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pGEX-KG Plasmid

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    • 文献和实验
    • 技术资料
    • 保存条件

      负20摄氏度

    • 保质期

      3个月

    • 英文名

      pGEX-KG

    • 库存

      50

    • 供应商

      LSMBIO

    • 规格

      2ug

    pGEX-KG

    Search name
    pGEX-KG,Plasmid pGEX-KG,pGEX-KG vector

     

    pGEX-KG Plasmid Information

    Promoter: Tac/lac

    Replicon: ColE1 ori

    Plasmid classification: Escherichia coli vector; PGEX series expression plasmid

    Plasmid size: 5005bp

    Plasmid label: N-GST, N-thrombin

    Prokaryotic resistance: ampicillin Amp

    Cloned strain: DH5 alpha

    Culture conditions: 37 centigrade, aerobic, LB

    Expression host: Escherichia coli

    Culture conditions: 37 centigrade, aerobic, LB

    Inducement: IPTG or lactose and its analogues

    5'sequencing primers: pGEX5: GGGCTGGCAAGCCACGTTTGGTG

    3'sequencing primers: pGEX3: CCGGGAGCTGCATGTGTCAGAGG

    Note: the recombinant protein can be purified by GST affinity column

     

    pGEX-KG Plasmid Description

    The size of the pGEX-KG carrier is 5005bp, which can grow on the Amp plate, and the GST label can be used to purify the recombinant protein with the GST affinity column

    pGEX-KG-site

     

    pGEX-KG Plasmid Sequence

    LOCUS       Exported File           5005 bp ds-DNA    circular SYN 29-1-2015
    DEFINITION  .
    ACCESSION   .
    VERSION     .
    KEYWORDS    pGEX-KG
    SOURCE      synthetic DNA construct
      ORGANISM  synthetic DNA construct
    REFERENCE   1  (bases 1 to 5005)
      AUTHORS   .
      TITLE     Direct Submission
      JOURNAL   Exported 2015-1-29 
               
    FEATURES             Location/Qualifiers
         source          1..5005
                         /organism="synthetic DNA construct"
                         /mol_type="other DNA"
         misc_feature    183..211
                         /note="tac promoter"
                         /note="strong E. coli promoter; hybrid between the trp and
                         lac?UV5 promoters"
         protein_bind    219..235
                         /bound_moiety="lac repressor encoded by lacI"
                         /note="lac operator"
                         /note="The lac repressor binds to the lac operator to
                         inhibit transcription in E. coli. This inhibition can be
                         relieved by adding lactose or
                         isopropyl-beta-D-thiogalactopyranoside (IPTG)."
         CDS             258..911
                         /codon_start=1
                         /gene="
                         "
                         /product="glutathione S-transferase from Schistosoma
                         japonicum"
                         /note="GST"
                         /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                         FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                         GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                         YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
         CDS             918..935
                         /codon_start=1
                         /product="thrombin recognition and cleavage site"
                         /note="thrombin site"
                         /translation="LVPRGS"
         promoter        1308..1412
                         /gene="bla"
                         /note="AmpR promoter"
         CDS             1413..2273
                         /codon_start=1
                         /gene="bla"
                         /product="beta-lactamase"
                         /note="AmpR"
                         /note="confers resistance to ampicillin, carbenicillin, and
                         related antibiotics"
                         /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                         ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                         PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                         EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                         LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                         LIKHW"
         rep_origin      2444..3032
                         /direction=RIGHT
                         /note="ori"
                         /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                         replication"
         promoter        3276..3353
                         /gene="lacI (mutant)"
                         /note="lacIq promoter"
                         /note="In the lacIq allele, a single base change in the
                         promoter boosts expression of the lacI gene about 10-fold."
         CDS             3354..4436
                         /codon_start=1
                         /gene="lacI"
                         /product="lac repressor"
                         /note="lacI"
                         /note="The lac repressor binds to the lac operator to
                         inhibit transcription in E. coli. This inhibition can be
                         relieved by adding lactose or
                         isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                         /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                         NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                         EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                         EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                         MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                         YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                         ALADSLMQLARQVSRLESGQ"
         protein_bind    4449..4470
                         /bound_moiety="E. coli catabolite activator protein"
                         /note="CAP binding site"
                         /note="CAP binding activates transcription in the presence
                         of cAMP."
         promoter        4485..4515
                         /note="lac promoter"
                         /note="promoter for the E. coli lac operon"
         protein_bind    4523..4539
                         /bound_moiety="lac repressor encoded by lacI"
                         /note="lac operator"
                         /note="The lac repressor binds to the lac operator to
                         inhibit transcription in E. coli. This inhibition can be
                         relieved by adding lactose or
                         isopropyl-beta-D-thiogalactopyranoside (IPTG)."
         primer_bind     4547..4563
                         /note="M13 rev"
                         /note="common sequencing primer, one of multiple similar
                         variants"
         primer_bind     complement(4579..4595)
                         /note="M13 fwd"
                         /note="common sequencing primer, one of multiple similar
                         variants"
    ORIGIN
            1 acgttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
           61 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
          121 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
          181 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
          241 cacaggaaac agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc
          301 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc
          361 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc
          421 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata
          481 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc
          541 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact
          601 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag
          661 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt
          721 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa
          781 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat
          841 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc
          901 atcctccaaa atcggatctg gttccgcgtg gatccccggg aatttccggt ggtggtggtg
          961 gaattctaga ctccatgggt cgactcgagc tcaagcttat tcatcgtgac tgactgacga
         1021 tctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
         1081 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
         1141 cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt
         1201 ataattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
         1261 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
         1321 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
         1381 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
         1441 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
         1501 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
         1561 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
         1621 ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg
         1681 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
         1741 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
         1801 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
         1861 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
         1921 ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca
         1981 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
         2041 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
         2101 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
         2161 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
         2221 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
         2281 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
         2341 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
         2401 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
         2461 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
         2521 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
         2581 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
         2641 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
         2701 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
         2761 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
         2821 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
         2881 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
         2941 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
         3001 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
         3061 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
         3121 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
         3181 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
         3241 cgcatctgtg cggtatttca caccgcataa attccgacac catcgaatgg tgcaaaacct
         3301 ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac
         3361 cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg
         3421 tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg
         3481 cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc
         3541 tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga
         3601 ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg
         3661 gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga
         3721 tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg
         3781 ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt attttctccc
         3841 atgaagacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg
         3901 cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata
         3961 aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca
         4021 tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc
         4081 tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc
         4141 gcgttggtgc ggatatctcg gtagtgggat acgacgatac cgaagacagc tcatgttata
         4201 tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc
         4261 gcttgctgca actctctcag ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac
         4321 tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg
         4381 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
         4441 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
         4501 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
         4561 gaccatgatt acggattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
         4621 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
         4681 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgctt
         4741 tgcctggttt ccggcaccag aagcggtgcc ggaaagctgg ctggagtgcg atcttcctga
         4801 ggccgatact gtcgtcgtcc cctcaaactg gcagatgcac ggttacgatg cgcccatcta
         4861 caccaacgta acctatccca ttacggtcaa tccgccgttt gttcccacgg agaatccgac
         4921 gggttgttac tcgctcacat ttaatgttga tgaaagctgg ctacaggaag gccagacgcg
         4981 aattattttt gatggcgttg gaatt
    //

     

     

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