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pTrcHis C图谱序列抗性价格Biovector质粒库

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pTrcHisC载体基本信息

出品公司:别名:质粒类型:表达水平:启动子:克隆方法:载体大小:5' 测序引物及序列:3' 测序引物及序列:载体标签:载体抗性:备注:产品目录号:稳定性:组成型:病毒/非病毒:
Invitrogen
pTrcHisC, pTrcHis C
大肠杆菌蛋白表达载体
高拷贝
Trc
多克隆位点,限制性内切酶
4412 bp
pTrcHis-F: gaggtatatattaatgtatcg
pTrcHis-R:GATTTAATCTGTATCAGG
N-His, N-EK
Ampicillin
Hosts: E.coli NM522, E.coli INValphaF'. Related vectors: pSE420.
V360-20
瞬时表达 Transient
组成型 Constitutive
非病毒

pTrcHisC载体质粒图谱和多克隆位点信息

pTrcHisC载体图谱

pTrcHisC载体多克隆位点

pTrcHisC载体简介

pTrcHis载体是来由pBR322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Trc启动子和rrnB抗终止区使得高水平蛋白表达成为可能.Trc启动子由Trp启动子的-35区域和Lac启动子的-10区域共同组成。pTrcHis载体同时包含有一个LacIq基因,该基因编码了Lac启动子的抑制蛋白。LacIq基因的存在使得宿主菌是LacIq+还是LacIq-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。

pTrcHisC载体序列

LOCUS       pTrcHisC   4412 bp     DNA circular   SYN
DEFINITION  pTRCHISC
ACCESSION   
KEYWORDS    
SOURCE      
  ORGANISM  other sequences; artificial sequences; vectors.
COMMENT     This file is created by Vector NTI
            http://www.biofeng.com/
COMMENT     ORIGDB|GenBank
COMMENT     VNTAUTHORNAME|biofeng.com|
FEATURES             Location/Qualifiers
     source          1..4412
                     /organism="pTRCHISC"
                     /mol_type="other DNA"
     misc_feature    235..257
                     /label="M13_pUC_rev_primer"
     misc_feature    264..333
                     /label="rrnB_ATS"
     misc_feature    370..390
                     /label="pTrcHis_fwd_primer"
     misc_feature    383..409
                     /label="minicis"
     misc_feature    445..463
                     /label="Xpress_fwd_primer"
     misc_feature    446..478
                     /label="T7_leader"
     misc_feature    482..505
                     /label="Xpress_EK"
     misc_feature    complement(595..612)
                     /label="pBAD_rev_primer"
     misc_feature    complement(595..612)
                     /label="pTrcHis_rev_primer"
     terminator      645..802
                     /label="rrnB_terminator"
     terminator      768..811
                     /label="rrnB_T1_terminator"
     terminator      943..970
                     /label="rrnB_T2_terminator"
     promoter        1012..1040
                     /label="AmpR_promoter"
     gene            1082..1942
                     /label="Ampicillin"
                     /gene="Ampicillin"
     CDS             1082..1942
                     /label="ORF frame 2"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
     rep_origin      2097..2716
                     /label="pBR322_origin"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
     misc_feature    3113..3135
                     /label="pGEX_3_primer"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
     misc_feature    3282..4373
                     /label="lacI"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
     CDS             3414..4373
                     /label="ORF frame 3"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
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 ataaggatcg atggatccga cctcgagatc tgcagatggt
  541 accatatggg aattcgaagc ttggctgttt tggcggatga gagaagattt tcagcctgat
  601 acagattaaa tcagaacgca gaagcggtct gataaaacag aatttgcctg gcggcagtag
  661 cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg
  721 tagtgtgggg tctccccatg cgagagtagg gaactgccag gcatcaaata aaacgaaagg
  781 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga
  841 gtaggacaaa tccgccggga gcggatttga acgttgcgaa gcaacggccc ggagggtggc
  901 gggcaggacg cccgccataa actgccaggc atcaaattaa gcagaaggcc atcctgacgg
  961 atggcctttt tgcgtttcta caaactcttt ttgtttattt ttctaaatac attcaaatat
 1021 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
 1081 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
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