pET-31b(+)载体

pET-31b(+)载体(Cat. No. 69952-3)是设计用来高水平表达蛋白质的载体，载体融合表达有125个氨基酸的酮类固醇异构酶(ketosteroid isomerase protein) 。载体上面的AlwN I单克隆位点允许单向插入串联重复的通过蛋氨酸（methionine）分隔开的多肽序列。

pET31b载体的单一的多克隆位点见上面的环状质粒图谱。注意：载体序列是以pBR322质粒的编码规矩进行编码的，所以T7蛋白表达区在质粒图谱上面是反向的。

T7 RNA聚合酶启动的克隆和表达区域在质粒图谱中也被标注了出来。质粒的F1复制子是被定向的，所以在T7噬菌体聚合酶的作用下，包含有蛋白编码序列的病毒 粒子能够产生，并启动蛋白表达，同时蛋白表达将被T7终止子序列(Cat. No. 69337-3)的作用下终止蛋白翻译。

LOCUS       pET-31b(+)    5742 bp     DNA  Circular    SYNDEFINITION  pET-31b(+)ACCESSION   KEYWORDS    SOURCE        ORGANISM  other sequences; artificial sequences; vectors.COMMENT     This file is created by Vector NTI            http://www.biofeng.com/COMMENT     ORIGDB|GenBankCOMMENT     VNTAUTHORNAME|biofeng.com|COMMENT     VNTOAUTHORAD1|Novagen Inc.|FEATURES             Location/Qualifiers     source          1..5742                     /organism="pET-31b(+)"                     /mol_type="other DNA"     terminator      complement(1..129)                     /label="T7_terminator"     misc_feature    69..87                     /label="T7_Terminal_primer"     misc_feature    complement(553..569)                     /label="T7_transl_en_RBS"     misc_feature    complement(587..614)                     /label="lacO"     promoter        complement(614..632)                     /label="T7_promoter"     gene            664..927                     /label="tet (300 - 563)"                     /gene="tet (300 - 563)"     misc_feature    complement(716..735)                     /label="pBRrevBam_primer"     CDS             769..1269                     /label="ORF frame 1"     misc_feature    1010..2101                     /label="lacI"     CDS             1142..2101                     /label="ORF frame 2"     gene            2160..2435                     /label="tet (576 - 851)"                     /gene="tet (576 - 851)"     misc_feature    2910..3101                     /label="ROP"     misc_feature    complement(3095..3117)                     /label="pGEX_3_primer"     gene            complement(4284..5144)                     /label="Ampicillin"                     /gene="Ampicillin"     CDS             complement(4284..5144)                     /label="ORF frame 2"     promoter        complement(5186..5214)                     /label="AmpR_promoter"ORIGIN    1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa   61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt  121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagcagcatc tggcatgcgt  181 gaatattctt ctcgccaaac aaggcgcgga tgctcaccac cttgccggcg ccattgaagc  241 gaaagtgatc gatgggcgca actacggtct tgcggccctg atactcgaag ctgacggtga  301 aagcgaaggc cgcttcgttg gcgaccgcgc gtacctcctg cgtcagctcc accgccaaag  361 gcagtttgag cgagttggcg taaaactcac gaatcgcagc cgtaccggac ctgggctcgg  421 aacccacggg gtcttccacc gtggcgtcat cggcaaacag cgcgacgatg ccgtccagat  481 cgccggcatt gagcgcagcc acaaagcgct gtaccacggc ggtgatgtgt tctggggtat  541 gcatatgtat atctccttct taaagttaaa caaaattatt tctagagggg aattgttatc  601 cgctcacaat tcccctatag tgagtcgtat taatttcgcg ggatcgagat ctcgatcctc  661 tacgccggac gcatcgtggc cggcatcacc ggcgccacag gtgcggttgc tggcgcctat  721 atcgccgaca tcaccgatgg ggaagatcgg gctcgccact tcgggctcat gagcgcttgt  781 ttcggcgtgg gtatggtggc aggccccgtg gccgggggac tgttgggcgc catctccttg  841 catgcaccat tccttgcggc ggcggtgctc aacggcctca acctactact gggctgcttc  901 ctaatgcagg agtcgcataa gggagagcgt cgagatcccg gacaccatcg aatggcgcaa  961 aacctttcgc ggtatggcat gatagcgccc ggaagagagt caattcaggg tggtgaatgt 1021 gaaaccagta acgttatacg atgtcgcaga gtatgccggt gtctcttatc agaccgtttc 1081 ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc 1141 gatggcggag ctgaattaca ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc 1201 gttgctgatt ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc 1261 ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga tggtagaacg 1321 aagcggcgtc gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg 1381 gctgatcatt aactatccgc tggatgacca ggatgccatt gctgtggaag ctgcctgcac 1441 taatgttccg gcgttatttc ttgatgtctc tgaccagaca cccatcaaca gtattatttt 1501 ctcccatgaa gacggtacgc gactgggcgt ggagcatctg gtcgcattgg gtcaccagca 1561 aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc tggctggctg 1621 gcataaatat ctcactcgca atcaaattca gccgatagcg gaacgggaag gcgactggag 1681 tgccatgtcc ggttttcaac aaaccatgca aatgctgaat gagggcatcg ttcccactgc 1741 gatgctggtt gccaacgatc agatggcgct gggcgcaatg cgcgccatta ccgagtccgg 1801 gctgcgcgtt ggtgcggata tctcggtagt gggatacgac gataccgaag acagctcatg 1861 ttatatcccg ccgttaacca ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt 1921 ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt 1981 ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg caaaccgcct ctccccgcgc 2041 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg 2101 agcgcaacgc aattaatgta agttagctca ctcattaggc accgggatct cgaccgatgc 2161 ccttgagagc cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg 2221 ccgcacttat gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct 2281 gggtcatttt cggcgaggac cgctttcgct ggagcgcgac gatgatcggc ctgtcgcttg 2341 cggtattcgg aatcttgcac gccctcgctc aagccttcgt cactggtccc gccaccaaac 2401 gtttcggcga gaagcaggcc attatcgccg gcatggcggc cccacgggtg cgcatgatcg 2461 tgctcctgtc gttgaggacc cggctaggct ggcggggttg ccttactggt tagcagaatg 2521 aatcaccgat acgcgagcga acgtgaagcg actgctgctg caaaacgtct gcgacctgag 2581 caacaacatg aatggtcttc ggtttccgtg tttcgtaaag tctggaaacg cggaagtcag 2641 cgccctgcac cattatgttc cggatctgca tcgcaggatg ctgctggcta ccctgtggaa 2701 cacctacatc tgtattaacg aagcgctggc attgaccctg agtgattttt ctctggtccc 2761 gccgcatcca taccgccagt tgtttaccct cacaacgttc cagtaaccgg gcatgttcat 2821 catcagtaac ccgtatcgtg agcatcctct ctcgtttcat cggtatcatt acccccatga 2881 acagaaatcc cccttacacg gaggcatcag tgaccaaaca ggaaaaaacc gcccttaaca 2941 tggcccgctt tatcagaagc cagacattaa cgcttctgga gaaactcaac gagctggacg 3001 cggatgaaca ggcagacatc tgtgaatcgc ttcacgacca cgctgatgag ctttaccgca 3061 gctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga 3121 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag 3181 cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt 3241 atactggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatatgcgg 3301 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc 3361 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca 3421 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca 3481 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg 3541 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg 3601 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt 3661 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt 3721 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc 3781 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt 3841 gagtccaacc cggtaagaca cgacttatcg ccactggcag ctggtaacag gattagcaga 3901 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact 3961 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt 4021 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag 4081 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg 4141 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa 4201 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata 4261 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg 4321 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata 4381 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg 4441 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct 4501 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt 4561 tcgccagtta atagtttgcg caacgttgtt gccattgctg caggcatcgt ggtgtcacgc 4621 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga 4681 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt 4741 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc 4801 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa 4861 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca 4921 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca 4981 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct 5041 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc 5101 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa 5161 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt 5221 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgaaatt 5281 gtaaacgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt 5341 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg 5401 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc 5461 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca 5521 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga 5581 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa 5641 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc 5701 gccgcgctta atgcgccgct acagggcgcg tcccattcgc ca//