质粒名称: | pProm1_BCD1-GFP |
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出品公司: | Addgene |
目录编号: | 80649 |
存储实验室: | Nathan Hillson |
载体骨架基本信息 |
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载体名称: | N/A |
空载体大小: | 2353bp |
完整质粒大小: | 3070 bp |
载体类型: | p15A origin, constitutive promoter, KanR |
筛选标记: | Kanamycin |
载体抗性: | Kanamycin |
生长温度: | 37 ℃ |
宿主菌: | DH10B |
拷贝数: | 中拷贝 |
插入基因信息 |
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插入基因名称: | GFP / fGFP |
插入大小: | 717bp |
基因突变: | NONE |
启动子: | constitutive promoter |
克隆方法: | 未知 |
5’测序引物 | QB3284_Fwd CGATCCTCATCCTGTCTCTTGATC |
3’测序引物 | QB3810_Rev CGAGCGTAGCGAGTCAGTG |
文献引用方法 |
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材料和方法部分: | pProm1_BCD1-GFP was a gift from Nathan Hillson (Addgene plasmid # 80649 ; http://n2t.net/addgene:80649 ; RRID:Addgene_80649) |
参考文献部分: | PR-PR: cross-platform laboratory automation system. Linshiz G, Stawski N, Goyal G, Bi C, Poust S, Sharma M, Mutalik V, Keasling JD, Hillson NJ. ACS Synth Biol. 2014 Aug 15;3(8):515-24. doi: 10.1021/sb4001728. Epub 2014 Jan 14. 10.1021/sb4001728 PubMed 25126893 |
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LOCUS pProm1_BCD1-GFP 3070 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Promoter1 with BCD1-gfp. ACCESSION . VERSION . KEYWORDS pProm1_BCD1-GFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3070) AUTHORS Linshiz G, Stawski N, Goyal G, Bi C, Poust S, Sharma M, Mutalik V, Keasling JD, Hillson NJ TITLE PR-PR: cross-platform laboratory automation system. JOURNAL ACS Synth Biol. 2014 Aug 15;3(8):515-24. doi: 10.1021/sb4001728. Epub 2014 Jan 14. PUBMED 25126893 REFERENCE 2 (bases 1 to 3070) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..3070 /organism="synthetic DNA construct" /mol_type="other DNA" terminator 97..191 /label=lambda t0 terminator /note="transcription terminator from phage lambda" CDS complement(222..1016) /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 GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(333..352) /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind 943..962 /label=Neo-R /note="Neomycin resistance gene, reverse primer" CDS 1468..2181 /codon_start=1 /product="GFP variant that folds robustly even when fused to poorly folded proteins (Pedelacq et al., 2006)" /label=superfolder GFP /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE FVTAAGITHGMDELYK" primer_bind complement(1498..1526) /label=GFP-R /note="GFP, reverse primer. Does NOT anneal to EGFP" primer_bind 2121..2141 /label=GFP-F /note="GFP, forward primer. Does NOT anneal to EGFP" terminator 2217..2288 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 2304..2331 /label=T7Te terminator /note="phage T7 early transcription terminator" primer_bind complement(2374..2391) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2508..3053) /direction=LEFT /label=p15A ori /note="Plasmids containing the medium-copy-number p15A origin of replication can be propagated in E. coli cells that contain a second plasmid with the ColE1 origin. " primer_bind complement(2588..2607) /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tcagataaaa tatttctaga tttcagtgca atttatctct tcaaatgtag cacctgaagt 61 cagccccata cgatataagt tgttactagt gcttggattc tcaccaataa aaaacgcccg 121 gcggcaaccg agcgttctga acaaatccag atggagttct gaggtcatta ctggatctat 181 caacaggagt ccaagcgagc tctcgaaccc cagagtcccg ctcagaagaa ctcgtcaaga 241 aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag 301 cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa cgctatgtcc 361 tgatagcggt ccgccacacc cagccggcca cagtcgatga atccagaaaa gcggccattt 421 tccaccatga tattcggcaa gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg 481 ggcatgcgcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg atgctcttcg 541 tccagatcat cctgatcgac aagaccggct tccatccgag tacgtgctcg ctcgatgcga 601 tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt 661 gcatcagcca tgatggatac tttctcggca ggagcaaggt gagatgacag gagatcctgc 721 cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac gtcgagcaca 781 gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc gtcctgcagt 841 tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac 901 agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc atagccgaat 961 agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc aatcatgcga 1021 aacgatcctc atcctgtctc ttgatcagat catgatcccc tgcgccatca gatccttggc 1081 ggcaagaaag ccatccagtt tactttgcag ggcttcccaa ccttaccaga gggcgcccca 1141 gctggcaatt ccgacgtcga attcaaaaga tcttaagtaa gtaagagtat acgtatatcg 1201 gctaataacg tattaaggcg cttcggcgcc tttttttatg ggggtatttt catcccaatc 1261 cacacgtcca acgcacagca aacaccacgt cgaccctatc agctgcgtgc tttctatgag 1321 tcgttgctgc ataacaaaaa gagtattgac ttaaagtcta acctataggt ataatgtgtg 1381 gagggcccaa gttcacttaa aaaggagatc aacaatgaaa gcaattttcg tactgaaaca 1441 tcttaatcat gcacaggaga ctttctaatg agcaaaggag aagaactttt cactggagtt 1501 gtcccaattc ttgttgaatt agatggtgat gttaatgggc acaaattttc tgtccgtgga 1561 gagggtgaag gtgatgctac aaacggaaaa ctcaccctta aatttatttg cactactgga 1621 aaactacctg ttccgtggcc aacacttgtc actactctga cctatggtgt tcaatgcttt 1681 tcccgttatc cggatcacat gaaacggcat gactttttca agagtgccat gcccgaaggt 1741 tatgtacagg aacgcactat atctttcaaa gatgacggga cctacaagac gcgtgctgaa 1801 gtcaagtttg aaggtgatac ccttgttaat cgtatcgagt taaagggtat tgattttaaa 1861 gaagatggaa acattcttgg acacaaactc gagtacaact ttaactcaca caatgtatac 1921 atcacggcag acaaacaaaa gaatggaatc aaagctaact tcaaaattcg ccacaacgtt 1981 gaagatggtt ccgttcaact agcagaccat tatcaacaaa atactccaat tggcgatggc 2041 cctgtccttt taccagacaa ccattacctg tcgacacaat ctgtcctttc gaaagatccc 2101 aacgaaaagc gtgaccacat ggtccttctt gagtttgtaa ctgctgctgg gattacacat 2161 ggcatggatg agctctacaa ataaggatcg gttgtcgagt aaggatctcc aggcatcaaa 2221 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga 2281 acgctctcta ctagagtcac actggctcac cttcgggtgg gcctttctgc gtttatagga 2341 tcctaactcg agcctaggga tatattccgc ttcctcgctc actgactcgc tacgctcggt 2401 cgttcgactg cggcgagcgg aaatggctta cgaacggggc ggagatttcc tggaagatgc 2461 caggaagata cttaacaggg aagtgagagg gccgcggcaa agccgttttt ccataggctc 2521 cgcccccctg acaagcatca cgaaatctga cgctcaaatc agtggtggcg aaacccgaca 2581 ggactataaa gataccaggc gtttccccct ggcggctccc tcgtgcgctc tcctgttcct 2641 gcctttcggt ttaccggtgt cattccgctg ttatggccgc gtttgtctca ttccacgcct 2701 gacactcagt tccgggtagg cagttcgctc caagctggac tgtatgcacg aaccccccgt 2761 tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggaaagaca 2821 tgcaaaagca ccactggcag cagccactgg taattgattt agaggagtta gtcttgaagt 2881 catgcgccgg ttaaggctaa actgaaagga caagttttgg tgactgcgct cctccaagcc 2941 agttacctcg gttcaaagag ttggtagctc agagaacctt cgaaaaaccg ccctgcaagg 3001 cggttttttc gttttcagag caagagatta cgcgcagacc aaaacgatct caagaagatc 3061 atcttattaa //
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