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pTYB1

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  • Ybscience
  • 中国/美国
  • YB-0147
  • 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

      pTYB1

    • 库存

      大量

    • 供应商

      钰博生物

    • 规格

      1ug/5ug

    pTYB1

    载体基本信息

    出品公司: Ybscience
      pTYB1
    质粒类型: 大肠杆菌表达载体;蛋白纯化
    高拷贝/低拷贝: --
    启动子: T7
    克隆方法: 多克隆位点,限制性内切酶
    载体大小: 7477 bp
    5' 测序引物及序列: T7: TAATACGACTCACTATAGGG
    3' 测序引物及序列: --
    载体标签: Intein
    载体抗性: Ampicillin
    筛选标记: --
    备注: --
    稳定性: --
    组成型: --
    病毒/非病毒: 非病毒
     

    载体质粒图谱和多克隆位点信息

    pTYB1载体图谱

    pTYB1多克隆位点

    pTYB1载体特征

    载体简介

     

    pTYB1 is an E. coli plasmid cloning vector designed for recombinant protein expression and purification using the 
    IMPACT? Kit (NEB #E6901) (1,2). It contains the pMB1 origin of replication from pBR322 and is maintained at a similar 
    copy number to pBR322; in addition, pTYB1 also contains an M13 origin of replication.
    The multiple cloning site (MCS) is positioned to allow translational fusion of the Sce VMA intein tag to the C-terminus 
    of the cloned target protein (1). The chitin binding domain (CBD) from B. circulans, fused to the C-terminus of the intein, 
    facilitates purification of the intein-target protein precursor.
    Transcription of the gene fusion is controlled by the inducible T7 promoter, requiring E. coli strains containing integrated 
    copies of the T7 RNA polymerase gene [e.g., C2566, C2833 or BL21(DE3)] for expression. Basal expression from the T7 promoter 
    is minimized by the binding of the Lac repressor, encoded by the lacI gene, to the lac operator immediately downstream of the T7 promoter (3). Translation of the fusion utilizes the translation initiation signal (Shine Dalgarno sequence) from the 
    strongly expressed T7 gene 10 protein (φ10).
    pTYB1, pTYB2, pTYB3, and pTYB4 are identical except for the MCS regions (opposite page). All four vectors contain either an 
    NdeI (pTYB1 and pTYB2) or NcoI (pTYB3 and pTYB4) siteoverlapping the initiating methionine codon of the intein fusion gene. 
    The N-terminal cysteine residue (“Cys1”) of the intein is shaded.
    Enzymes with unique restriction sites are shown in bold type. Location of sites of all NEB restriction enzymes can be found 
    on the NEB web site (choose Technical Reference > DNA Sequences and Maps). Restriction site coordinates refer to the position of the 5′-most base on the top strand in each recognition sequence.
    Open reading frame (ORF) coordinates are in the form "translational start – translational stop”; numbers refer to positions on the top (clockwise) strand, regardless of the direction of transcription and include the start and stop codons. Component 
    genes or regions of fusion ORFs are indented below the ORF itself.
    pMB1 origin of replication coordinates include the region from the -35 promoter sequence of the RNAII transcript to the 
    RNA/DNA switch point. For the M13 origin, the arrow shows the direction of synthesis of the (+) strand, which gets packaged 
    into phage particles. bla (ApR) gene coordinates include the signal sequence.
    
    

     

    载体序列

    LOCUS       pTYB1	7477 bp 	DNA	SYN
    DEFINITION  pTYB1
    ACCESSION   
    KEYWORDS    
    SOURCE      
      ORGANISM  other sequences; artificial sequences; vectors.
    FEATURES             Location/Qualifiers
         source          1..7477
                         /organism="pTYB1"
                         /mol_type="other DNA"
         promoter        70..98
                         /label="AmpR_promoter"
         gene            140..1000
                         /label="Ampicillin"
                         /gene="Ampicillin"
         CDS             140..1000
                         /label="ORF frame 2"
                         /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                         IELDLNSGKILESFRPEERSPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
                         YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                         DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                         LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                         EIGASLIKHW*"
         rep_origin      complement(1232..1538)
                         /label="f1_origin"
                         /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                         IELDLNSGKILESFRPEERSPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
                         YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                         DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                         LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                         EIGASLIKHW*"
         rep_origin      1650..2269
                         /label="pBR322_origin"
                         /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                         IELDLNSGKILESFRPEERSPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
                         YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                         DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                         LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                         EIGASLIKHW*"
         misc_feature    2607..2629
                         /label="pGEX_3_primer"
                         /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                         IELDLNSGKILESFRPEERSPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
                         YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                         DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                         LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                         EIGASLIKHW*"
         misc_feature    complement(2623..2814)
                         /label="ROP"
                         /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                         IELDLNSGKILESFRPEERSPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
                         YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                         DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                         LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                         EIGASLIKHW*"
         CDS             2953..3588
                         /label="ORF frame 1"
                         /translation="MDAAGPEKNHSGSMPAERQQDVAQRVGRHAGDNGLLLAETFGGG
                         TSDEGLSEGVQDSEYRKRQADHRRAPAKAVLAENDPERCRHLSYELHDKEDSHKCGDD
                         SHAPRPPEGADWVEGSQGHRSRSRCLMSELTYINCVALTARFPVGKPVVPAALMNRPT
                         RGERRFAYWAPGWFFFSPVRRATADCPSPPGPERVAASGPRWFAPAGENPV*"
         gene            complement(3010..3309)
                         /label="tet (611 - 312)"
                         /gene="tet (611 - 312)"
                         /translation="MDAAGPEKNHSGSMPAERQQDVAQRVGRHAGDNGLLLAETFGGG
                         TSDEGLSEGVQDSEYRKRQADHRRAPAKAVLAENDPERCRHLSYELHDKEDSHKCGDD
                         SHAPRPPEGADWVEGSQGHRSRSRCLMSELTYINCVALTARFPVGKPVVPAALMNRPT
                         RGERRFAYWAPGWFFFSPVRRATADCPSPPGPERVAASGPRWFAPAGENPV*"
         misc_feature    complement(3371..4462)
                         /label="lacI"
                         /translation="MDAAGPEKNHSGSMPAERQQDVAQRVGRHAGDNGLLLAETFGGG
                         TSDEGLSEGVQDSEYRKRQADHRRAPAKAVLAENDPERCRHLSYELHDKEDSHKCGDD
                         SHAPRPPEGADWVEGSQGHRSRSRCLMSELTYINCVALTARFPVGKPVVPAALMNRPT
                         RGERRFAYWAPGWFFFSPVRRATADCPSPPGPERVAASGPRWFAPAGENPV*"
         CDS             complement(3371..4330)
                         /label="ORF frame 1"
                         /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                         RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                         PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                         HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                         SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                         LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
         CDS             complement(4654..5415)
                         /label="ORF frame 2"
                         /translation="MPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVHRQT
                         TDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVH
                         RQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQE
                         SVHRQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICP
                         TQESVHRQTTDKTKGPVFRLSLSFYLMPGELGINS*"
         terminator      4687..4730
                         /label="rrnB_T1_terminator"
                         /translation="MPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVHRQT
                         TDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVH
                         RQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQE
                         SVHRQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICP
                         TQESVHRQTTDKTKGPVFRLSLSFYLMPGELGINS*"
         terminator      4870..4913
                         /label="rrnB_T1_terminator"
                         /translation="MPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVHRQT
                         TDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVH
                         RQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQE
                         SVHRQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICP
                         TQESVHRQTTDKTKGPVFRLSLSFYLMPGELGINS*"
         terminator      5053..5096
                         /label="rrnB_T1_terminator"
                         /translation="MPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVHRQT
                         TDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVH
                         RQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQE
                         SVHRQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICP
                         TQESVHRQTTDKTKGPVFRLSLSFYLMPGELGINS*"
         terminator      5236..5279
                         /label="rrnB_T1_terminator"
                         /translation="MPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVHRQT
                         TDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVH
                         RQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQE
                         SVHRQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICP
                         TQESVHRQTTDKTKGPVFRLSLSFYLMPGELGINS*"
         terminator      5419..5462
                         /label="rrnB_T1_terminator"
                         /translation="MPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVHRQT
                         TDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVH
                         RQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQE
                         SVHRQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICP
                         TQESVHRQTTDKTKGPVFRLSLSFYLMPGELGINS*"
         promoter        5637..5655
                         /label="T7_promoter"
                         /translation="MPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVHRQT
                         TDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVH
                         RQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQE
                         SVHRQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICP
                         TQESVHRQTTDKTKGPVFRLSLSFYLMPGELGINS*"
         misc_feature    5655..5682
                         /label="lacO"
                         /translation="MPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVHRQT
                         TDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVH
                         RQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQE
                         SVHRQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICP
                         TQESVHRQTTDKTKGPVFRLSLSFYLMPGELGINS*"
         misc_feature    5700..5716
                         /label="T7_transl_en_RBS"
                         /translation="MPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVHRQT
                         TDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVH
                         RQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQE
                         SVHRQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICP
                         TQESVHRQTTDKTKGPVFRLSLSFYLMPGELGINS*"
         CDS             5725..7329
                         /label="ORF frame 1"
                         /translation="MASSRVDGGREFLEGSSCFAKGTNVLMADGSIECIENIEVGNKV
                         MGKDGRPREVIKLPRGRETMYSVVQKSQHRAHKSDSSREVPELLKFTCNATHELVVRT
                         PRSVRRLSRTIKGVEYFEVITFEMGQKKAPDGRIVELVKEVSKSYPISEGPERANELV
                         ESYRKASNKAYFEWTIEARDLSLLGSHVRKATYQTYAPILYENDHFFDYMQKSKFHLT
                         IEGPKVLAYLLGLWIGDGLSDRATFSVDSRDTSLMERVTEYAEKLNLCAEYKDRKEPQ
                         VAKTVNLYSKVVRGNGIRNNLNTENPLWDAIVGLGFLKDGVKNIPSFLSTDNIGTRET
                         FLAGLIDSDGYVTDEHGIKATIKTIHTSVRDGLVSLARSLGLVVSVNAEPAKVDMNVT
                         KHKISYAIYMSGGDVLLNVLSKCAGSKKFRPAPAAAFARECRGFYFELQELKEDDYYG
                         ITLSDDSDHQFLLGSQVVVHACGGLTGLNSGLTTNPGVSAWQVNTAYTAGQLVTYNGK
                         TYKCLQPHTSLAGWEPSNVPALWQLQ*"
         terminator      7349..7477
                         /label="T7_terminator"
                         /translation="MASSRVDGGREFLEGSSCFAKGTNVLMADGSIECIENIEVGNKV
                         MGKDGRPREVIKLPRGRETMYSVVQKSQHRAHKSDSSREVPELLKFTCNATHELVVRT
                         PRSVRRLSRTIKGVEYFEVITFEMGQKKAPDGRIVELVKEVSKSYPISEGPERANELV
                         ESYRKASNKAYFEWTIEARDLSLLGSHVRKATYQTYAPILYENDHFFDYMQKSKFHLT
                         IEGPKVLAYLLGLWIGDGLSDRATFSVDSRDTSLMERVTEYAEKLNLCAEYKDRKEPQ
                         VAKTVNLYSKVVRGNGIRNNLNTENPLWDAIVGLGFLKDGVKNIPSFLSTDNIGTRET
                         FLAGLIDSDGYVTDEHGIKATIKTIHTSVRDGLVSLARSLGLVVSVNAEPAKVDMNVT
                         KHKISYAIYMSGGDVLLNVLSKCAGSKKFRPAPAAAFARECRGFYFELQELKEDDYYG
                         ITLSDDSDHQFLLGSQVVVHACGGLTGLNSGLTTNPGVSAWQVNTAYTAGQLVTYNGK
                         TYKCLQPHTSLAGWEPSNVPALWQLQ*"
         misc_feature    complement(7391..7409)
                         /label="T7_Terminal_primer"
                         /translation="MASSRVDGGREFLEGSSCFAKGTNVLMADGSIECIENIEVGNKV
                         MGKDGRPREVIKLPRGRETMYSVVQKSQHRAHKSDSSREVPELLKFTCNATHELVVRT
                         PRSVRRLSRTIKGVEYFEVITFEMGQKKAPDGRIVELVKEVSKSYPISEGPERANELV
                         ESYRKASNKAYFEWTIEARDLSLLGSHVRKATYQTYAPILYENDHFFDYMQKSKFHLT
                         IEGPKVLAYLLGLWIGDGLSDRATFSVDSRDTSLMERVTEYAEKLNLCAEYKDRKEPQ
                         VAKTVNLYSKVVRGNGIRNNLNTENPLWDAIVGLGFLKDGVKNIPSFLSTDNIGTRET
                         FLAGLIDSDGYVTDEHGIKATIKTIHTSVRDGLVSLARSLGLVVSVNAEPAKVDMNVT
                         KHKISYAIYMSGGDVLLNVLSKCAGSKKFRPAPAAAFARECRGFYFELQELKEDDYYG
                         ITLSDDSDHQFLLGSQVVVHACGGLTGLNSGLTTNPGVSAWQVNTAYTAGQLVTYNGK
                         TYKCLQPHTSLAGWEPSNVPALWQLQ*"
    ORIGIN
        1 AACTACGTCA GGTGGCACTT TTCGGGGAAA TGTGCGCGGA ACCCCTATTT GTTTATTTTT
       61 CTAAATACAT TCAAATATGT ATCCGCTCAT GAGACAATAA CCCTGATAAA TGCTTCAATA
      121 ATATTGAAAA AGGAAGAGTA TGAGTATTCA ACATTTCCGT GTCGCCCTTA TTCCCTTTTT
      181 TGCGGCATTT TGCCTTCCTG TTTTTGCTCA CCCAGAAACG CTGGTGAAAG TAAAAGATGC
      241 TGAAGATCAG TTGGGTGCAC GAGTGGGTTA CATCGAACTG GATCTCAACA GCGGTAAGAT
      301 CCTTGAGAGT TTTCGCCCCG AAGAACGTTC TCCAATGATG AGCACTTTTA AAGTTCTGCT
      361 ATGTGGCGCG GTATTATCCC GTGTTGACGC CGGGCAAGAG CAACTCGGTC GCCGCATACA
      421 CTATTCTCAG AATGACTTGG TTGAGTACTC ACCAGTCACA GAAAAGCATC TTACGGATGG
      481 CATGACAGTA AGAGAATTAT GCAGTGCTGC CATAACCATG AGTGATAACA CTGCGGCCAA
      541 CTTACTTCTG ACAACGATCG GAGGACCGAA GGAGCTAACC GCTTTTTTGC ACAACATGGG
      601 GGATCATGTA ACTCGCCTTG ATCGTTGGGA ACCGGAGCTG AATGAAGCCA TACCAAACGA
      661 CGAGCGTGAC ACCACGATGC CTGTAGCAAT GGCAACAACG TTGCGCAAAC TATTAACTGG
      721 CGAACTACTT ACTCTAGCTT CCCGGCAACA ATTAATAGAC TGGATGGAGG CGGATAAAGT
      781 TGCAGGACCA CTTCTGCGCT CGGCCCTTCC GGCTGGCTGG TTTATTGCTG ATAAATCTGG
      841 AGCCGGTGAG CGTGGGTCTC GCGGTATCAT TGCAGCACTG GGGCCAGATG GTAAGCCCTC
      901 CCGTATCGTA GTTATCTACA CGACGGGGAG TCAGGCAACT ATGGATGAAC GAAATAGACA
      961 GATCGCTGAG ATAGGTGCCT CACTGATTAA GCATTGGTAA CTGTCAGACC AAGTTTACTC
     1021 ATATATACTT TAGATTGATT TACCCCGGTT GATAATCAGA AAAGCCCCAA AAACAGGAAG
     1081 ATTGTATAAG CAAATATTTA AATTGTAAAC GTTAATATTT TGTTAAAATT CGCGTTAAAT
     1141 TTTTGTTAAA TCAGCTCATT TTTTAACCAA TAGGCCGAAA TCGGCAAAAT CCCTTATAAA
     1201 TCAAAAGAAT AGCCCGAGAT AGGGTTGAGT GTTGTTCCAG TTTGGAACAA GAGTCCACTA
     1261 TTAAAGAACG TGGACTCCAA CGTCAAAGGG CGAAAAACCG TCTATCAGGG CGATGGCCCA
     1321 CTACGTGAAC CATCACCCAA ATCAAGTTTT TTGGGGTCGA GGTGCCGTAA AGCACTAAAT
     1381 CGGAACCCTA AAGGGAGCCC CCGATTTAGA GCTTGACGGG GAAAGCCGGC GAACGTGGCG
     1441 AGAAAGGAAG GGAAGAAAGC GAAAGGAGCG GGCGCTAGGG CGCTGGCAAG TGTAGCGGTC
     1501 ACGCTGCGCG TAACCACCAC ACCCGCCGCG CTTAATGCGC CGCTACAGGG CGCGTAAAAG
     1561 GATCTAGGTG AAGATCCTTT TTGATAATCT CATGACCAAA ATCCCTTAAC GTGAGTTTTC
     1621 GTTCCACTGA GCGTCAGACC CCGTAGAAAA GATCAAAGGA TCTTCTTGAG ATCCTTTTTT
     1681 TCTGCGCGTA ATCTGCTGCT TGCAAACAAA AAAACCACCG CTACCAGCGG TGGTTTGTTT
     1741 GCCGGATCAA GAGCTACCAA CTCTTTTTCC GAAGGTAACT GGCTTCAGCA GAGCGCAGAT
     1801 ACCAAATACT GTCCTTCTAG TGTAGCCGTA GTTAGGCCAC CACTTCAAGA ACTCTGTAGC
     1861 ACCGCCTACA TACCTCGCTC TGCTAATCCT GTTACCAGTG GCTGCTGCCA GTGGCGATAA
     1921 GTCGTGTCTT ACCGGGTTGG ACTCAAGACG ATAGTTACCG GATAAGGCGC AGCGGTCGGG
     1981 CTGAACGGGG GGTTCGTGCA CACAGCCCAG CTTGGAGCGA ACGACCTACA CCGAACTGAG
     2041 ATACCTACAG CGTGAGCTAT GAGAAAGCGC CACGCTTCCC GAAGGGAGAA AGGCGGACAG
     2101 GTATCCGGTA AGCGGCAGGG TCGGAACAGG AGAGCGCACG AGGGAGCTTC CAGGGGGAAA
     2161 CGCCTGGTAT CTTTATAGTC CTGTCGGGTT TCGCCACCTC TGACTTGAGC GTCGATTTTT
     2221 GTGATGCTCG TCAGGGGGGC GGAGCCTATG GAAAAACGCC AGCAACGCGG CCTTTTTACG
     2281 GTTCCTGGCC TTTTGCTGGC CTTTTGCTCA CATGTTCTTT CCTGCGTTAT CCCCTGATTC
     2341 TGTGGATAAC CGTATTACCG CCTTTGAGTG AGCTGATACC GCTCGCCGCA GCCGAACGAC
     2401 CGAGCGCAGC GAGTCAGTGA GCGAGGAAGC TATGGTGCAC TCTCAGTACA ATCTGCTCTG
     2461 ATGCCGCATA GTTAAGCCAG TATACACTCC GCTATCGCTA CGTGACTGGG TCATGGCTGC
     2521 GCCCCGACAC CCGCCAACAC CCGCTGACGC GCCCTGACGG GCTTGTCTGC TCCCGGCATC
     2581 CGCTTACAGA CAAGCTGTGA CCGTCTCCGG GAGCTGCATG TGTCAGAGGT TTTCACCGTC
     2641 ATCACCGAAA CGCGCGAGGC AGCTGCGGTA AAGCTCATCA GCGTGGTCGT GCAGCGATTC
     2701 ACAGATGTCT GCCTGTTCAT CCGCGTCCAG CTCGTTGAGT TTCTCCAGAA GCGTTAATGT
     2761 CTGGCTTCTG ATAAAGCGGG CCATGTTAAG GGCGGTTTTT TCCTGTTTGG TCACTGATGC
     2821 CTCCGTGTAA GGGGGATTTC TGTTCATGGG GGTAATGATA CCGATGAAAC GAGAGAGGAT
     2881 GCTCACGATA CGGGTTACTG ATGATGAACA TGCCCGGTTA CTGGAACGTT GTGAGGGTAA
     2941 ACAACTGGCG GTATGGATGC GGCGGGACCA GAGAAAAATC ACTCAGGGTC AATGCCAGCC
     3001 GAACGCCAGC AAGACGTAGC CCAGCGCGTC GGCCGCCATG CCGGCGATAA TGGCCTGCTT
     3061 CTCGCCGAAA CGTTTGGTGG CGGGACCAGT GACGAAGGCT TGAGCGAGGG CGTGCAAGAT
     3121 TCCGAATACC GCAAGCGACA GGCCGATCAT CGTCGCGCTC CAGCGAAAGC GGTCCTCGCC
     3181 GAAAATGACC CAGAGCGCTG CCGGCACCTG TCCTACGAGT TGCATGATAA AGAAGACAGT
     3241 CATAAGTGCG GCGACGATAG TCATGCCCCG CGCCCACCGG AAGGAGCTGA CTGGGTTGAA
     3301 GGCTCTCAAG GGCATCGGTC GAGATCCCGG TGCCTAATGA GTGAGCTAAC TTACATTAAT
     3361 TGCGTTGCGC TCACTGCCCG CTTTCCAGTC GGGAAACCTG TCGTGCCAGC TGCATTAATG
     3421 AATCGGCCAA CGCGCGGGGA GAGGCGGTTT GCGTATTGGG CGCCAGGGTG GTTTTTCTTT
     3481 TCACCAGTGA GACGGGCAAC AGCTGATTGC CCTTCACCGC CTGGCCCTGA GAGAGTTGCA
     3541 GCAAGCGGTC CACGCTGGTT TGCCCCAGCA GGCGAAAATC CTGTTTGATG GTGGTTAACG
     3601 GCGGGATATA ACATGAGCTG TCTTCGGTAT CGTCGTATCC CACTACCGAG ATATCCGCAC
     3661 CAACGCGCAG CCCGGACTCG GTAATGGCGC GCATTGCGCC CAGCGCCATC TGATCGTTGG
     3721 CAACCAGCAT CGCAGTGGGA ACGATGCCCT CATTCAGCAT TTGCATGGTT TGTTGAAAAC
     3781 CGGACATGGC ACTCCAGTCG CCTTCCCGTT CCGCTATCGG CTGAATTTGA TTGCGAGTGA
     3841 GATATTTATG CCAGCCAGCC AGACGCAGAC GCGCCGAGAC AGAACTTAAT GGGCCCGCTA
     3901 ACAGCGCGAT TTGCTGGTGA CCCAATGCGA CCAGATGCTC CACGCCCAGT CGCGTACCGT
     3961 CTTCATGGGA GAAAATAATA CTGTTGATGG GTGTCTGGTC AGAGACATCA AGAAATAACG
     4021 CCGGAACATT AGTGCAGGCA GCTTCCACAG CAATGGCATC CTGGTCATCC AGCGGATAGT
     4081 TAATGATCAG CCCACTGACG CGTTGCGCGA GAAGATTGTG CACCGCCGCT TTACAGGCTT
     4141 CGACGCCGCT TCGTTCTACC ATCGACACCA CCACGCTGGC ACCCAGTTGA TCGGCGCGAG
     4201 ATTTAATCGC CGCGACAATT TGCGACGGCG CGTGCAGGGC CAGACTGGAG GTGGCAACGC
     4261 CAATCAGCAA CGACTGTTTG CCCGCCAGTT GTTGTGCCAC GCGGTTGGGA ATGTAATTCA
     4321 GCTCCGCCAT CGCCGCTTCC ACTTTTTCCC GCGTTTTCGC AGAAACGTGG CTGGCCTGGT
     4381 TCACCACGCG GGAAACGGTC TGATAAGAGA CACCGGCATA CTCTGCGACA TCGTATAACG
     4441 TTACTGGTTT CACATTCACC ACCCTGAATT GACTCTCTTC CGGGCGCTAT CATGCCATAC
     4501 CGCGAAAGGT TTTGCGCCAT TCGATGGTGT CCGGGATCTC GACGCTCTCC CTTATGCGAC
     4561 TCCTGCATTA GGAAGCAGCC CAGTAGTAGG TTGAGGCCGT TGAGCACCGC CGCCGCAAGG
     4621 AATGGTGCAT GCCGGCATGC CGCCCTTTCG TCTTCAAGAA TTAATTCCCA ATTCCCCAGG
     4681 CATCAAATAA AACGAAAGGC TCAGTCGAAA GACTGGGCCT TTCGTTTTAT CTGTTGTTTG
     4741 TCGGTGAACG CTCTCCTGAG TAGGACAAAT CCGCCGGGAG CGGATTTGAA CGTTGCGAAG
     4801 CAACGGCCCG GAGGGTGGCG GGCAGGACGC CCGCCATAAA CTGCCAGGAA TTAATTCCCC
     4861 AGGCATCAAA TAAAACGAAA GGCTCAGTCG AAAGACTGGG CCTTTCGTTT TATCTGTTGT
     4921 TTGTCGGTGA ACGCTCTCCT GAGTAGGACA AATCCGCCGG GAGCGGATTT GAACGTTGCG
     4981 AAGCAACGGC CCGGAGGGTG GCGGGCAGGA CGCCCGCCAT AAACTGCCAG GAATTAATTC
     5041 CCCAGGCATC AAATAAAACG AAAGGCTCAG TCGAAAGACT GGGCCTTTCG TTTTATCTGT
     5101 TGTTTGTCGG TGAACGCTCT CCTGAGTAGG ACAAATCCGC CGGGAGCGGA TTTGAACGTT
     5161 GCGAAGCAAC GGCCCGGAGG GTGGCGGGCA GGACGCCCGC CATAAACTGC CAGGAATTAA
     5221 TTCCCCAGGC ATCAAATAAA ACGAAAGGCT CAGTCGAAAG ACTGGGCCTT TCGTTTTATC
     5281 TGTTGTTTGT CGGTGAACGC TCTCCTGAGT AGGACAAATC CGCCGGGAGC GGATTTGAAC
     5341 GTTGCGAAGC AACGGCCCGG AGGGTGGCGG GCAGGACGCC CGCCATAAAC TGCCAGGAAT
     5401 TAATTCCCCA GGCATCAAAT AAAACGAAAG GCTCAGTCGA AAGACTGGGC CTTTCGTTTT
     5461 ATCTGTTGTT TGTCGGTGAA CGCTCTCCTG AGTAGGACAA ATCCGCCGGG AGCGGATTTG
     5521 AACGTTGCGA AGCAACGGCC CGGAGGGTGG CGGGCAGGAC GCCCGCCATA AACTGCCAGG
     5581 AATTGGGGAT CGGAATTAAT TCCCGGTTTA AACCGGGGAT CTCGATCCCG CGAAATTAAT
     5641 ACGACTCACT ATAGGGGAAT TGTGAGCGGA TAACAATTCC CCTCTAGAAA TAATTTTGTT
     5701 TAACTTTAAG AAGGAGATAT ACATATGGCT AGCTCGCGAG TCGACGGCGG CCGCGAATTC
     5761 CTCGAGGGCT CTTCCTGCTT TGCCAAGGGT ACCAATGTTT TAATGGCGGA TGGGTCTATT
     5821 GAATGTATTG AAAACATTGA GGTTGGTAAT AAGGTCATGG GTAAAGATGG CAGACCTCGT
     5881 GAGGTAATTA AATTGCCCAG AGGAAGAGAA ACTATGTACA GCGTCGTGCA GAAAAGTCAG
     5941 CACAGAGCCC ACAAAAGTGA CTCAAGTCGT GAAGTGCCAG AATTACTCAA GTTTACGTGT
     6001 AATGCGACCC ATGAGTTGGT TGTTAGAACA CCTCGTAGTG TCCGCCGTTT GTCTCGTACC
     6061 ATTAAGGGTG TCGAATATTT TGAAGTTATT ACTTTTGAGA TGGGCCAAAA GAAAGCCCCC
     6121 GACGGTAGAA TTGTTGAGCT TGTCAAGGAA GTTTCAAAGA GCTACCCAAT ATCTGAGGGG
     6181 CCTGAGAGAG CCAACGAATT AGTAGAATCC TATAGAAAGG CTTCAAATAA AGCTTATTTT
     6241 GAGTGGACTA TTGAGGCCAG AGATCTTTCT CTGTTGGGTT CCCATGTTCG TAAAGCTACC
     6301 TACCAGACTT ACGCTCCAAT TCTTTATGAG AATGACCACT TTTTCGACTA CATGCAAAAA
     6361 AGTAAGTTTC ATCTCACCAT TGAAGGTCCA AAAGTACTTG CTTATTTACT TGGTTTATGG
     6421 ATTGGTGATG GATTGTCTGA CAGGGCAACT TTTTCGGTTG ATTCCAGAGA TACTTCTTTG
     6481 ATGGAACGTG TTACTGAATA TGCTGAAAAG TTGAATTTGT GCGCCGAGTA TAAGGACAGA
     6541 AAAGAACCAC AAGTTGCCAA AACTGTTAAT TTGTACTCTA AAGTTGTCAG AGGTAATGGT
     6601 ATTCGCAATA ATCTTAATAC TGAGAATCCA TTATGGGACG CTATTGTTGG CTTAGGATTC
     6661 TTGAAGGACG GTGTCAAAAA TATTCCTTCT TTCTTGTCTA CGGACAATAT CGGTACTCGT
     6721 GAAACATTTC TTGCTGGTCT AATTGATTCT GATGGCTATG TTACTGATGA GCATGGTATT
     6781 AAAGCAACAA TAAAGACAAT TCATACTTCT GTCAGAGATG GTTTGGTTTC CCTTGCTCGT
     6841 TCTTTAGGCT TAGTAGTCTC GGTTAACGCA GAACCTGCTA AGGTTGACAT GAATGTCACC
     6901 AAACATAAAA TTAGTTATGC TATTTATATG TCTGGTGGAG ATGTTTTGCT TAACGTTCTT
     6961 TCGAAGTGTG CCGGCTCTAA AAAATTCAGG CCTGCTCCCG CCGCTGCTTT TGCACGTGAG
     7021 TGCCGCGGAT TTTATTTCGA GTTACAAGAA TTGAAGGAAG ACGATTATTA TGGGATTACT
     7081 TTATCTGATG ATTCTGATCA TCAGTTTTTG CTTGGATCCC AGGTTGTCGT CCATGCATGC
     7141 GGTGGCCTGA CCGGTCTGAA CTCAGGCCTC ACGACAAATC CTGGTGTATC CGCTTGGCAG
     7201 GTCAACACAG CTTATACTGC GGGACAATTG GTCACATATA ACGGCAAGAC GTATAAATGT
     7261 TTGCAGCCCC ACACCTCCTT GGCAGGATGG GAACCATCCA ACGTTCCTGC CTTGTGGCAG
     7321 CTTCAATGAC TGCAGGAAGG GGATCCGGCT GCTAACAAAG CCCGAAAGGA AGCTGAGTTG
     7381 GCTGCTGCCA CCGCTGAGCA ATAACTAGCA TAACCCCTTG GGGCCTCTAA ACGGGTCTTG
     7441 AGGGGTTTTT TGCTGAAAGG AGGAACTATA TCCGGAT
    //

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    图标文献和实验
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    • NEB原核表达系统

      -CN蛋白纯化 系统,可利用还原剂DTT使标签自动裂解,无需外加蛋白酶切割,可以在4度条件下进行,小分子的还原剂也很容易去除;IMPACT-CN蛋白纯化系统包含的表达载体(pTYB载体)有两种,内含肽分别与目的蛋白的C末端(pTYB1和 pTYB2)或N末端(pTYB11和pTYB12)融合.这种在融合蛋白构建方面的机动性极大优化了目的蛋白成功表达和纯化的能力.pTYB2和 pTYB12载体的多克隆位点区设计了相同或者可兼容的酶切位点,使得目的蛋白可选择任一种融合构建载体.而pTYB1及pTYB

    • NEB原核表达系统

      是IMPACT-CN蛋白纯化系统,可利用还原剂DTT使标签自动裂解,无需外加蛋白酶切割,可以在4度条件下进行,小分子的还原剂也很容易去除;IMPACT-CN蛋白纯化系统包含的表达载体(pTYB载体)有两种,内含肽分别与目的蛋白的C末端(pTYB1和 pTYB2)或N末端(pTYB11和pTYB12)融合.这种在融合蛋白构建方面的机动性极大优化了目的蛋白成功表达和纯化的能力.pTYB2和 pTYB12载体的多克隆位点区设计了相同或者可兼容的酶切位点,使得目的蛋白可选择任一种融合构建载体.而pTYB1及pTYB

    • 去Tag不留痕――NEB原核表达系列

      载体(pTYB载体)有两种,内含肽分别与目的蛋白的C末端(pTYB1和pTYB2)或N末端(pTYB11和pTYB12)融合。这种在融合蛋白构建方面的机动性极大优化了目的蛋白成功表达和纯化的能力。pTYB2和pTYB12载体的多克隆位点区设计了相同或者可兼容的酶切位点,使得目的蛋白可选择任一种融合构建载体。而pTYB1及pTYB11载体可让目的基因与内含肽的裂解位点直接相连,在4℃利用 DTT 诱导内含肽的肽键裂解活性,使得最后纯化得到的天然蛋白不带有源自于载体的额外的蛋白残基,同时低温也最大程度地保

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