pMZ0002

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联系方式:I47-825O-882O

相关技术服务:质粒构建    基因合成    质粒大提

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载体基本信息

质粒名称: pMZ0002
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
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
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
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

载体图谱质粒图谱和多克隆位点信息

pMZ0002载体图谱

载体简介

载体序列

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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