pET-42a

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
XY0476	pET-42a	20μ濒	?请加蚕蚕询价

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/laco promoter
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:	5930bp
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:	大肠杆菌顿贬5α
培养条件:	37℃，有氧，尝叠
表达宿主:	大肠杆菌叠尝21(顿贰3)
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                5930 bp ds-DNA     circular SYN 10-8-2015
DEFINITION synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Untitled 3
SOURCE      synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE   1 (bases 1 to 5930)
AUTHORS   .
TITLE     Direct Submission
JOURNAL   Exported 2015-8-10 from MLCC
            http://www.miaoningbio.com
FEATURES             Location/Qualifiers
     source          1..5930
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1100..1122
                     /organism="Enterobacteria phage T7"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:10760"
     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(265..276)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /note="Factor Xa site"
                     /translation="IEGR"
     CDS             complement(307..351)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic
                     ribonuclease A"
                     /note="S-Tag"
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(367..384)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /note="thrombin site"
                     /translation="LVPRGS"
     CDS             complement(394..411)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     CDS             complement(439..1092)
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma
                     japonicum"
                     /note="GST"
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     RBS             1100..1122
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1137..1161
                     /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(1162..1180)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1493..1570
                     /gene="lacI"
                     /note="lacI promoter"
     CDS             1571..2653
                     /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             3225..3416
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /note="rop"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    3518..3660
                     /note="bom"
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(3846..4434)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             4556..5371
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /note="KanR"
                     /note="confers resistance to kanamycin in bacteria or G418
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      complement(5464..5919)
                     /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 tatcccatgg acccgcgtcc ctcaataccg gagccaccac cggtacccag
      301 atctgggctg tccatgtgct ggcgttcgaa tttagcagca gcggtttctt tcataccaat
      361 tgcagtacta ccgcgtggca ccagacccgc ggagtgatgg tgatggtgat gaccagaacc
      421 actagttgaa ccatccgatt ttggaggatg gtcgccacca ccaaacgtgg cttgccagcc
      481 ctgcaaaggc catgctatat acttgctgga tttcaagtac ttatcaattt gtgggatagc
      541 ttcaatacgt tttttaaaac aaactaattt tgggaacgca tccaggcaca ttgggtccat
      601 gtataaaaca acatcaagag cgtcatacaa catgaagtca ggatgggtta catgatcacc
      661 atttaaatat gttttatgac ataaacgatc ttcgaacatt ttcagcattt caggtagctt
      721 gctaagaaaa tcaactttga gagtttcaaa gtctttacta tatgcaattc tcgaaacacc
      781 gtatctaata tccaaaaccg ctccttcaag cattgaaatc tctgcacgct cttttggaca
      841 accacccaac atgttgtgct tgtcagctat ataacgtatg atggccatag actgtgttaa
      901 tttaacatca ccatcaatat aataaggaag attgggaaac tccaaaccca attcaaactt
      961 tttgtttcgc catttatcac cttcatcgcg ctcatacaaa tgctcttcat atttttcttc
     1021 aagatattcc aaaagaagtc gagtgggttg cacaaggccc ttaattttcc aataacctag
     1081 tataggggac atatgtatat ctccttctta aagttaaaca aaattatttc tagaggggaa
     1141 ttgttatccg ctcacaattc ccctatagtg agtcgtatta atttcgcggg atcgagatcg
     1201 atctcgatcc tctacgccgg acgcatcgtg gccggcatca ccggcgccac aggtgcggtt
     1261 gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca cttcgggctc
     1321 atgagcgctt gtttcggcgt gggtatggtg gcaggccccg tggccggggg actgttgggc
     1381 gccatctcct tgcatgcacc attccttgcg gcggcggtgc tcaacggcct caacctacta
     1441 ctgggctgct tcctaatgca ggagtcgcat aagggagagc gtcgagatcc cggacaccat
     1501 cgaatggcgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag
     1561 ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta
     1621 tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa
     1681 agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc
     1741 gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc
     1801 gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc
     1861 gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca
     1921 acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga
     1981 agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa
     2041 cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt
     2101 gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg
     2161 tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga
     2221 aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat
     2281 cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat
     2341 taccgagtcc gggctgcgcg ttggtgcgga catctcggta gtgggatacg acgataccga
     2401 agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg
     2461 gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca
     2521 gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc
     2581 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     2641 aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag gcaccgggat
     2701 ctcgaccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca
     2761 tgactagcat gatcgtgctc ctgtcgttga ggacccggct aggctggcgg ggttgcctta
     2821 ctggttagca gaatgaatca ccgatacgcg agcgaacgtg aagcgactgc tgctgcaaaa
     2881 cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt ccgtgtttcg taaagtctgg
     2941 aaacgcggaa gtcagcgccc tgcaccatta tgttccggat ctgcatcgca ggatgctgct
     3001 ggctaccctg tggaacacct acatctgtat taacgaagcg ctggcattga ccctgagtga
     3061 tttttctctg gtcccgccgc atccataccg ccagttgttt accctcacaa cgttccagta
     3121 accgggcatg ttcatcatca gtaacccgta tcgtgagcat cctctctcgt ttcatcggta
     3181 tcattacccc catgaacaga aatccccctt acacggaggc atcagtgacc aaacaggaaa
     3241 aaaccgccct taacatggcc cgctttatca gaagccagac attaacgctt ctggagaaac
     3301 tcaacgagct ggacgcggat gaacaggcag acatctgtga atcgcttcac gaccacgctg
     3361 atgagcttta ccgcagctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     3421 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     3481 gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta
     3541 gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg tactgagagt
     3601 gcaccatata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     3661 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     3721 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     3781 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     3841 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     3901 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     3961 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4021 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4081 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4141 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4201 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4261 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     4321 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4381 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     4441 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     4501 ttggtcatga acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag
     4561 ccatattcaa cgggaaacgt cttgctctag gccgcgatta aattccaaca tggatgctga
     4621 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
     4681 attgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
     4741 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc
     4801 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc
     4861 cgggaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
     4921 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
     4981 cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga
     5041 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
     5101 gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
     5161 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
     5221 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
     5281 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
     5341 gtttcatttg atgctcgatg agtttttcta agaattaatt catgagcgga tacatatttg
     5401 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     5461 ctgaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     5521 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     5581 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     5641 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     5701 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     5761 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     5821 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     5881 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
//

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