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pET-30c(+)

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  • YB-0035
  • 2025年07月12日
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    • 详细信息
    • 文献和实验
    • 技术资料
    • 保存条件：

      -20℃

    • 保质期：

      6个月

    • 英文名：

      pET-30c(+)

    • 库存：

      大量

    • 供应商：

      钰博生物

    • 规格：

      1ug/5ug

    pET-30c(+)

    pET30c载体基本信息

    出品公司: Ybscience
    别名: pET30c, pet 30c
    质粒类型: 大肠杆菌蛋白表达
    表达水平: 高
    克隆方法: 多克隆位点，限制性内切酶
    载体大小: 5423 bp
    5' 测序引物: T7
    5' 测序引物序列:   5'-TAATACGACTCACTATAGGG-3'
    载体标签: N-His, N-S, N-Thrombin, N-EK, C-His
    载体抗性: Kanamycin
    表达宿主菌: BL21(DE3) 、 Rosetta2(DE3)、C43(DE3)
    备注: N term thrombin cleavage site; N term enterokinase cleavage site; a,b,c vary by MCS
    稳定性: 瞬时表达 Transient
    组成型: 组成型 Constitutive
    病毒/非病毒: 非病毒

    pET30c载体质粒图谱和多克隆位点信息

    pet30c载体图谱
    pet30c多克隆位点

    Feature Name Start End  
    T7_terminator 129 1
    T7_Terminal_primer 69 87
    EK 231 220
    lacI 303 1909
    T7_transl_en_RBS 373 357
    lacO 418 391
    T7_promoter 436 418
    tet (300 - 563) 472 735
    pBRrevBam_primer 543 524
    tet (576 - 851) 1968 2243
    ROP 2718 2909
    pGEX_3_primer 2925 2903
    pBR322_origin 3943 3324
    KanR2 4049 4864
    ORF Start End  
    ORF frame 1 328 1077
    ORF frame 2 950 1909
    ORF frame 2 4049 4864
    Enzyme Name Cut
    XhoI 158
    NotI 166
    EagI 166
    HindIII 173
    SalI 179
    SacI 190
    EcoRI 192
    BamHI 198
    EcoRV 209
    NcoI 213
    KpnI 239
    BglII 242
    BstBI 269
    NdeI 347
    XbaI 385
    BclI 1191
    ApaI 1388
    HpaI 1683
    FspI 2259
    NruI 4137
    SmaI 4354
    XmaI 4352

     

    pET30c载体简介

    The pET-30a-c(+) vectors carry an N-terminal His• Tag®/ thrombin/ S• Tag™/ enterokinase configuration plus an optional C-terminal His• Tag sequence. Unique sites are shown on the circle map. Note that the sequence is numbered by the pBR322 convention, so the T7 expression region is reversed on the circular map. The cloning/expression region of the coding strand transcribed by T7 RNA polymerase is shown below. The f1 origin is oriented so that infection with helper phage will produce virions containing single-stranded DNA that corresponds to the coding strand. Therefore, single-stranded sequencing should be performed using the T7 terminator primer (Cat. No. 69337-3).

    pET30c载体序列

    LOCUS       pET-30 c (+)    5423 bp     DNA Circular    SYN
    DEFINITION  pET-30 c (+)
    ACCESSION   
    KEYWORDS    
    SOURCE      
      ORGANISM  other sequences; artificial sequences; vectors.
    COMMENT     This file is created by Vector NTI
                http://www.biofeng.com/
    COMMENT     ORIGDB|GenBank
    COMMENT     VNTAUTHORNAME|biofeng.com|
    COMMENT     VNTOAUTHORAD1|Novagen Inc.|
    FEATURES             Location/Qualifiers
         source          1..5423
                         /organism="pET-30 c (+)"
                         /mol_type="other DNA"
         terminator      complement(1..129)
                         /label="T7_terminator"
         misc_feature    69..87
                         /label="T7_Terminal_primer"
         misc_feature    complement(220..231)
                         /label="EK"
         misc_feature    303..1909
                         /label="lacI"
         CDS             328..1077
                         /label="ORF frame 1"
                         /translation="MMMMMVHMYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSI
                         SILYAGRIVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLL
                         GAISLHAPFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRE
                         SIQGGECETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPR
                         GTTTGGQTVVADWRCHLQSGPARAVANCRGD*"
         misc_feature    complement(357..373)
                         /label="T7_transl_en_RBS"
                         /translation="MMMMMVHMYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSI
                         SILYAGRIVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLL
                         GAISLHAPFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRE
                         SIQGGECETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPR
                         GTTTGGQTVVADWRCHLQSGPARAVANCRGD*"
         misc_feature    complement(391..418)
                         /label="lacO"
                         /translation="MMMMMVHMYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSI
                         SILYAGRIVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLL
                         GAISLHAPFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRE
                         SIQGGECETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPR
                         GTTTGGQTVVADWRCHLQSGPARAVANCRGD*"
         promoter        complement(418..436)
                         /label="T7_promoter"
                         /translation="MMMMMVHMYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSI
                         SILYAGRIVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLL
                         GAISLHAPFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRE
                         SIQGGECETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPR
                         GTTTGGQTVVADWRCHLQSGPARAVANCRGD*"
         gene            472..735
                         /label="tet (300 - 563)"
                         /gene="tet (300 - 563)"
                         /translation="MMMMMVHMYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSI
                         SILYAGRIVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLL
                         GAISLHAPFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRE
                         SIQGGECETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPR
                         GTTTGGQTVVADWRCHLQSGPARAVANCRGD*"
         misc_feature    complement(524..543)
                         /label="pBRrevBam_primer"
                         /translation="MMMMMVHMYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSI
                         SILYAGRIVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLL
                         GAISLHAPFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRE
                         SIQGGECETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPR
                         GTTTGGQTVVADWRCHLQSGPARAVANCRGD*"
         CDS             950..1909
                         /label="ORF frame 2"
                         /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                         RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                         PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                         HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                         SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                         LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
         gene            1968..2243
                         /label="tet (576 - 851)"
                         /gene="tet (576 - 851)"
                         /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                         RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                         PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                         HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                         SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                         LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
         misc_feature    2718..2909
                         /label="ROP"
                         /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                         RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                         PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                         HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                         SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                         LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
         misc_feature    complement(2903..2925)
                         /label="pGEX_3_primer"
                         /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                         RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                         PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                         HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                         SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                         LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
         rep_origin      complement(3324..3943)
                         /label="pBR322_origin"
                         /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                         RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                         PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                         HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                         SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                         LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
         gene            4049..4864
                         /label="KanR2"
                         /gene="KanR2"
                         /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                         RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                         PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                         HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                         SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                         LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
         CDS             4049..4864
                         /label="ORF frame 2"
                         /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLY
                         GKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIP
                         GKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGL
                         VDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVG
                         RVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"
    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 tccacagata tcccatggcc ttgtcgtcgt cgtcggtacc
      241 cagatctggg ctgtccatgt gctggcgttc gaatttagca gcagcggttt ctttcatacc
      301 agaaccgcgt ggcaccagac cagaagaatg atgatgatga tggtgcatat gtatatctcc
      361 ttcttaaagt taaacaaaat tatttctaga ggggaattgt tatccgctca caattcccct
      421 atagtgagtc gtattaattt cgcgggatcg agatcgatct cgatcctcta cgccggacgc
      481 atcgtggccg gcatcaccgg cgccacaggt gcggttgctg gcgcctatat cgccgacatc
      541 accgatgggg aagatcgggc tcgccacttc gggctcatga gcgcttgttt cggcgtgggt
      601 atggtggcag gccccgtggc cgggggactg ttgggcgcca tctccttgca tgcaccattc
      661 cttgcggcgg cggtgctcaa cggcctcaac ctactactgg gctgcttcct aatgcaggag
      721 tcgcataagg gagagcgtcg agatcccgga caccatcgaa tggcgcaaaa cctttcgcgg
      781 tatggcatga tagcgcccgg aagagagtca attcagggtg gtgaatgtga aaccagtaac
      841 gttatacgat gtcgcagagt atgccggtgt ctcttatcag accgtttccc gcgtggtgaa
      901 ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct
      961 gaattacatt cccaaccgcg tggcacaaca actggcgggc aaacagtcgt tgctgattgg
     1021 cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg cgattaaatc
     1081 tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa gcggcgtcga
     1141 agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc tgatcattaa
     1201 ctatccgctg gatgaccagg atgccattgc tgtggaagct gcctgcacta atgttccggc
     1261 gttatttctt gatgtctctg accagacacc catcaacagt attattttct cccatgaaga
     1321 cggtacgcga ctgggcgtgg agcatctggt cgcattgggt caccagcaaa tcgcgctgtt
     1381 agcgggccca ttaagttctg tctcggcgcg tctgcgtctg gctggctggc ataaatatct
     1441 cactcgcaat caaattcagc cgatagcgga acgggaaggc gactggagtg ccatgtccgg
     1501 ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt cccactgcga tgctggttgc
     1561 caacgatcag atggcgctgg gcgcaatgcg cgccattacc gagtccgggc tgcgcgttgg
     1621 tgcggacatc tcggtagtgg gatacgacga taccgaagac agctcatgtt atatcccgcc
     1681 gttaaccacc atcaaacagg attttcgcct gctggggcaa accagcgtgg accgcttgct
     1741 gcaactctct cagggccagg cggtgaaggg caatcagctg ttgcccgtct cactggtgaa
     1801 aagaaaaacc accctggcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     1861 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     1921 ttaatgtaag ttagctcact cattaggcac cgggatctcg accgatgccc ttgagagcct
     1981 tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga
     2041 ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctctgg gtcattttcg
     2101 gcgaggaccg ctttcgctgg agcgcgacga tgatcggcct gtcgcttgcg gtattcggaa
     2161 tcttgcacgc cctcgctcaa gccttcgtca ctggtcccgc caccaaacgt ttcggcgaga
     2221 agcaggccat tatcgccggc atggcggccc cacgggtgcg catgatcgtg ctcctgtcgt
     2281 tgaggacccg gctaggctgg cggggttgcc ttactggtta gcagaatgaa tcaccgatac
     2341 gcgagcgaac gtgaagcgac tgctgctgca aaacgtctgc gacctgagca acaacatgaa
     2401 tggtcttcgg tttccgtgtt tcgtaaagtc tggaaacgcg gaagtcagcg ccctgcacca
     2461 ttatgttccg gatctgcatc gcaggatgct gctggctacc ctgtggaaca cctacatctg
     2521 tattaacgaa gcgctggcat tgaccctgag tgatttttct ctggtcccgc cgcatccata
     2581 ccgccagttg tttaccctca caacgttcca gtaaccgggc atgttcatca tcagtaaccc
     2641 gtatcgtgag catcctctct cgtttcatcg gtatcattac ccccatgaac agaaatcccc
     2701 cttacacgga ggcatcagtg accaaacagg aaaaaaccgc ccttaacatg gcccgcttta
     2761 tcagaagcca gacattaacg cttctggaga aactcaacga gctggacgcg gatgaacagg
     2821 cagacatctg tgaatcgctt cacgaccacg ctgatgagct ttaccgcagc tgcctcgcgc
     2881 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     2941 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     3001 ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat actggcttaa
     3061 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atatgcggtg tgaaataccg
     3121 cacagatgcg taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac
     3181 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     3241 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     3301 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3361 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     3421 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3481 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3541 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3601 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3661 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3721 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     3781 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     3841 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3901 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     3961 gctcagtgga acgaaaactc acgttaaggg attttggtca tgaacaataa aactgtctgc
     4021 ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcttgctc
     4081 taggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     4141 taatgtcggg caatcaggtg cgacaatcta tcgattgtat gggaagcccg atgcgccaga
     4201 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     4261 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     4321 tgatgatgca tggttactca ccactgcgat ccccgggaaa acagcattcc aggtattaga
     4381 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     4441 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     4501 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     4561 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     4621 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     4681 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     4741 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     4801 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     4861 ctaagaatta attcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     4921 gggttccgcg cacatttccc cgaaaagtgc cacctgaaat tgtaaacgtt aatattttgt
     4981 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     5041 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     5101 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     5161 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     5221 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     5281 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     5341 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     5401 tacagggcgc gtcccattcg cca
    //

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