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pET-44b(+)

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  • 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-44b(+)

    • 库存

      大量

    • 供应商

      钰博生物

    • 规格

      1ug/5ug

    pET-44b(+)

    pET44b载体基本信息

    出品公司: Ybscience
    别名: pET44b, pET 44b
    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: 7310 bp
    5' 测序引物: T7
    5' 测序引物序列:   T7: 5'-TAATACGACTCACTATAGGG-3'
    3' 测序引物: ColiDOWN
    3' 测序引物序列:   ColiDOWN: 5'-TTCACTTCTGAGTTCGGCATG-3'
    载体标签: C-HSV, N-His, C-His, N-Thrombin, N-Nus, N-EK
    载体抗性: Ampicillin
    表达宿主菌: BL21(DE3) 、 Rosetta2(DE3)、C43(DE3)
    备注: Both Nterm and Cterm His tags; Nterm thrombin cleavage site; Nterm 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

    pET44b载体质粒图谱和多克隆位点信息

    pet44b载体图谱
    pet44b多克隆位点

    Feature Name Start End  
    T7_Terminal_primer 69 87
    rrnB_T2_terminator 168 141
    rrnB_T1_terminator 343 300
    rrnB_terminator 466 309
    EK 694 680
    S15 790 746
    6xHIS 2341 2324
    T7_transl_en_RBS 2378 2362
    lacO 2423 2396
    T7_promoter 2441 2423
    tet (300 - 563) 2477 2740
    pBRrevBam_primer 2548 2529
    lacI 2823 3914
    ROP 4486 4677
    pGEX_3_primer 4693 4671
    pBR322_origin 5711 5092
    AmpR_promoter 5798 5826
    Ampicillin 5869 6729
    ORF Start End  
    ORF frame 2 2353 509
    ORF frame 2 803 2131
    ORF frame 2 2324 3082
    ORF frame 3 2955 3914
    ORF frame 1 5869 6729
    Enzyme Name Cut
    PacI 503
    XhoI 530
    PmlI 576
    NotI 592
    EagI 592
    HindIII 599
    KpnI 624
    SalI 626
    PstI 639
    AscI 641
    EcoRI 654
    BamHI 660
    SacI 672
    SmaI 720
    XmaI 718
    SacII 796
    SpeI 827
    BglII 942
    MscI 1074
    StuI 1127
    NcoI 1402
    NdeI 2352
    XbaI 2390
    ApaI 3393
    HpaI 3688
    FspI 6433

     

    pET44b载体简介

    The pET-44 vectors are designed for cloning and high-level expression of peptide sequences fused with the 495 aa Nus•Tag™ protein. Compared to the pET-43.1 series, the pET-44 vectors encode an additional N-terminal His•Tag. 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 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 COLIDOWN primer (Cat. No. 70845-3). Vector encoded sequence can be completely removed when cloning into the PshA I or Sma I sites (as shown below) by cleaving the Nus•Tag fusion protein with enterokinase or thrombin, respectively.

    pET44b载体序列

    LOCUS       pET-44 b(+)    7310 bp     DNA  Circular   SYN
    DEFINITION  pET-44 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..7310
                         /organism="pET-44 b(+)"
                         /mol_type="other DNA"
         misc_feature    69..87
                         /label="T7_Terminal_primer"
         terminator      complement(141..168)
                         /label="rrnB_T2_terminator"
         terminator      complement(300..343)
                         /label="rrnB_T1_terminator"
         terminator      complement(309..466)
                         /label="rrnB_terminator"
         CDS             complement(509..2353)
                         /label="ORF frame 2"
         misc_feature    complement(680..694)
                         /label="EK"     
         misc_feature    complement(746..790)
                         /label="S15"  
         CDS             803..2131
                         /label="ORF frame 2"      
         CDS             2324..3082
                         /label="ORF frame 2"  
         misc_feature    complement(2324..2341)
                         /label="6xHIS"  
         misc_feature    complement(2362..2378)
                         /label="T7_transl_en_RBS" 
         misc_feature    complement(2396..2423)
                         /label="lacO"    
         promoter        complement(2423..2441)
                         /label="T7_promoter"    
         gene            2477..2740
                         /label="tet (300 - 563)"
                         /gene="tet (300 - 563)"
         misc_feature    complement(2529..2548)
                         /label="pBRrevBam_primer"  
         misc_feature    2823..3914
                         /label="lacI" 
         CDS             2955..3914
                         /label="ORF frame 3"   
         misc_feature    4486..4677
                         /label="ROP"  
         misc_feature    complement(4671..4693)
                         /label="pGEX_3_primer"  
         rep_origin      complement(5092..5711)
                         /label="pBR322_origin"  
         promoter        5798..5826
                         /label="AmpR_promoter" 
         gene            5869..6729
                         /label="Ampicillin"
                         /gene="Ampicillin"    
         CDS             5869..6729
                         /label="ORF frame 1"
    ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggata catatttgaa tgtatttaga aaaataaaca
      121 aaagagtttg tagaaacgca aaaaggccat ccgtcaggat ggccttctgc ttaatttgat
      181 gcctggcagt ttatggcggg cgtcctgccc gccaccctcc gggccgttgc ttcgcaacgt
      241 tcaaatccgc tcccggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga
      301 taaaacgaaa ggcccagtct ttcgactgag cctttcgttt tatttgatgc ctggcagttc
      361 cctactctcg catggggaga ccccacacta ccatcggcgc tacggcgttt cacttctgag
      421 ttcggcatgg ggtcaggtgg gaccaccgcg ctactgccgc caggcaaatt ctgttttatc
      481 agcctaggaa cgcccaactt aattaacatt agtggtggtg gtggtggtgc tcgagatcct
      541 ctgggtcttc aggagcgagt tctggctggc ttgcacgtgt atacagctgt gcggccgcaa
      601 gcttcgaacg cgtatcgatg gtaccgtcga cgtcctgcag gcgcgcctgt acagaattcg
      661 gatccacgag ctccggactc ttgtcgtcgt catcaatcgt accagaaccc gcgctgcccc
      721 gggggaccag accagttggc ggtggcgagt ccatgtgctg gcgttcaaat ttcgcagcag
      781 cggtttcttt acccgcggag tgatggtgat ggtgatgacc agaaccacta gtcgcttcgt
      841 caccgaacca gcaaatatta cgggcagcca taatcagtgc tccggctttt tcgtcggtca
      901 acccttcgat atcagccaga tcatcaatgc cctgttcggc gagatcttcc agcgtacaaa
      961 cgccacgggc ggccagtttg aatgccaaat cacgatctac cccttcaagg ttcagcagat
     1021 cgtcagccgg tttgttatca ccgaggcttt cttcctgggc ctgtgcaatg gtggccagtg
     1081 catttttagc acgctcgcgc agtgcttcaa cggtcggctc atcaaggcct tcgatttcca
     1141 acagctcttt catcggcaca taggccaatt cttccagcgt cgagaagcct tcttctacca
     1201 gaacagtcgc gaagtcttcg tcgatgtcga gatatttggt gaaggtgtcg atcgctgcgt
     1261 gcgcttccgc ctgatgctta gcttgcaggt cgtcaacggt catcacgttg agttcccaac
     1321 cgctcagttg cgaagccaga cgcacgttct gaccgttacg gccaatcgcc tgcgccagat
     1381 taccggcttc aacggcgatg tccatggtgt gtttatcttc atccaccacg atagaagcaa
     1441 cgtctgccgg tgccattgcg ttaatcacga actgcgccgg gttatcatcc cacaggacga
     1501 tatcgatacg ctcgccaccc agttcagtag acaccgcctg aacacgcgcg ccacgcatac
     1561 ctacgcaagc acctaccgga tcgatacgtt tatcgttggt tttcaccgcg attttcgcac
     1621 gagaacccgg atcgcgagcc gctgctttaa tttcaatcac ttcttcgccg atttctggca
     1681 cttcaatacg gaacagttcg atcagcattt ccggcttgga acgagtgacg aacagttgcg
     1741 cgccacgcgc ttccgggcga acggaataga gcacgccacg aacgcggtcg ccagggcgga
     1801 agttttcacg cggcagcata tcttcgcgca ggatcacggc ttcagcgttg ttgcccagat
     1861 ccagagagat gttgtcgcgg tttacttttt tcaccacgcc ggtgatgatt tcaccttcgt
     1921 gttcacggaa ctgatcaacc accatcgcac gttcggcttc acgcactttc tgcacgataa
     1981 cctgttttgc cgtctgggta gtgatacggt caaaggtaac agactcaatc tgatcttcaa
     2041 cgtaatcgcc caggttcagg ctttcatctt cataacgtgc ggcttcaagg gtgatttcct
     2101 tggtcggctg ggtgacttca tcaacaacta accagcgacg gaaagtgtca aaatcaccgc
     2161 ttttgcgatc gatctgtacg cggacgtcga tctcttgttc atattttttc tttgttgctg
     2221 tcgccagcgc gctttccaat gcttcgaaaa tcttctcgcg aggtagcgcc ttttcattgg
     2281 atacggcttc aactacagcc aaaatttctt tgttcatact agaatggtga tggtggtgat
     2341 ggctgctgcc catatgtata tctccttctt aaagttaaac aaaattattt ctagagggga
     2401 attgttatcc gctcacaatt cccctatagt gagtcgtatt aatttcgcgg gatcgagatc
     2461 gatctcgatc ctctacgccg gacgcatcgt ggccggcatc accggcgcca caggtgcggt
     2521 tgctggcgcc tatatcgccg acatcaccga tggggaagat cgggctcgcc acttcgggct
     2581 catgagcgct tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg gactgttggg
     2641 cgccatctcc ttgcatgcac cattccttgc ggcggcggtg ctcaacggcc tcaacctact
     2701 actgggctgc ttcctaatgc aggagtcgca taagggagag cgtcgagatc ccggacacca
     2761 tcgaatggcg caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca
     2821 gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt
     2881 atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa
     2941 aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg
     3001 cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt
     3061 cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt
     3121 cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc
     3181 aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg
     3241 aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca
     3301 acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat ctggtcgcat
     3361 tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc
     3421 gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata gcggaacggg
     3481 aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg aatgagggca
     3541 tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca atgcgcgcca
     3601 ttaccgagtc cgggctgcgc gttggtgcgg acatctcggt agtgggatac gacgataccg
     3661 aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg
     3721 ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc
     3781 agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg
     3841 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     3901 aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc tcactcatta ggcaccggga
     3961 tctcgaccga tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc
     4021 atgactagca tgatcgtgct cctgtcgttg aggacccggc taggctggcg gggttgcctt
     4081 actggttagc agaatgaatc accgatacgc gagcgaacgt gaagcgactg ctgctgcaaa
     4141 acgtctgcga cctgagcaac aacatgaatg gtcttcggtt tccgtgtttc gtaaagtctg
     4201 gaaacgcgga agtcagcgcc ctgcaccatt atgttccgga tctgcatcgc aggatgctgc
     4261 tggctaccct gtggaacacc tacatctgta ttaacgaagc gctggcattg accctgagtg
     4321 atttttctct ggtcccgccg catccatacc gccagttgtt taccctcaca acgttccagt
     4381 aaccgggcat gttcatcatc agtaacccgt atcgtgagca tcctctctcg tttcatcggt
     4441 atcattaccc ccatgaacag aaatccccct tacacggagg catcagtgac caaacaggaa
     4501 aaaaccgccc ttaacatggc ccgctttatc agaagccaga cattaacgct tctggagaaa
     4561 ctcaacgagc tggacgcgga tgaacaggca gacatctgtg aatcgcttca cgaccacgct
     4621 gatgagcttt accgcagctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     4681 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     4741 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt
     4801 agcgatagcg gagtgtatac tggcttaact atgcggcatc agagcagatt gtactgagag
     4861 tgcaccatat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     4921 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4981 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5041 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     5101 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5161 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5221 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5281 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5341 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     5401 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5461 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5521 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     5581 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5641 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5701 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5761 tttggtcatg acctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     5821 gacaataacc ctgataaatg cttcaatcat gattgaaaaa ggaagagtat gagtattcaa
     5881 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     5941 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     6001 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     6061 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     6121 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     6181 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     6241 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     6301 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     6361 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     6421 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     6481 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     6541 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     6601 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     6661 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     6721 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     6781 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgcc atcgtgccgc
     6841 cagaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     6901 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     6961 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     7021 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     7081 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     7141 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     7201 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     7261 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
    //

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