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pET-30 Xa/LIC载体

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  • ZY69073-3
  • 2025年07月13日
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    • 详细信息
    • 文献和实验
    • 技术资料
    • 保存条件

      -20℃低温保存

    • 保质期

      三年

    • 英文名

      pET-30 Xa/LIC

    • 库存

      20

    • 供应商

      泽叶生物

    pET30 Xa/LIC载体基本信息

    出品公司: EMD Biosciences (Novagen)
    别名: pET30 Xa/LIC, pet 30a Xa/LIC
    质粒类型: 大肠杆菌蛋白表达
    表达水平:
    克隆方法: 多克隆位点,限制性内切酶
    载体大小: 5448 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-Xa Factor, C-His
    载体抗性: Kanamycin
    表达宿主菌: BL21(DE3) 、 Rosetta2(DE3)、C43(DE3)
    备注: For directional cloning of PCR-amplified DNA; ligation independent cloning; Factor Xa cleavage site
    产品目录号: ZY69073-3
    稳定性: 瞬时表达 Transient
    组成型: 组成型 Constitutive
    病毒/非病毒: 非病毒

    pET30 Xa/LIC载体质粒图谱和多克隆位点信息

    pet30 Xa/LIC载体图谱
    pet30 Xa/LIC多克隆位点
    pet30 Xa/LIC克隆方法

    Feature Name Start End
    T7_terminator 129 1
    T7_Terminal_primer 69 87
    lacI 328 1934
    T7_transl_en_RBS 398 382
    lacO 443 416
    T7_promoter 461 443
    tet (300 - 563) 497 760
    pBRrevBam_primer 568 549
    tet (576 - 851) 1993 2268
    ROP 2743 2934
    pGEX_3_primer 2950 2928
    pBR322_origin 3968 3349
    KanR2 4074 4889
    ORF Start End
    ORF frame 2 353 1102
    ORF frame 3 975 1934
    ORF frame 3 4074 4889
    Enzyme Name Cut
    XhoI 158
    NotI 166
    EagI 166
    HindIII 173
    SalI 179
    SacI 190
    EcoRI 192
    BamHI 198
    EcoRV 206
    NcoI 211
    MscI 216
    AgeI 257
    KpnI 264
    BglII 267
    BstBI 294
    NdeI 372
    XbaI 410
    BclI 1216
    ApaI 1413
    HpaI 1708
    FspI 2284
    NruI 4162
    SmaI 4379
    XmaI 4377

    pET30 Xa/LIC载体简介

    The pET-30 Xa/LIC vector is designed for rapid, directional cloning of PCR-amplified DNA for high-level expression of polypeptides containing N-terminal His•Tag® and S•Tag™ sequences for detection and purification. Using specifically designed primers for amplification and the pET-30Xa/LIC Vector Kit (Cat. No. 69073-3), inserts can be efficiently cloned without the need for restriction digestion or ligation, and the resulting fusion proteins can be cleaved with Factor Xa to precisely remove all vector-encoded amino acids. 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 circle map. The cloning/expression region of the coding strand transcribed by T7 RNA polymeraseis 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).

    pET30 Xa/LIC载体序列

    LOCUS pET-30 Xa/LIC 5448 bp DNA Circular SYN
    DEFINITION pET-30 Xa/LIC
    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..5448
    /organism="pET-30 Xa/LIC"
    /mol_type="other DNA"
    terminator complement(1..129)
    /label="T7_terminator"
    misc_feature 69..87
    /label="T7_Terminal_primer"
    misc_feature 328..1934
    /label="lacI"
    CDS 353..1102
    /label="ORF frame 2"
    /translation="MMMMMVHMYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSI
    SILYAGRIVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLL
    GAISLHAPFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRE
    SIQGGECETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPR
    GTTTGGQTVVADWRCHLQSGPARAVANCRGD*"
    misc_feature complement(382..398)
    /label="T7_transl_en_RBS"
    /translation="MMMMMVHMYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSI
    SILYAGRIVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLL
    GAISLHAPFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRE
    SIQGGECETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPR
    GTTTGGQTVVADWRCHLQSGPARAVANCRGD*"
    misc_feature complement(416..443)
    /label="lacO"
    /translation="MMMMMVHMYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSI
    SILYAGRIVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLL
    GAISLHAPFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRE
    SIQGGECETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPR
    GTTTGGQTVVADWRCHLQSGPARAVANCRGD*"
    promoter complement(443..461)
    /label="T7_promoter"
    /translation="MMMMMVHMYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSI
    SILYAGRIVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLL
    GAISLHAPFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRE
    SIQGGECETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPR
    GTTTGGQTVVADWRCHLQSGPARAVANCRGD*"
    gene 497..760
    /label="tet (300 - 563)"
    /gene="tet (300 - 563)"
    /translation="MMMMMVHMYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSI
    SILYAGRIVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLL
    GAISLHAPFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRE
    SIQGGECETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPR
    GTTTGGQTVVADWRCHLQSGPARAVANCRGD*"
    misc_feature complement(549..568)
    /label="pBRrevBam_primer"
    /translation="MMMMMVHMYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSI
    SILYAGRIVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLL
    GAISLHAPFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRE
    SIQGGECETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPR
    GTTTGGQTVVADWRCHLQSGPARAVANCRGD*"
    CDS 975..1934
    /label="ORF frame 3"
    /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
    RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
    PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
    HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
    SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
    LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
    gene 1993..2268
    /label="tet (576 - 851)"
    /gene="tet (576 - 851)"
    /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
    RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
    PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
    HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
    SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
    LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
    misc_feature 2743..2934
    /label="ROP"
    /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
    RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
    PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
    HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
    SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
    LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
    misc_feature complement(2928..2950)
    /label="pGEX_3_primer"
    /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
    RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
    PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
    HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
    SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
    LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
    rep_origin complement(3349..3968)
    /label="pBR322_origin"
    /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
    RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
    PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
    HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
    SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
    LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
    gene 4074..4889
    /label="KanR2"
    /gene="KanR2"
    /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
    RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
    PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
    HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
    SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
    LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
    CDS 4074..4889
    /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 ccatggccag aggagagtta gagcgaccct
    241 caataccgga gccaccaccg gtacccagat ctgggctgtc catgtgctgg cgttcgaatt
    301 tagcagcagc ggtttctttc ataccagaac cgcgtggcac cagaccagaa gaatgatgat
    361 gatgatggtg catatgtata tctccttctt aaagttaaac aaaattattt ctagagggga
    421 attgttatcc gctcacaatt cccctatagt gagtcgtatt aatttcgcgg gatcgagatc
    481 gatctcgatc ctctacgccg gacgcatcgt ggccggcatc accggcgcca caggtgcggt
    541 tgctggcgcc tatatcgccg acatcaccga tggggaagat cgggctcgcc acttcgggct
    601 catgagcgct tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg gactgttggg
    661 cgccatctcc ttgcatgcac cattccttgc ggcggcggtg ctcaacggcc tcaacctact
    721 actgggctgc ttcctaatgc aggagtcgca taagggagag cgtcgagatc ccggacacca
    781 tcgaatggcg caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca
    841 gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt
    901 atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa
    961 aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg
    1021 cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt
    1081 cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt
    1141 cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc
    1201 aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg
    1261 aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca
    1321 acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat ctggtcgcat
    1381 tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc
    1441 gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata gcggaacggg
    1501 aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg aatgagggca
    1561 tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca atgcgcgcca
    1621 ttaccgagtc cgggctgcgc gttggtgcgg acatctcggt agtgggatac gacgataccg
    1681 aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg
    1741 ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc
    1801 agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg
    1861 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
    1921 aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc tcactcatta ggcaccggga
    1981 tctcgaccga tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc
    2041 atgactatcg tcgccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg
    2101 ccggcagcgc tctgggtcat tttcggcgag gaccgctttc gctggagcgc gacgatgatc
    2161 ggcctgtcgc ttgcggtatt cggaatcttg cacgccctcg ctcaagcctt cgtcactggt
    2221 cccgccacca aacgtttcgg cgagaagcag gccattatcg ccggcatggc ggccccacgg
    2281 gtgcgcatga tcgtgctcct gtcgttgagg acccggctag gctggcgggg ttgccttact
    2341 ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg ctgcaaaacg
    2401 tctgcgacct gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta aagtctggaa
    2461 acgcggaagt cagcgccctg caccattatg ttccggatct gcatcgcagg atgctgctgg
    2521 ctaccctgtg gaacacctac atctgtatta acgaagcgct ggcattgacc ctgagtgatt
    2581 tttctctggt cccgccgcat ccataccgcc agttgtttac cctcacaacg ttccagtaac
    2641 cgggcatgtt catcatcagt aacccgtatc gtgagcatcc tctctcgttt catcggtatc
    2701 attaccccca tgaacagaaa tcccccttac acggaggcat cagtgaccaa acaggaaaaa
    2761 accgccctta acatggcccg ctttatcaga agccagacat taacgcttct ggagaaactc
    2821 aacgagctgg acgcggatga acaggcagac atctgtgaat cgcttcacga ccacgctgat
    2881 gagctttacc gcagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
    2941 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
    3001 cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc
    3061 gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
    3121 accatatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
    3181 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
    3241 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
    3301 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
    3361 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
    3421 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
    3481 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
    3541 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
    3601 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
    3661 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
    3721 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
    3781 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
    3841 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
    3901 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
    3961 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
    4021 ggtcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc
    4081 atattcaacg ggaaacgtct tgctctaggc cgcgattaaa ttccaacatg gatgctgatt
    4141 tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgat
    4201 tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca
    4261 atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga
    4321 ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg
    4381 ggaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg
    4441 cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca
    4501 gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg
    4561 cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc
    4621 ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata
    4681 accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg
    4741 cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat
    4801 tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt
    4861 ttcatttgat gctcgatgag tttttctaag aattaattca tgagcggata catatttgaa
    4921 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
    4981 gaaattgtaa acgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
    5041 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
    5101 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
    5161 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
    5221 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
    5281 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
    5341 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
    5401 acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgcca
    //

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