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pET-42c

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产物名称: pET-42c
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产物展商: XYbscience
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简单介绍

pET-42c菌株发货前均经过严格的多重验证，如存在质量问题，请在收到产物的三个月内通知我司。pET-42c收到质粒后请短暂离心，加入20μ濒 ddH2O溶解质粒，取2μl转化至对应感受态中，挑取单克隆重新提取质粒后使用。


pET-42c  的详细介绍
 

pET-42c

  • 产物信息
  • 基本信息
  • 质粒介绍
  • 质粒图谱
  • 质粒序列

产物信息

产物货号 产物名称 产物规格 优惠价
XY0373 pET-42c

20μ濒

?请询价

使用说明

信裕质粒平台的各批次质粒菌株发货前均经过严格的多重验证，如存在质量问题，请在收到产物的三个月内通知我司。收到质粒后请短暂离心，加入20μ濒 ddH2O溶解质粒，取2μl转化至对应感受态中，挑取单克隆重新提取质粒后使用。如需获取其他详细信息请登录我司官网查询。

基本信息

启动子: T7/lac
复制子: ColE1 ori，F1 ori
终止子: T7 terminator
质粒分类: 大肠杆菌载体；笔贰罢系列表达质粒
质粒大小: 5931bp
质粒标签: N-GST, N-6×His, N-S, C-6×His, N-Thrombin, N-Factor Xa
原核抗性: 卡那霉素碍补苍
克隆菌株: 顿贬5α
培养条件: 37℃，有氧，尝叠
表达宿主: BL21(DE3)
培养条件: 37℃，有氧，尝叠
诱导方式: 滨笔罢骋或乳糖及其类似物
5'测序引物: T7:TAATACGACTCACTATAGGG
3'测序引物: T7-ter:TGCTAGTTATTGCTCAGCGG

质粒介绍

       pET42系列载体是用来进行克隆和高水平表达含有220个氨基酸的GST融合纯化标签的蛋白表达载体。
       pET42a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终止子序列的作用下终止蛋白翻译。
把表达的基因插入到载体的PshAI限制性内切酶位点时，可以通过Factor Xa蛋白酶将融合表达的GST标签完全切除。

质粒图谱

质粒序列

LOCUS       Exported                5931 bp ds-DNA     circular SYN 06-SEP-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pET-42c
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5931)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, September 6, 2016 from SnapGene Viewer 3.1.4
            http://www.miaolingbio.com
FEATURES             Location/Qualifiers
     source          1..5931
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     terminator      26..73
                     /note="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
     CDS             complement(150..173)
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /note="8xHis"
                     /translation="HHHHHHHH"
     CDS             complement(266..277)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /note="Factor Xa site"
                     /translation="IEGR"
     CDS             complement(308..352)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic
                     ribonuclease A"
                     /note="S-Tag"
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(368..385)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /note="thrombin site"
                     /translation="LVPRGS"
     CDS             complement(395..412)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     CDS             complement(440..1093)
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma
                     japonicum"
                     /note="GST"
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     protein_bind    1138..1162
                     /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)."
     promoter        complement(1163..1181)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1494..1571
                     /gene="lacI"
                     /note="lacI promoter"
     CDS             1572..2654
                     /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"
     CDS             3226..3417
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein"
                     /note="rop"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     rep_origin      complement(3847..4435)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number colE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             4557..5372
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /note="KanR"
                     /note="confers resistance to kanamycin"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      complement(5465..5920)
                     /direction=LEFT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgtttagcag cctaggtatt aatcaattag tggtggtggt ggtggtggtg gtgctcgagt
      181 gcggccgcaa gcttgtcgac ggagctcgcc tgcaggcgcg ccaaggcctg tacagaattc
      241 ggatccccga tatcgccatg gacccgcgtc cctcaatacc ggagccacca ccggtaccca
      301 gatctgggct gtccatgtgc tggcgttcga atttagcagc agcggtttct ttcataccaa
      361 ttgcagtact accgcgtggc accagacccg cggagtgatg gtgatggtga tgaccagaac
      421 cactagttga accatccgat tttggaggat ggtcgccacc accaaacgtg gcttgccagc
      481 cctgcaaagg ccatgctata tacttgctgg atttcaagta cttatcaatt tgtgggatag
      541 cttcaatacg ttttttaaaa caaactaatt ttgggaacgc atccaggcac attgggtcca
      601 tgtataaaac aacatcaaga gcgtcataca acatgaagtc aggatgggtt acatgatcac
      661 catttaaata tgttttatga cataaacgat cttcgaacat tttcagcatt tcaggtagct
      721 tgctaagaaa atcaactttg agagtttcaa agtctttact atatgcaatt ctcgaaacac
      781 cgtatctaat atccaaaacc gctccttcaa gcattgaaat ctctgcacgc tcttttggac
      841 aaccacccaa catgttgtgc ttgtcagcta tataacgtat gatggccata gactgtgtta
      901 atttaacatc accatcaata taataaggaa gattgggaaa ctccaaaccc aattcaaact
      961 ttttgtttcg ccatttatca ccttcatcgc gctcatacaa atgctcttca tatttttctt
     1021 caagatattc caaaagaagt cgagtgggtt gcacaaggcc cttaattttc caataaccta
     1081 gtatagggga catatgtata tctccttctt aaagttaaac aaaattattt ctagagggga
     1141 attgttatcc gctcacaatt cccctatagt gagtcgtatt aatttcgcgg gatcgagatc
     1201 gatctcgatc ctctacgccg gacgcatcgt ggccggcatc accggcgcca caggtgcggt
     1261 tgctggcgcc tatatcgccg acatcaccga tggggaagat cgggctcgcc acttcgggct
     1321 catgagcgct tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg gactgttggg
     1381 cgccatctcc ttgcatgcac cattccttgc ggcggcggtg ctcaacggcc tcaacctact
     1441 actgggctgc ttcctaatgc aggagtcgca taagggagag cgtcgagatc ccggacacca
     1501 tcgaatggcg caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca
     1561 gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt
     1621 atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa
     1681 aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg
     1741 cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt
     1801 cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt
     1861 cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc
     1921 aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg
     1981 aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca
     2041 acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat ctggtcgcat
     2101 tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc
     2161 gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata gcggaacggg
     2221 aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg aatgagggca
     2281 tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca atgcgcgcca
     2341 ttaccgagtc cgggctgcgc gttggtgcgg acatctcggt agtgggatac gacgataccg
     2401 aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg
     2461 ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc
     2521 agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg
     2581 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     2641 aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc tcactcatta ggcaccggga
     2701 tctcgaccga tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc
     2761 atgactagca tgatcgtgct cctgtcgttg aggacccggc taggctggcg gggttgcctt
     2821 actggttagc agaatgaatc accgatacgc gagcgaacgt gaagcgactg ctgctgcaaa
     2881 acgtctgcga cctgagcaac aacatgaatg gtcttcggtt tccgtgtttc gtaaagtctg
     2941 gaaacgcgga agtcagcgcc ctgcaccatt atgttccgga tctgcatcgc aggatgctgc
     3001 tggctaccct gtggaacacc tacatctgta ttaacgaagc gctggcattg accctgagtg
     3061 atttttctct ggtcccgccg catccatacc gccagttgtt taccctcaca acgttccagt
     3121 aaccgggcat gttcatcatc agtaacccgt atcgtgagca tcctctctcg tttcatcggt
     3181 atcattaccc ccatgaacag aaatccccct tacacggagg catcagtgac caaacaggaa
     3241 aaaaccgccc ttaacatggc ccgctttatc agaagccaga cattaacgct tctggagaaa
     3301 ctcaacgagc tggacgcgga tgaacaggca gacatctgtg aatcgcttca cgaccacgct
     3361 gatgagcttt accgcagctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     3421 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     3481 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt
     3541 agcgatagcg gagtgtatac tggcttaact atgcggcatc agagcagatt gtactgagag
     3601 tgcaccatat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     3661 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     3721 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     3781 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3841 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3901 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3961 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4021 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4081 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4141 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4201 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4261 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     4321 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4381 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4441 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4501 tttggtcatg aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga
     4561 gccatattca acgggaaacg tcttgctcta ggccgcgatt aaattccaac atggatgctg
     4621 atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc
     4681 gattgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg
     4741 ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc
     4801 cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc
     4861 ccgggaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg
     4921 atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta
     4981 acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg
     5041 atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa
     5101 tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg
     5161 ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa
     5221 tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt
     5281 cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc
     5341 agtttcattt gatgctcgat gagtttttct aagaattaat tcatgagcgg atacatattt
     5401 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     5461 cctgaaattg taaacgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     5521 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     5581 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     5641 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     5701 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     5761 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     5821 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     5881 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc a
//
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