pTrcHisB

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
XY0908	pTrcHisB	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:	Trc/lac
复制子:	PBR322 ori
终止子:	rrnB TT
质粒分类:	大肠杆菌载体；辫罢谤肠系列表达质粒
质粒大小:	4404bp
质粒标签:	狈-贬颈蝉，狈-贰碍
原核抗性:	氨苄青霉素础尘辫
克隆菌株:	顿贬5α
培养条件:	37℃，有氧，尝叠
表达宿主:	大肠杆菌
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:	pTrcHis-F: GAGGTATATATTAATGTATCG
3'测序引物:	pTrcHis-R:GATTTAATCTGTATCAGG
备注:	辫罢谤肠表达质粒

质粒介绍

辫罢谤肠贬颈蝉载体是来由辫叠搁322质粒衍生而来的表达载体,设计用来在大肠杆菌中进行重组蛋白的高效表达和纯化。
载体上的罢谤肠启动子和谤谤苍叠抗终止区使得高水平蛋白表达成为可能.罢谤肠启动子由罢谤辫启动子的-35区域和尝补肠启动子的-10区域共同组成。辫罢谤肠贬颈蝉载体同时包含有一个尝补肠滨辩基因，该基因编码了尝补肠启动子的抑制蛋白。尝补肠滨辩基因的存在使得宿主菌是尝补肠滨辩+还是尝补肠滨辩-都可以高效抑制插入基因的转录。
进行蛋白表达时，大肠杆菌需要生长到对数生长期，然后加入1mM IPTG，去除阻遏蛋白，诱导表达。载体上的迷你顺反子结构能够增强插入基因的转录水平，从而高水平表达目的蛋白。DNA插入片段位于载体表达片段下游，在其上游有一个N-端融合表达多肽序列，该序列包含有ATG起始密码子，6xHis标签，T7噬菌体10号基因的翻译增强子，X press抗原表位，肠激酶酶切识别位点。

质粒图谱

质粒序列

LOCUS       Exported                4404 bp ds-DNA     circular SYN 29-七月-2016
DEFINITION synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pTrcHisB
SOURCE      synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE   1 (bases 1 to 4404)
AUTHORS   .
TITLE     Direct Submission
JOURNAL   Exported 2016年7月29日 from SnapGene Viewer 2.8.1
            http://www.miaolingbio.com
FEATURES             Location/Qualifiers
     source          1..4404
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    193..222
                     /note="trc promoter"
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    230..246
                     /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)."
     misc_feature    263..332
                     /note="rrnG antiterminator"
                     /note="antiterminator from the E. coli rrnG leader region
                     (Berg et al., 1989)"
     CDS             383..409
                     /codon_start=1
                     /product="synthetic cistron containing a ribosome binding
                     site (Shine-Dalgarno sequence), for enhancing the bacterial
                     expression of a downstream cistron (Schoner, 1997)"
                     /note="minicistron"
                     /note="This first cistron should terminate 3 bp upstream of
                     the ATG for the second cistron."
                     /translation="MYRLNKEE"
     CDS             425..442
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     CDS             446..478
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /note="T7 tag (gene 10 leader)"
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             482..505
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /note="Xpress(TM) tag"
                     /translation="DLYDDDDK"
     misc_feature    505..554
                     /note="MCS"
     terminator      757..843
                     /gene="Escherichia coli rrnB"
                     /note="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      935..962
                     /note="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        982..1073
                     /gene="bla"
                     /note="AmpR promoter"
     CDS             1074..1934
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2105..2693
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     misc_feature    2879..3019
                     /note="bom"
                     /note="basis of mobility region from pBR322"
     promoter        3205..3282
                     /gene="lacI"
                     /note="lacI promoter"
     CDS             3283..4365
                     /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
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
ORIGIN
        1 gtttgacagc ttatcatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc
       61 ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc
      121 gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc
      181 tgaaatgagc tgttgacaat taatcatccg gctcgtataa tgtgtggaat tgtgagcgga
      241 taacaatttc acacaggaaa cagcgccgct gagaaaaagc gaagcggcac tgctctttaa
      301 caatttatca gacaatctgt gtgggcactc gaccggaatt atcgattaac tttattatta
      361 aaaattaaag aggtatatat taatgtatcg attaaataag gaggaataaa ccatgggggg
      421 ttctcatcat catcatcatc atggtatggc tagcatgact ggtggacagc aaatgggtcg
      481 ggatctgtac gacgatgacg ataaggatcc gagctcgaga tctgcagctg gtaccatatg
      541 ggaattcgaa gcttggctgt tttggcggat gagagaagat tttcagcctg atacagatta
      601 aatcagaacg cagaagcggt ctgataaaac agaatttgcc tggcggcagt agcgcggtgg
      661 tcccacctga ccccatgccg aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg
      721 ggtctcccca tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg
      781 aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca
      841 aatccgccgg gagcggattt gaacgttgcg aagcaacggc ccggagggtg gcgggcagga
      901 cgcccgccat aaactgccag gcatcaaatt aagcagaagg ccatcctgac ggatggcctt
      961 tttgcgtttc tacaaactct ttttgtttat ttttctaaat acattcaaat atgtatccgc
     1021 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     1081 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     1141 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     1201 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     1261 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg
     1321 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     1381 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     1441 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     1501 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     1561 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     1621 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     1681 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     1741 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     1801 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     1861 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     1921 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     1981 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     2041 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     2101 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     2161 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     2221 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     2281 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     2341 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     2401 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     2461 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     2521 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     2581 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     2641 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     2701 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     2761 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     2821 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     2881 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc
     2941 tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg
     3001 tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     3061 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     3121 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag cagatcaatt cgcgcgcgaa
     3181 ggcgaagcgg catgcattta cgttgacacc atcgaatggc gcaaaacctt tcgcggtatg
     3241 gcatgatagc gcccggaaga gagtcaattc agggtggtga atgtgaaacc agtaacgtta
     3301 tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag
     3361 gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc ggagctgaat
     3421 tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct gattggcgtt
     3481 gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc
     3541 gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc
     3601 tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat cattaactat
     3661 ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt tccggcgtta
     3721 tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca tgaagacggt
     3781 acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg
     3841 ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa atatctcact
     3901 cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat gtccggtttt
     3961 caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct ggttgccaac
     4021 gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg
     4081 gatatctcgg tagtgggata cgacgatacc gaagacagct catgttatat cccgccgtca
     4141 accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa
     4201 ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga
     4261 aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     4321 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     4381 tgtgagttag cgcgaattga tctg
//

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