLFEKEIPYITELEGDVEGMKFIIKGEGTGDATTGTI
KAKYICTTGDLPVPWATLVSTLSYGVQCFAKYPSHIKDFFKSAMPEGYTQERTISFEGD
GVYKTRAMVTYERGSIYNRVTLTGENFKKDGHILRKNVAFQCPPSILYILPDTVNNGIR
VEFNQAYDIEGVTEKLVTKCSQMNRPLAGSAAVHIPRYHHITYHTKLSKDRDERRDHMC
LVEVVKAVDLDTYQ"
terminator 5053..5188
/label=rrnG terminator
/note="transcription terminator from the E. coli ribosomal
RNA rrnG operon (Albrechtsen et al., 1991)"
CDS 5445..6239
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(5499..5518)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 6109..6128
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
terminator 6252..6388
/label=rrnG terminator
/note="transcription terminator from the E. coli ribosomal
RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
1 atcgaatctc gtcagctatc gtcagcttac ctttttggca gcgatcgcgg ctcccgacat
61 cttggaccat tagctccaca ggtatcttct tccctctagt ggtcataaca gcagcttcag
121 ctacctctca attcaaaaaa cccctcaaga cccgtttaga ggccccaagg ggttatgcta
181 tcaatcgttg cgttacacac acaaaaaacc aacacacatc catcttcgat ggatagcgat
241 tttattatct aactgctgat cgagtgtagc cagatctttg tcgatcctac catccactcg
301 acacacccgc cagcggccgc tgccaagctt ccgagctcga tacttctttc aaattcaagt
361 taaatttata ttccattact tctcctccaa tatataaaaa ttacttaact gtatccaaat
421 ttaatatgta aataaatcat taatttttag ctaaattcta tgagaagttt atttttgttg
481 tttttgctaa ttgactaaaa ctagtacaaa aaatttaaaa tagattatta aacctattgc
541 ttattctatt cagtcatata tataagcgtc atcctttagt aactaacatc ccacactaca
601 atagtattat aacatatcta ttcaacttca aattgttgaa atttaagcca tttatattta
661 cttttcctca actttagaaa tgattttact aaccttatat ccgtaatacg cgtaacaaac
721 acattcatta ataacaatca acactaagct atattagctt tttaaatgac tagggtaaat
781 aaaataagga tcaaaaactt ttaagtttct gaccctttcc ttacttccga ttggttaata
841 gtgatctttt gcagaataaa taaatatcgt ttcattttca actctatata tcagtctatg
901 ttcatctgta attcttctgg accattttcc agataaatca tgctttaatg gctcaggttt
961 tcctaatcca gcaaaggggg aacgatcgat atcttttatt aacttgttaa tcttttttat
1021 attgcttttg tttccttgct catgccaata aagataatca tcccaagcat catcagacca
1081 agccttaatc atcagattca acctcgatta agtcatgtgt tttaaatgca cctttggaga
1141 attgttcatc tcctcgacga attttttcca tgacgtaatt attagaaagt gttctcaacg
1201 tttcttgcat agaatcataa tctctttttg ataatacaac aactgtatct tctacatctt
1261 tacttgttac aataagtgtt tcagcatcct cattaacttg tttcatataa ctacgtaaat
1321 tttggcggaa atttgaataa gctactgctt ccattccttt caccccttat ttatttctat
1381 aaaacaattg tacattatat tgtacaatta agcaaatgat taatattcta acttcttata
1441 attaaaagtg ctatcacttt tagtttttag gataaaaggt gacgtcactt tttgagtgct
1501 ggtacctcaa atcctgcttt ttcttgggca atcaataaat ctaacgcaat acacataatc
1561 ctggattcac ttttgccgta cttctgcttt agttgctgca attctttttg tctcttttca
1621 tccaacgtaa cgggtaaccg tttatttcct attgcaattg gcattagtcg cttatggatc
1681 ctgaacttga ccgtaaacga atgccatatc aacgatttga gaggaaaaaa ggcaataatc
1741 atttataccg aaaagtgaga atgtgtgaaa taaggacaaa aaacctagta actaagggaa
1801 tggttcttat ttcgcaccaa tcgctccaac cgcccccgcc gcttaggtgg tgcgaaataa
1861 gcgggcaaca cccgataacc taagagaaag aaggagatcc atatgaataa aatcattcct
1921 ctattgtcag tcctatttgt tggcgaaata cttatgcgcc atgctgcagc tgtgcattta
1981 aacttcgtag tatttttggc aatactggta ttttgtttgg tcttttggcg ctttgtatta
2041 gcccttgtga gtctactatt caagatgatt gcagccatat ttttcatagg gttatttcga
2101 tgaaaggaga gagtatcgga tcccccgggc tcgaggtttc tagaagttgt ctcctcctgc
2161 actgactgac tgatacaatc gatttctgga tccgcaggcc tctgctagct tgactgactg
2221 agatacagcg taccttcagc tcacagacat gataagatac attgatgagt ttggacaaac
2281 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
2341 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
2401 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
2461 tggtattggc ccatctctat cggtatcgta gcataacccc ttggggcctc taaacgggtc
2521 ttgaggggtt ttttgtgccc ctcgggccgg attgctatct accggcattg gcgcagaaaa
2581 aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga
2641 tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta
2701 aggtcgtcag ctatcctgca ggcgatctct cgatttcgat caagacattc ctttaatggt
2761 cttttctgga caccactagg ggtcagaagt agttcatcaa actttcttcc ctccctaatc
2821 tcattggtta ccttgggcta tcgaaactta attaaccagt caagtcagct acttggcgag
2881 atcgacttgt ctgggtttcg actacgctca gaattgcgtc agtcaagttc gatctggtcc
2941 ttgctattgc acccgttctc cgattacgag tttcatttaa atcatgtgag caaaaggcca
3001 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
3061 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
3121 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
3181 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
3241 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
3301 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
3361 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
3421 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
3481 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
3541 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
3601 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
3661 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
3721 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
3781 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
3841 ctgtctattt cgttcatcca tagttgcatt taaatttccg aactctccaa ggccctcgtc
3901 ggaaaatctt caaacctttc gtccgatcca tcttgcaggc tacctctcga acgaactatc
3961 gcaagtctct tggccggcct tgcgccttgg ctattgcttg gcagcgccta tcgccaggta
4021 ttactccaat cccgaatatc cgagatcggg atcacccgag agaagttcaa cctacatcct
4081 caatcccgat ctatccgaga tccgaggaat atcgaaatcg gggcgcgcca agctgttgtg
4141 accgcttgct ctagccagct atcgagttgt gaaccgatcc atctagcaat tggtctcgat
4201 ctagcgatag gcttcgatct agctatgtag aaacgccgtg tgctcgatcg cttgataagg
4261 tccacgtagc tgctataatt gcttcaacag aacatattga ctatccggta ttacccggca
4321 aaggaggtag ccaacatgac agctctgaca gagggagcga aactgttcga gaaggagatc
4381 ccgtacatca cagagttgga aggagatgtc gaagggatga aattcatcat caagggagaa
4441 ggaacgggtg acgcaacgac cggaacgatc aaggccaaat acatttgtac cacgggcgat
4501 ttgcctgtcc cctgggcgac gcttgtaagc acgctctcgt atggtgtcca gtgcttcgcg
4561 aaatatccat cgcacattaa ggactttttc aagtcggcca tgccagaagg ttacacacaa
4621 gaacgaacca tctcctttga gggggacgga gtgtataaga cacgcgcgat ggtaacgtac
4681 gagcgcgggt ccatctacaa cagggtaact cttactgggg agaactttaa gaaggacggg
4741 catatcttgc ggaaaaacgt ggcatttcaa tgtccgccct cgattctgta tattctcccg
4801 gacacggtga ataatgggat cagagtggag ttcaaccagg catacgatat tgagggtgtg
4861 actgaaaagc tcgtcaccaa atgcagccag atgaatcgcc cccttgcggg atcagcagcc
4921 gtccatatcc cccggtatca ccatattacc taccacacaa aactctcaaa agacagagat
4981 gagagaaggg accatatgtg cctggtcgaa gtagtgaagg cggtggatct tgatacatac
5041 cagtaggcta gccattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact
5101 cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt
5161 gtcctcctta ccagaaattt atccttaagg gcgcgcctgg tgtaccgaga acgatcctct
5221 cagtgcgagt ctcgacgatc catatcgttg cttggcagtc agccagtcgg aatccagctt
5281 gggacccagg aagtccaatc gtcagatatt gtactcaagc ctggtcacgg cagcgtaccg
5341 atctgtttaa acctagatat tgatagtctg atcggtcaac gtataatcga gtcctagctt
5401 ttgcaaacat ctatcaagag acaggatcag caggaggctt tcgcatgatt gaacaagatg
5461 gattgcacgc aggttctccg gcggcttggg tggagaggct attcggctat gactgggcac
5521 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgtccgg
5581 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
5641 ggctatcgtg gctggcgacg acgggcgttc cttgcgcggc tgtgctcgac gttgtcactg
5701 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
5761 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
5821 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
5881 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
5941 cgccagccga actgttcgcc aggctcaagg cgtctatgcc cgacggcgag gatctcgtcg
6001 tgacccacgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
6061 tcatcgactg tggccgtctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
6121 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttccttgtg ctttacggta
6181 tcgccgcgcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgac
6241 cgattctagg tgcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact
6301 cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt
6361 gtcctcctta ccagaaattt atccttaagg tcgtttaaac tcgactctgg ctctatcgaa
6421 tctccgtcgt ttcgagctta cgcgaacagc cgtggcgctc atttgctcgt cgggc
//
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