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pET-29b(+)

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  • Ybscience
  • 中国/美国
  • YB-0033
  • 2025年07月16日
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    上海钰博生物科技有限公司
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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-29b(+)

    • 库存：

      大量

    • 供应商：

      钰博生物

    • 规格：

      1ug/5ug

    pET-29b(+)

    pET29b载体基本信息

    出品公司: Ybscience
    别名: pET29b, pet 29b
    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: 5370 bp
    5' 测序引物: T7
    5' 测序引物序列:   5'-TAATACGACTCACTATAGGG-3'
    载体标签: N-S, N-Thrombin, C-His
    载体抗性: Kanamycin
    表达宿主菌: BL21(DE3) 、BLR(DE3)、BL21(DE3)pLysS
    备注: N term thrombin 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

    pET29b载体质粒图谱和多克隆位点信息

    pet29b载体图谱
    pet29b多克隆位点

    Feature Name Start End  
    T7_terminator 129 1
    T7_Terminal_primer 69 87
    lacI 217 1856
    T7_transl_en_RBS 320 304
    lacO 365 338
    T7_promoter 383 365
    tet (300 - 563) 419 682
    pBRrevBam_primer 490 471
    tet (576 - 851) 1915 2190
    ROP 2665 2856
    pGEX_3_primer 2872 2850
    pBR322_origin 3890 3271
    KanR2 3996 4811
    ORF Start End  
    ORF frame 2 296 1024
    ORF frame 3 897 1856
    ORF frame 3 3996 4811
    Enzyme Name Cut
    XhoI 158
    NotI 166
    EagI 166
    HindIII 173
    SalI 179
    SacI 190
    EcoRI 192
    BamHI 198
    EcoRV 206
    NcoI 211
    KpnI 237
    BglII 240
    BstBI 267
    NdeI 294
    XbaI 332
    BclI 1138
    ApaI 1335
    HpaI 1630
    FspI 2206
    NruI 4084
    SmaI 4301
    XmaI 4299
     

     

    pET29b载体简介

    The pET-29a-c(+) vectors carry an N-terminal S•Tag™/thrombin 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).

    pET29b载体序列

    LOCUS       pET-29 b (+)    5370 bp     DNA Circular    SYN
    DEFINITION  pET-29 b (+)
    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..5370
                         /organism="pET-29 b (+)"
                         /mol_type="other DNA"
         terminator      complement(1..129)
                         /label="T7_terminator"
         misc_feature    69..87
                         /label="T7_Terminal_primer"
         misc_feature    217..1856
                         /label="lacI"
         CDS             296..1024
                         /label="ORF frame 2"
                         /translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
                         IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
                         PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
                         ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
                         TVVADWRCHLQSGPARAVANCRGD*"
         misc_feature    complement(304..320)
                         /label="T7_transl_en_RBS"
                         /translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
                         IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
                         PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
                         ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
                         TVVADWRCHLQSGPARAVANCRGD*"
         misc_feature    complement(338..365)
                         /label="lacO"
                         /translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
                         IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
                         PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
                         ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
                         TVVADWRCHLQSGPARAVANCRGD*"
         promoter        complement(365..383)
                         /label="T7_promoter"
                         /translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
                         IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
                         PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
                         ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
                         TVVADWRCHLQSGPARAVANCRGD*"
         gene            419..682
                         /label="tet (300 - 563)"
                         /gene="tet (300 - 563)"
                         /translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
                         IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
                         PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
                         ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
                         TVVADWRCHLQSGPARAVANCRGD*"
         misc_feature    complement(471..490)
                         /label="pBRrevBam_primer"
                         /translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
                         IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
                         PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
                         ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
                         TVVADWRCHLQSGPARAVANCRGD*"
         CDS             897..1856
                         /label="ORF frame 3"
                         /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                         RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                         PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                         HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                         SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                         LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
         gene            1915..2190
                         /label="tet (576 - 851)"
                         /gene="tet (576 - 851)"
                         /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                         RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                         PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                         HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                         SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                         LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
         misc_feature    2665..2856
                         /label="ROP"
                         /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                         RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                         PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                         HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                         SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                         LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
         misc_feature    complement(2850..2872)
                         /label="pGEX_3_primer"
                         /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                         RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                         PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                         HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                         SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                         LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
         rep_origin      complement(3271..3890)
                         /label="pBR322_origin"
                         /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                         RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                         PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                         HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                         SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                         LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
         gene            3996..4811
                         /label="KanR2"
                         /gene="KanR2"
                         /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                         RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                         PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                         HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                         SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                         LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
         CDS             3996..4811
                         /label="ORF frame 3"
                         /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 tccgatatcg ccatggaacc gcgtggcacc agggtaccca
      241 gatctgggct gtccatgtgc tggcgttcga atttagcagc agcggtttct ttcatatgta
      301 tatctccttc ttaaagttaa acaaaattat ttctagaggg gaattgttat ccgctcacaa
      361 ttcccctata gtgagtcgta ttaatttcgc gggatcgaga tcgatctcga tcctctacgc
      421 cggacgcatc gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc
      481 cgacatcacc gatggggaag atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg
      541 cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg ggcgccatct ccttgcatgc
      601 accattcctt gcggcggcgg tgctcaacgg cctcaaccta ctactgggct gcttcctaat
      661 gcaggagtcg cataagggag agcgtcgaga tcccggacac catcgaatgg cgcaaaacct
      721 ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac
      781 cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg
      841 tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg
      901 cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc
      961 tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga
     1021 ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg
     1081 gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga
     1141 tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg
     1201 ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt attttctccc
     1261 atgaagacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg
     1321 cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata
     1381 aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca
     1441 tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc
     1501 tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc
     1561 gcgttggtgc ggacatctcg gtagtgggat acgacgatac cgaagacagc tcatgttata
     1621 tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc
     1681 gcttgctgca actctctcag ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac
     1741 tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg
     1801 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     1861 aacgcaatta atgtaagtta gctcactcat taggcaccgg gatctcgacc gatgcccttg
     1921 agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat cgtcgccgca
     1981 cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc gctctgggtc
     2041 attttcggcg aggaccgctt tcgctggagc gcgacgatga tcggcctgtc gcttgcggta
     2101 ttcggaatct tgcacgccct cgctcaagcc ttcgtcactg gtcccgccac caaacgtttc
     2161 ggcgagaagc aggccattat cgccggcatg gcggccccac gggtgcgcat gatcgtgctc
     2221 ctgtcgttga ggacccggct aggctggcgg ggttgcctta ctggttagca gaatgaatca
     2281 ccgatacgcg agcgaacgtg aagcgactgc tgctgcaaaa cgtctgcgac ctgagcaaca
     2341 acatgaatgg tcttcggttt ccgtgtttcg taaagtctgg aaacgcggaa gtcagcgccc
     2401 tgcaccatta tgttccggat ctgcatcgca ggatgctgct ggctaccctg tggaacacct
     2461 acatctgtat taacgaagcg ctggcattga ccctgagtga tttttctctg gtcccgccgc
     2521 atccataccg ccagttgttt accctcacaa cgttccagta accgggcatg ttcatcatca
     2581 gtaacccgta tcgtgagcat cctctctcgt ttcatcggta tcattacccc catgaacaga
     2641 aatccccctt acacggaggc atcagtgacc aaacaggaaa aaaccgccct taacatggcc
     2701 cgctttatca gaagccagac attaacgctt ctggagaaac tcaacgagct ggacgcggat
     2761 gaacaggcag acatctgtga atcgcttcac gaccacgctg atgagcttta ccgcagctgc
     2821 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     2881 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     2941 gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact
     3001 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatata tgcggtgtga
     3061 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg cttcctcgct
     3121 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     3181 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     3241 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     3301 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3361 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3421 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3481 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3541 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3601 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3661 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     3721 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     3781 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     3841 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     3901 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga acaataaaac
     3961 tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt
     4021 cttgctctag gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg
     4081 ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg attgtatggg aagcccgatg
     4141 cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga
     4201 tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc
     4261 gtactcctga tgatgcatgg ttactcacca ctgcgatccc cgggaaaaca gcattccagg
     4321 tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc
     4381 gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc
     4441 tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg
     4501 agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct
     4561 caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg
     4621 ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc
     4681 ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc
     4741 aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg
     4801 agtttttcta agaattaatt catgagcgga tacatatttg aatgtattta gaaaaataaa
     4861 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgaaattgt aaacgttaat
     4921 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     4981 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     5041 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     5101 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     5161 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     5221 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     5281 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     5341 gcgccgctac agggcgcgtc ccattcgcca
    //

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