纽普生物

pET-30a(+)载体

载体基本信息
载体名称 pET-30a(+) 载体抗性 Kanamycin
载体长度 5422 bp 载体类型 pET & Duet Vectors (Novagen)
载体来源 Novagen (EMD Millipore) 拷贝数 High copy number

pET-30a(+)载体质粒图谱

pET-30a(+)载体质粒图谱

pET-30a(+)载体载体序列

LOCUS       Exported                5422 bp ds-DNA    circular SYN 02-12-2013
DEFINITION  Bacterial vector for expression of N-terminally S-tagged proteins 
            with an enterokinase site. For other reading frames, use pET-30b(+) 
            or pET-30c(+).
ACCESSION   .
VERSION     .
KEYWORDS    pET-30a(+)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5422)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
  JOURNAL   Exported 2017/2/28 from SnapGene 2.3.2
            http://www.snapgene.com
COMMENT     
FEATURES             Location/Qualifiers
     source          1..5422
                     /organism="synthetic DNA construct"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
     terminator      26..73
                     /note="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     misc_feature    158..217
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             complement(219..233)
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /note="enterokinase site"
                     /translation="DDDDK"
     CDS             complement(249..293)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /note="S-Tag"
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(300..317)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /note="thrombin site"
                     /translation="LVPRGS"
     CDS             complement(327..344)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     CDS             complement(345..347)
                     /codon_start=1
                     /product="start codon"
                     /note="ATG"
                     /translation="M"
     RBS             356..361
                     /note="ribosome binding site"
     protein_bind    392..416
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /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)."
     promoter        complement(417..435)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        748..825
                     /gene="lacI"
                     /note="lacI promoter"
                     /note="
                     "
     CDS             826..1908
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /note="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"
     CDS             2717..2908
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein"
                     /note="rop"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     rep_origin      complement(3338..3926)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             4048..4863
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /note="KanR"
                     /note="confers resistance to kanamycin"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      complement(4956..5411)
                     /direction=LEFT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt
      181 cgacggagct cgaattcgga tccgatatca gccatggcct tgtcgtcgtc gtcggtaccc
      241 agatctgggc tgtccatgtg ctggcgttcg aatttagcag cagcggtttc tttcatacca
      301 gaaccgcgtg gcaccagacc agaagaatga tgatgatgat ggtgcatatg tatatctcct
      361 tcttaaagtt aaacaaaatt atttctagag gggaattgtt atccgctcac aattccccta
      421 tagtgagtcg tattaatttc gcgggatcga gatcgatctc gatcctctac gccggacgca
      481 tcgtggccgg catcaccggc gccacaggtg cggttgctgg cgcctatatc gccgacatca
      541 ccgatgggga agatcgggct cgccacttcg ggctcatgag cgcttgtttc ggcgtgggta
      601 tggtggcagg ccccgtggcc gggggactgt tgggcgccat ctccttgcat gcaccattcc
      661 ttgcggcggc ggtgctcaac ggcctcaacc tactactggg ctgcttccta atgcaggagt
      721 cgcataaggg agagcgtcga gatcccggac accatcgaat ggcgcaaaac ctttcgcggt
      781 atggcatgat agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg
      841 ttatacgatg tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac
      901 caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg
      961 aattacattc ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt gctgattggc
     1021 gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct
     1081 cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa
     1141 gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg tcagtgggct gatcattaac
     1201 tatccgctgg atgaccagga tgccattgct gtggaagctg cctgcactaa tgttccggcg
     1261 ttatttcttg atgtctctga ccagacaccc atcaacagta ttattttctc ccatgaagac
     1321 ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc accagcaaat cgcgctgtta
     1381 gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc
     1441 actcgcaatc aaattcagcc gatagcggaa cgggaaggcg actggagtgc catgtccggt
     1501 tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat gctggttgcc
     1561 aacgatcaga tggcgctggg cgcaatgcgc gccattaccg agtccgggct gcgcgttggt
     1621 gcggacatct cggtagtggg atacgacgat accgaagaca gctcatgtta tatcccgccg
     1681 ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg
     1741 caactctctc agggccaggc ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa
     1801 agaaaaacca ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     1861 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     1921 taatgtaagt tagctcactc attaggcacc gggatctcga ccgatgccct tgagagcctt
     1981 caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac
     2041 tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctctggg tcattttcgg
     2101 cgaggaccgc tttcgctgga gcgcgacgat gatcggcctg tcgcttgcgg tattcggaat
     2161 cttgcacgcc ctcgctcaag ccttcgtcac tggtcccgcc accaaacgtt tcggcgagaa
     2221 gcaggccatt atcgccggca tggcggcccc acgggtgcgc atgatcgtgc tcctgtcgtt
     2281 gaggacccgg ctaggctggc ggggttgcct tactggttag cagaatgaat caccgatacg
     2341 cgagcgaacg tgaagcgact gctgctgcaa aacgtctgcg acctgagcaa caacatgaat
     2401 ggtcttcggt ttccgtgttt cgtaaagtct ggaaacgcgg aagtcagcgc cctgcaccat
     2461 tatgttccgg atctgcatcg caggatgctg ctggctaccc tgtggaacac ctacatctgt
     2521 attaacgaag cgctggcatt gaccctgagt gatttttctc tggtcccgcc gcatccatac
     2581 cgccagttgt ttaccctcac aacgttccag taaccgggca tgttcatcat cagtaacccg
     2641 tatcgtgagc atcctctctc gtttcatcgg tatcattacc cccatgaaca gaaatccccc
     2701 ttacacggag gcatcagtga ccaaacagga aaaaaccgcc cttaacatgg cccgctttat
     2761 cagaagccag acattaacgc ttctggagaa actcaacgag ctggacgcgg atgaacaggc
     2821 agacatctgt gaatcgcttc acgaccacgc tgatgagctt taccgcagct gcctcgcgcg
     2881 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     2941 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     3001 gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata ctggcttaac
     3061 tatgcggcat cagagcagat tgtactgaga gtgcaccata tatgcggtgt gaaataccgc
     3121 acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact
     3181 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     3241 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     3301 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     3361 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     3421 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3481 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     3541 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3601 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     3661 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     3721 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     3781 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     3841 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     3901 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     3961 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gaacaataaa actgtctgct
     4021 tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac gtcttgctct
     4081 aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat
     4141 aatgtcgggc aatcaggtgc gacaatctat cgattgtatg ggaagcccga tgcgccagag
     4201 ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga
     4261 ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct
     4321 gatgatgcat ggttactcac cactgcgatc cccgggaaaa cagcattcca ggtattagaa
     4381 gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg
     4441 cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag
     4501 gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat
     4561 ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat
     4621 tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta
     4681 ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc
     4741 ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat
     4801 ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc
     4861 taagaattaa ttcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     4921 ggttccgcgc acatttcccc gaaaagtgcc acctgaaatt gtaaacgtta atattttgtt
     4981 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     5041 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     5101 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     5161 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     5221 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     5281 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     5341 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     5401 acagggcgcg tcccattcgc ca
//

pET-30a(+)载体载体特征

一级属性 二级属性 属性值 起始位置
常规长度5422
常规描述Bacterial vector for expression of N-terminally S-tagged proteins with an enterokinase site. For other reading frames, use pET-30b(+) or pET-30c(+).
常规是否环形1
常规作者Novagen (EMD Millipore)
常规注释
来源实验宿主Escherichia coli1..5422
来源分子类型other DNA1..5422
来源种属synthetic DNA construct1..5422
终止子备注T7 terminator,transcription terminator for bacteriophage T7 RNA polymerase26..73
编码区翻译起始位点1140..157
编码区备注6xHis140..157
编码区产物6xHis affinity tag140..157
编码区翻译序列HHHHHH140..157
其它特征区备注MCS,multiple cloning site158..217
编码区翻译起始位点1219..233
编码区备注enterokinase site219..233
编码区产物enterokinase recognition and cleavage site219..233
编码区翻译序列DDDDK219..233
编码区翻译起始位点1249..293
编码区备注S-Tag249..293
编码区产物affinity and epitope tag derived from pancreatic ribonuclease A249..293
编码区翻译序列KETAAAKFERQHMDS249..293
编码区翻译起始位点1300..317
编码区备注thrombin site300..317
编码区产物thrombin recognition and cleavage site300..317
编码区翻译序列LVPRGS300..317
编码区翻译起始位点1327..344
编码区备注6xHis327..344
编码区产物6xHis affinity tag327..344
编码区翻译序列HHHHHH327..344
编码区翻译起始位点1345..347
编码区备注ATG345..347
编码区产物start codon345..347
编码区翻译序列M345..347
核糖体结合位点备注ribosome binding site356..361
蛋白结合位点嵌合范围lac repressor encoded by lacI392..416
蛋白结合位点备注lac operator,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).392..416
启动子备注T7 promoter,promoter for bacteriophage T7 RNA polymerase417..435
启动子基因lacI748..825
启动子备注lacI promoter, 748..825
编码区翻译起始位点1826..1908
编码区基因lacI826..1908
编码区备注lacI,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).826..1908
编码区产物lac repressor826..1908
编码区翻译序列MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ826..1908
编码区翻译起始位点12717..2908
编码区基因rop2717..2908
编码区备注rop2717..2908
编码区产物Rop protein2717..2908
编码区翻译序列MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHADELYRSCLARFGDDGENL2717..2908
复制起点方向LEFT3338..3926
复制起点备注ori,high-copy-number ColE1/pMB1/pBR322/pUC origin of replication3338..3926
编码区翻译起始位点14048..4863
编码区基因aph(3')-Ia4048..4863
编码区备注KanR,confers resistance to kanamycin4048..4863
编码区产物aminoglycoside phosphotransferase4048..4863
编码区翻译序列MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF4048..4863
复制起点方向LEFT4956..5411
复制起点备注f1 ori,f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis4956..5411

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