质粒名称: | pBADZ-HisCre |
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出品公司: | addgene |
目录编号: | 111187 |
存储实验室: | Tim Richmond |
载体骨架基本信息 |
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载体名称: | pBAD22 |
空载体大小: | 6427 bp |
完整质粒大小: | 6425bp |
载体类型: | 细菌细胞表达 |
载体抗性: | Bleocin (Zeocin) |
生长温度: | 37 ℃ |
宿主菌: | DH5a |
拷贝数: | 高拷贝 |
插入基因信息 |
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插入基因名称: | Cre recombinase |
标签/融合蛋白: | His tag (N terminal on insert) |
文献引用方法 |
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材料和方法部分: | pBADZ-HisCre was a gift from Tim Richmond (Addgene plasmid # 111187 ; http://n2t.net/addgene:111187 ; RRID:Addgene_111187) |
参考文献部分: | Baculovirus expression system for heterologous multiprotein complexes. Berger I, Fitzgerald DJ, Richmond TJ. Nat Biotechnol. 2004 Dec;22(12):1583-7. doi: 10.1038/nbt1036. Epub 2004 Nov 28. 10.1038/nbt1036 PubMed 15568020 |
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载体序列
LOCUS pBADZ-HisCre 6425 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Helper plasmid containing Cre-recombinase gene, which is expressed in DH10MultiBac cells. ACCESSION . VERSION . KEYWORDS pBADZ-HisCre SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6425) AUTHORS Berger I, Fitzgerald DJ, Richmond TJ TITLE Baculovirus expression system for heterologous multiprotein complexes. JOURNAL Nat Biotechnol. 2004 Dec;22(12):1583-7. doi: 10.1038/nbt1036. Epub 2004 Nov 28. PUBMED 15568020 REFERENCE 2 (bases 1 to 6425) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..6425 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 63..82 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" CDS complement(262..1140) /codon_start=1 /gene="araC" /product="L-arabinose regulatory protein" /label=araC /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE EKVNDVAVKLS" promoter 1167..1451 /gene="araBAD" /label=araBAD promoter /note="promoter of the L-arabinose operon of E. coli; the araC regulatory gene is transcribed in the opposite direction (Guzman et al., 1995)" primer_bind 1374..1393 /label=pBAD Forward /note="For vectors with E. coli araBAD promoter, forward primer" CDS 1492..1509 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 1510..2541 /codon_start=1 /product="site-specific recombinase" /label=Cre /note="Cre is a site-specific recombinase from bacteriophage P1. Recombination occurs at loxP sequences." /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE DGD" primer_bind complement(2669..2686) /label=pBAD Reverse /note="For vectors with E. coli araBAD promoter, reverse primer" terminator 2839..2925 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 3017..3044 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" promoter 3063..3154 /gene="bla" /label=AmpR promoter primer_bind complement(3373..3392) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" CDS complement(3525..3899) /codon_start=1 /gene="Sh ble from Streptoalloteichus hindustanus" /product="antibiotic-binding protein" /label=BleoR /note="confers resistance to bleomycin, phleomycin, and Zeocin(TM)" /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR EFALRDPAGNCVHFVAEEQD" promoter complement(3918..3965) /label=EM7 promoter /note="synthetic bacterial promoter " promoter complement(3972..4379) /gene="S. cerevisiae TEF1" /label=TEF1 promoter /note="promoter for EF-1-alpha" rep_origin 4837..5292 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4924..4943) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5134..5155 /label=F1ori-F /note="F1 origin, forward primer" rep_origin 5403..5991 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5892..5911 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6145..6162 /label=L4440 /note="L4440 vector, forward primer" primer_bind 6328..6350 /label=pGEX 3' /note="pGEX vectors, reverse primer" ORIGIN 1 accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca 61 tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc 121 acgatgcgtc cggcgtagag gatctgctca tgtttgacag cttatcatcg atgcataatg 181 tgcctgtcaa atggacgaag cagggattct gcaaacccta tgctactccg tcaagccgtc 241 aattgtctga ttcgttacca attatgacaa cttgacggct acatcattca ctttttcttc 301 acaaccggca cggaactcgc tcgggctggc cccggtgcat tttttaaata cccgcgagaa 361 atagagttga tcgtcaaaac caacattgcg accgacggtg gcgataggca tccgggtggt 421 gctcaaaagc agcttcgcct ggctgatacg ttggtcctcg cgccagctta agacgctaat 481 ccctaactgc tggcggaaaa gatgtgacag acgcgacggc gacaagcaaa catgctgtgc 541 gacgctggcg atatcaaaat tgctgtctgc caggtgatcg ctgatgtact gacaagcctc 601 gcgtacccga ttatccatcg gtggatggag cgactcgtta atcgcttcca tgcgccgcag 661 taacaattgc tcaagcagat ttatcgccag cagctccgaa tagcgccctt ccccttgccc 721 ggcgttaatg atttgcccaa acaggtcgct gaaatgcggc tggtgcgctt catccgggcg 781 aaagaacccc gtattggcaa atattgacgg ccagttaagc cattcatgcc agtaggcgcg 841 cggacgaaag taaacccact ggtgatacca ttcgcgagcc tccggatgac gaccgtagtg 901 atgaatctct cctggcggga acagcaaaat atcacccggt cggcaaacaa attctcgtcc 961 ctgatttttc accaccccct gaccgcgaat ggtgagattg agaatataac ctttcattcc 1021 cagcggtcgg tcgataaaaa aatcgagata accgttggcc tcaatcggcg ttaaacccgc 1081 caccagatgg gcattaaacg agtatcccgg cagcagggga tcattttgcg cttcagccat 1141 acttttcata ctcccgccat tcagagaaga aaccaattgt ccatattgca tcagacattg 1201 ccgtcactgc gtcttttact ggctcttctc gctaaccaaa ccggtaaccc cgcttattaa 1261 aagcattctg taacaaagcg ggaccaaagc catgacaaaa acgcgtaaca aaagtgtcta 1321 taatcacggc agaaaagtcc acattgatta tttgcacggc gtcacacttt gctatgccat 1381 agcattttta tccataagat tagcggatcc tacctgacgc tttttatcgc aactctctac 1441 tgtttctcca tacccgtttt tttgggctag caggaggaat tcaccatggc acatcatcat 1501 caccaccaca tgtccaattt actgaccgta caccaaaatt tgcctgcatt accggtcgat 1561 gcaacgagtg atgaggttcg caagaacctg atggacatgt tcagggatcg ccaggcgttt 1621 tctgagcata cctggaaaat gcttctgtcc gtttgccggt cgtgggcggc atggtgcaag 1681 ttgaataacc ggaaatggtt tcccgcagaa cctgaagatg ttcgcgatta tcttctatat 1741 cttcaggcgc gcggtctggc agtaaaaact atccagcaac atttgggcca gctaaacatg 1801 cttcatcgtc ggtccgggct gccacgacca agtgacagca atgctgtttc actggttatg 1861 cggcggatcc gaaaagaaaa cgttgatgcc ggtgaacgtg caaaacaggc tctagcgttc 1921 gaacgcactg atttcgacca ggttcgttca ctcatggaaa atagcgatcg ctgccaggat 1981 atacgtaatc tggcatttct ggggattgct tataacaccc tgttacgtat agccgaaatt 2041 gccaggatca gggttaaaga tatctcacgt actgacggtg ggagaatgtt aatccatatt 2101 ggcagaacga aaacgctggt tagcaccgca ggtgtagaga aggcacttag cctgggggta 2161 actaaactgg tcgagcgatg gatttccgtc tctggtgtag ctgatgatcc gaataactac 2221 ctgttttgcc gggtcagaaa aaatggtgtt gccgcgccat ctgccaccag ccagctatca 2281 actcgcgccc tggaagggat ttttgaagca actcatcgat tgatttacgg cgctaaggat 2341 gactctggtc agagatacct ggcctggtct ggacacagtg cccgtgtcgg agccgcgcga 2401 gatatggccc gcgctggagt ttcaataccg gagatcatgc aagctggtgg ctggaccaat 2461 gtaaatattg tcatgaacta tatccgtaac ctggatagtg aaacaggggc aatggtgcgc 2521 ctgctggaag atggcgatta gccattaacg cgtaaatgat tgctataatt agttgatatt 2581 atctagagct gggtacccgg ggatcctcta gagtcgacct gcaggcatgc aagcttggct 2641 gttttggcgg atgagagaag attttcagcc tgatacagat taaatcagaa cgcagaagcg 2701 gtctgataaa acagaatttg cctggcggca gtagcgcggt ggtcccacct gaccccatgc 2761 cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc catgcgagag 2821 tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt 2881 tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gggagcggat 2941 ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc ataaactgcc 3001 aggcatcaaa ttaagcagaa ggccatcctg acggatggcc tttttgcgtt tctacaaact 3061 cttttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg 3121 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc 3181 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt 3241 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct 3301 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac 3361 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact 3421 cggtcgccgc atacactatt ctcagaatga cttggttgag atcgacaaag gaaaaggggg 3481 acggatctcc gaggcctcgg acccgtcggg ccgcgtcgga cgtgtcagtc ctgctcctcg 3541 gccacgaagt gcacgcagtt gccggccggg tcgcgcaggg cgaactcccg cccccacggc 3601 tgctcgccga tctcggtcat ggccggcccg gaggcgtccc ggaagttcgt ggacacgacc 3661 tccgaccact cggcgtacag ctcgtccagg ccgcgcaccc acacccaggc cagggtgttg 3721 tccggcacca cctggtcctg gaccgcgctg atgaacaggg tcacgtcgtc ccggaccaca 3781 ccggcgaagt cgtcctccac gaagtcccgg gagaacccga gccggtcggt ccagaactcg 3841 accgctccgg cgacgtcgcg cgcggtgagc accggaacgg cactggtcaa cttggccatg 3901 gtttagttcc tcaccttgtc gtattatact atgccgatat actatgccga tgattaattg 3961 tcaacaccgc ccttagatta gattgctatg ctttctttct aatgagcaag aagtaaaaaa 4021 agttgtaata gaacaagaaa aatgaaactg aaacttgaga aattgaagac cgtttattaa 4081 cttaaatatc aatgggaggt catcgaaaga gaaaaaaatc aaaaaaaaaa attttcaaga 4141 aaaagaaacg tgataaaaat ttttattgcc tttttcgacg aagaaaaaga aacgaggcgg 4201 tctctttttt cttttccaaa cctttagtac gggtaattaa cgacacccta gaggaagaaa 4261 gaggggaaat ttagtatgct gtgcttgggt gttttgaagt ggtacggcga tgcgcggagt 4321 ccgagaaaat ctggaagagt aaaaaaggag tagaaacatt ttgaagctat ggtgtgtggg 4381 ggatccgcac aaacgaaggt ctcacttaat cttctgtact ctgaagagga gtgggaaata 4441 ccaagaaaaa catcaaactc gaatgatttt cccaaacccc taccacaaga tattcatcag 4501 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat 4561 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat 4621 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc 4681 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga 4741 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc 4801 agaccaagtt tactcatata tactttagat tgatttacgc gccctgtagc ggcgcattaa 4861 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc 4921 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag 4981 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca 5041 aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc 5101 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa 5161 cactcaaccc tatctcgggc tattcttttg atttataagg gattttgccg atttcggcct 5221 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa 5281 cgtttacaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc 5341 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct 5401 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta 5461 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc 5521 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac 5581 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct 5641 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat 5701 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg 5761 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa 5821 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg 5881 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga 5941 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc 6001 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct 6061 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct 6121 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg 6181 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatagg gtcatggctg 6241 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat 6301 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt 6361 catcaccgaa acgcgcgagg cagaaggaga tggcgcccaa cagtcccccg gccacggggc 6421 ctgcc //
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