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pET-29a(+)

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  • Ybscience
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  • YB-00330
  • 2025年07月11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-29a(+)

    • 库存：

      大量

    • 供应商：

      钰博生物

    • 规格：

      1ug/5ug

    pET-29a(+)

    pET29a载体基本信息

    出品公司: Ybscience
    别名: pET29a, pet 29a
    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: 5371 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
     

    pET29a载体质粒图谱和多克隆位点信息

    pet29a载体图谱
    pet29a多克隆位点

    Feature Name Start End  
    T7_terminator 129 1
    T7_Terminal_primer 69 87
    lacI 218 1857
    T7_transl_en_RBS 321 305
    lacO 366 339
    T7_promoter 384 366
    tet (300 - 563) 420 683
    pBRrevBam_primer 491 472
    tet (576 - 851) 1916 2191
    ROP 2666 2857
    pGEX_3_primer 2873 2851
    pBR322_origin 3891 3272
    KanR2 3997 4812
    ORF Start End  
    ORF frame 3 297 1025
    ORF frame 1 898 1857
    ORF frame 1 3997 4812
    Enzyme Name Cut
    XhoI 158
    NotI 166
    EagI 166
    HindIII 173
    SalI 179
    SacI 190
    EcoRI 192
    BamHI 198
    EcoRV 206
    NcoI 212
    KpnI 238
    BglII 241
    BstBI 268
    NdeI 295
    XbaI 333
    BclI 1139
    ApaI 1336
    HpaI 1631
    FspI 2207
    NruI 4085
    SmaI 4302
    XmaI 4300
     

     

    pET29a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).

    pET29a载体序列

    LOCUS       pET-29 a (+)    5371 bp     DNA  Circular   SYN
    DEFINITION  pET-29 a (+)
    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..5371
                         /organism="pET-29 a (+)"
                         /mol_type="other DNA"
         terminator      complement(1..129)
                         /label="T7_terminator"
         misc_feature    69..87
                         /label="T7_Terminal_primer"
         misc_feature    218..1857
                         /label="lacI"
         CDS             297..1025
                         /label="ORF frame 3"
                         /translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
                         IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
                         PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
                         ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
                         TVVADWRCHLQSGPARAVANCRGD*"
         misc_feature    complement(305..321)
                         /label="T7_transl_en_RBS"
                         /translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
                         IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
                         PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
                         ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
                         TVVADWRCHLQSGPARAVANCRGD*"
         misc_feature    complement(339..366)
                         /label="lacO"
                         /translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
                         IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
                         PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
                         ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
                         TVVADWRCHLQSGPARAVANCRGD*"
         promoter        complement(366..384)
                         /label="T7_promoter"
                         /translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
                         IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
                         PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
                         ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
                         TVVADWRCHLQSGPARAVANCRGD*"
         gene            420..683
                         /label="tet (300 - 563)"
                         /gene="tet (300 - 563)"
                         /translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
                         IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
                         PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
                         ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
                         TVVADWRCHLQSGPARAVANCRGD*"
         misc_feature    complement(472..491)
                         /label="pBRrevBam_primer"
                         /translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
                         IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
                         PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
                         ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
                         TVVADWRCHLQSGPARAVANCRGD*"
         CDS             898..1857
                         /label="ORF frame 1"
                         /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                         RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                         PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                         HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                         SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                         LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
         gene            1916..2191
                         /label="tet (576 - 851)"
                         /gene="tet (576 - 851)"
                         /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                         RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                         PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                         HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                         SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                         LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
         misc_feature    2666..2857
                         /label="ROP"
                         /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                         RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                         PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                         HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                         SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                         LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
         misc_feature    complement(2851..2873)
                         /label="pGEX_3_primer"
                         /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                         RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                         PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                         HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                         SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                         LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
         rep_origin      complement(3272..3891)
                         /label="pBR322_origin"
                         /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                         RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                         PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                         HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                         SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                         LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
         gene            3997..4812
                         /label="KanR2"
                         /gene="KanR2"
                         /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                         RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                         PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                         HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                         SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                         LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
         CDS             3997..4812
                         /label="ORF frame 1"
                         /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 tccgatatca gccatggaac cgcgtggcac cagggtaccc
      241 agatctgggc tgtccatgtg ctggcgttcg aatttagcag cagcggtttc tttcatatgt
      301 atatctcctt cttaaagtta aacaaaatta tttctagagg ggaattgtta tccgctcaca
      361 attcccctat agtgagtcgt attaatttcg cgggatcgag atcgatctcg atcctctacg
      421 ccggacgcat cgtggccggc atcaccggcg ccacaggtgc ggttgctggc gcctatatcg
      481 ccgacatcac cgatggggaa gatcgggctc gccacttcgg gctcatgagc gcttgtttcg
      541 gcgtgggtat ggtggcaggc cccgtggccg ggggactgtt gggcgccatc tccttgcatg
      601 caccattcct tgcggcggcg gtgctcaacg gcctcaacct actactgggc tgcttcctaa
      661 tgcaggagtc gcataaggga gagcgtcgag atcccggaca ccatcgaatg gcgcaaaacc
      721 tttcgcggta tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa
      781 ccagtaacgt tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc
      841 gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg
      901 gcggagctga attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg
      961 ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg
     1021 attaaatctc gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc
     1081 ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg
     1141 atcattaact atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat
     1201 gttccggcgt tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc
     1261 catgaagacg gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc
     1321 gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat
     1381 aaatatctca ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc
     1441 atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg
     1501 ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg
     1561 cgcgttggtg cggacatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat
     1621 atcccgccgt taaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac
     1681 cgcttgctgc aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca
     1741 ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     1801 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     1861 caacgcaatt aatgtaagtt agctcactca ttaggcaccg ggatctcgac cgatgccctt
     1921 gagagccttc aacccagtca gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc
     1981 acttatgact gtcttcttta tcatgcaact cgtaggacag gtgccggcag cgctctgggt
     2041 cattttcggc gaggaccgct ttcgctggag cgcgacgatg atcggcctgt cgcttgcggt
     2101 attcggaatc ttgcacgccc tcgctcaagc cttcgtcact ggtcccgcca ccaaacgttt
     2161 cggcgagaag caggccatta tcgccggcat ggcggcccca cgggtgcgca tgatcgtgct
     2221 cctgtcgttg aggacccggc taggctggcg gggttgcctt actggttagc agaatgaatc
     2281 accgatacgc gagcgaacgt gaagcgactg ctgctgcaaa acgtctgcga cctgagcaac
     2341 aacatgaatg gtcttcggtt tccgtgtttc gtaaagtctg gaaacgcgga agtcagcgcc
     2401 ctgcaccatt atgttccgga tctgcatcgc aggatgctgc tggctaccct gtggaacacc
     2461 tacatctgta ttaacgaagc gctggcattg accctgagtg atttttctct ggtcccgccg
     2521 catccatacc gccagttgtt taccctcaca acgttccagt aaccgggcat gttcatcatc
     2581 agtaacccgt atcgtgagca tcctctctcg tttcatcggt atcattaccc ccatgaacag
     2641 aaatccccct tacacggagg catcagtgac caaacaggaa aaaaccgccc ttaacatggc
     2701 ccgctttatc agaagccaga cattaacgct tctggagaaa ctcaacgagc tggacgcgga
     2761 tgaacaggca gacatctgtg aatcgcttca cgaccacgct gatgagcttt accgcagctg
     2821 cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     2881 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     2941 tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtatac
     3001 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat atgcggtgtg
     3061 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc
     3121 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     3181 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     3241 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     3301 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     3361 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     3421 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     3481 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     3541 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3601 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     3661 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     3721 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     3781 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     3841 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     3901 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg aacaataaaa
     3961 ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg
     4021 tcttgctcta ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg
     4081 gctcgcgata atgtcgggca atcaggtgcg acaatctatc gattgtatgg gaagcccgat
     4141 gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag
     4201 atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc
     4261 cgtactcctg atgatgcatg gttactcacc actgcgatcc ccgggaaaac agcattccag
     4321 gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg
     4381 cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt
     4441 ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac
     4501 gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc
     4561 tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag
     4621 gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat
     4681 cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt
     4741 caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat
     4801 gagtttttct aagaattaat tcatgagcgg atacatattt gaatgtattt agaaaaataa
     4861 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgaaattg taaacgttaa
     4921 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     4981 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     5041 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     5101 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     5161 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     5221 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     5281 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     5341 tgcgccgcta cagggcgcgt cccattcgcc a
    //

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