质粒名称: | pMZ0002 |
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出品公司: | addgene |
目录编号: | 102239 |
存储实验室: | Axel Brunger |
载体骨架基本信息 |
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载体名称: | pPROEX1 |
完整质粒大小: | 6914 bp |
载体类型: | 细菌细胞表达 |
载体抗性: | 氨苄霉素 |
生长温度: | 30℃ |
宿主菌: | DH5a |
拷贝数: | 未知 |
插入基因信息 |
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插入基因名称: | nameN-ethylmaleimide sensitive factor/NSF |
物种来源: | Chinese hamster |
标签/融合蛋白: | 6x His tag (N terminal on backbone) |
文献引用方法 |
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材料和方法部分: | pMZ0002 was a gift from Axel Brunger (Addgene plasmid # 102239 ; http://n2t.net/addgene:102239 ; RRID:Addgene_102239) |
参考文献部分: | Mechanistic insights into the recycling machine of the SNARE complex. Zhao M, Wu S, Zhou Q, Vivona S, Cipriano DJ, Cheng Y, Brunger AT. Nature. 2015 Feb 5;518(7537):61-7. doi: 10.1038/nature14148. Epub 2015 Jan 12. 10.1038/nature14148 PubMed 25581794 |
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LOCUS pMZ0002 6914 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses Chinese hamster NSF in bacteria. ACCESSION . VERSION . KEYWORDS pMZ0002 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6914) AUTHORS Zhao M, Wu S, Zhou Q, Vivona S, Cipriano DJ, Cheng Y, Brunger AT TITLE Mechanistic insights into the recycling machine of the SNARE complex. JOURNAL Nature. 2015 Feb 5;518(7537):61-7. doi: 10.1038/nature14148. Epub 2015 Jan 12. PUBMED 25581794 REFERENCE 2 (bases 1 to 6914) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..6914 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 11..599 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 500..519 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 753..770 /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(847..1275) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(957..978) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 1169..1188 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind complement(1313..1332) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 1473..1495 /label=pGEX 3' /note="pGEX vectors, reverse primer" promoter 1573..1650 /gene="lacI (mutant)" /label=lacIq promoter /note="In the lacIq allele, a single base change in the promoter boosts expression of the lacI gene about 10-fold." CDS 1651..2733 /codon_start=1 /gene="lacI" /product="lac repressor" /label=lacI /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" primer_bind complement(1670..1689) /label=LacI-R /note="LacI, reverse primer" protein_bind 3002..3018 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." primer_bind 3007..3029 /label=M13/pUC Reverse /note="In lacZ gene" CDS 3045..3062 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 3084..3104 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" primer_bind complement(5408..5425) /label=pBAD Reverse /note="For vectors with E. coli araBAD promoter, reverse primer" terminator 5578..5664 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 5756..5783 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" promoter 5802..5893 /gene="bla" /label=AmpR promoter CDS 5894..6754 /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 complement(6112..6131) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" ORIGIN 1 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac 61 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg 121 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag 181 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac 241 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt 301 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg 361 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc 421 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc 481 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc 541 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa 601 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt 661 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg 721 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag 781 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcataattt 841 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa 901 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag 961 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg 1021 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga 1081 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg 1141 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg 1201 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc 1261 cgctacaggg cgcgtcccat tcgccattca ggctgctatg gtgcactctc agtacaatct 1321 gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat 1381 ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc 1441 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc 1501 accgtcatca ccgaaacgcg cgaggcagca gatcaattcg cgcgcgaagg cgaagcggca 1561 tgcatttacg ttgacaccat cgaatggtgc aaaacctttc gcggtatggc atgatagcgc 1621 ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca 1681 gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt 1741 tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac 1801 cgcgtggcac aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt 1861 ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg 1921 ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg 1981 gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac 2041 caggatgcca ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc 2101 tctgaccaga cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc 2161 gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt 2221 tctgtctcgg cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt 2281 cagccgatag cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg 2341 caaatgctga atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg 2401 ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatctcggta 2461 gtgggatacg acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa 2521 caggattttc gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc 2581 caggcggtga agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg 2641 gcacccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca 2701 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg taagttagcg 2761 cgaattgatc tggtttgaca gcttatcatc gactgcacgg tgcaccaatg cttctggcgt 2821 caggcagcca tcggaagctg tggtatggct gtgcaggtcg taaatcactg cataattcgt 2881 gtcgctcaag gcgcactccc gttctggata atgttttttg cgccgacatc ataacggttc 2941 tggcaaatat tctgaaatga gctgttgaca attaatcatc cggtccgtat aatctgtgga 3001 attgtgagcg gataacaatt tcacacagga aacagaccat gggtcatcat catcatcatc 3061 acgattacga tatcccaacg accgaaaacc tgtattttca gggcgcccat atggcgggcc 3121 ggagtatgca agctgcgagg tgccctaccg atgaattgtc cttaagcaac tgtgcagttg 3181 tgagcgagaa ggattaccag tctggacagc atgtgattgt gaggacctcc cccaatcaca 3241 aatacatatt tacactgagg acccatccgt cggtggttcc cggaagcgtt gcattcagct 3301 tacctcagcg aaaatgggct ggactttcta ttgggcagga aatagaagtc gccttgtatt 3361 catttgacaa agccaagcaa tgcatcggca ccatgaccat cgagattgac tttctgcaga 3421 aaaaaaacat cgactccaac ccatatgata ccgacaagat ggcagcagaa ttcatccagc 3481 agttcaacaa ccaggccttc tcagtgggac agcagcttgt ctttagcttt aatgataagc 3541 tcttcggatt actggtgaag gacattgaag caatggaccc tagcatcctg aagggagagc 3601 ctgcatcagg caaaaggcag aagattgaag tcggactggt tgttggaaat agtcaagttg 3661 catttgaaaa agcagaaaat tcatcgctca atcttattgg caaagctaaa accaaggaaa 3721 atcgccagag tattatcaat cctgactgga actttgagaa aatgggaatc ggcggtctgg 3781 ataaggagtt ttcagatatt ttccgacgag catttgcttc tcgggtgttc cctccggaga 3841 ttgtggagca gatgggttgt aaacatgtta aaggcatcct gttgtacggc ccccctggtt 3901 gtggtaagac tctcttggct cgacagattg gcaagatgct gaatgcgaga gaacccaagg 3961 tggtcaatgg gccagaaatc cttaacaaat atgtgggaga atcggaggcg aacattcgta 4021 aactttttgc tgatgccgaa gaggagcaaa ggaggcttgg tgctaacagt ggcttacaca 4081 tcatcatctt tgatgaaatc gatgccatct gcaagcagag agggagcatg gctggcagca 4141 ccggagttca cgacaccgtt gtcaaccagc tgctgtccaa gattgatggc gttgagcagc 4201 tgaacaacat cctggttatt gggatgacca atagaccaga tttgatagat gaggctctcc 4261 ttcgacctgg aagactggaa gttaaaatgg agataggctt gccagatgag aagggtcgac 4321 tgcagatcct tcatatccac acagcaagga tgagaggaca tcagttactg tctgcagatg 4381 tggacattaa ggaactggcc gtggagacta agaatttcag tggcgctgag ctggaaggtc 4441 tggtgcgagc agcacagtcg acggccatga accgacacat aaaggccagt acgaaagtgg 4501 aagtagacat ggagaaagca gagagtctgc aggtgacaag aggagacttc ctggcttctt 4561 tggagaatga tatcaaacca gcttttggca caaaccaaga ggattatgcc agttacatta 4621 tgaatggaat catcaagtgg ggtgaccctg tgacccgtgt cctggatgac ggagagctgc 4681 tggtccagca gactaaaaac agtgaccgta cgcctctggt tagcgtcctt ctggaagggc 4741 cccctcacag cgggaagact gccctagctg caaagatcgc agaggaatcc aactttccct 4801 tcatcaagat ctgctccccc gataagatga ttggcttttc tgagacagcc aagtgtcagg 4861 ccatgaagaa gatctttgat gatgcttaca aatctcaact cagttgtgtg gtggtggatg 4921 acatcgaaag gctacttgat tatgtcccca ttggcccccg gttctccaac ctggtgttgc 4981 aagctcttct tgtgttacta aagaaggcac ctcctcaggg ccgcaagctt ctcatcatcg 5041 ggaccaccag ccgcaaagat gtccttcagg aaatggagat gctgaatgcc ttcagcacca 5101 ccatccacgt gcccaacatt gccacaggag agcagctgct ggaagctttg gagcttctgg 5161 gcaacttcaa agataaggaa cgcaccacaa ttgctcagca agtcaaaggg aagaaggtct 5221 ggataggaat caagaagtta ctaatgttga ttgagatgtc cctgcagatg gaccctgagt 5281 atcgggtgag gaaattcttg gccctcttga gagaagaagg cgctagcccc ctggactttg 5341 attgaggatc cctcgaggca tgcggtacca agcttggctg ttttggcgga tgagagaaga 5401 ttttcagcct gatacagatt aaatcagaac gcagaagcgg tctgataaaa cagaatttgc 5461 ctggcggcag tagcgcggtg gtcccacctg accccatgcc gaactcagaa gtgaaacgcc 5521 gtagcgccga tggtagtgtg gggtctcccc atgcgagagt agggaactgc caggcatcaa 5581 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg 5641 aacgctctcc tgagtaggac aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg 5701 cccggagggt ggcgggcagg acgcccgcca taaactgcca ggcatcaaat taagcagaag 5761 gccatcctga cggatggcct ttttgcgttt ctacaaactc ttttgtttat ttttctaaat 5821 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg 5881 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc 5941 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga 6001 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga 6061 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg 6121 cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc 6181 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac 6241 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact 6301 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca 6361 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg 6421 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact 6481 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg 6541 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg 6601 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat 6661 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc 6721 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat 6781 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt 6841 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc 6901 cgtagaaaag atca //
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