pTrcHis A图谱序列抗性价格Biovector质粒库

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pTrcHisA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

pTrcHisA, pTrcHis A

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

4414 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

pTrcHisA载体质粒图谱和多克隆位点信息

pTrcHisA 载体图谱

pTrcHisA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。

pTrcHisA载体序列

LOCUS       pTRCHISA   4414 bp    DNA circular    SYN
DEFINITION pTRCHISA
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..4414
                     /organism="pTRCHISA"
                     /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(597..614)
                     /label="pTrcHis_rev_primer"
     misc_feature    complement(597..614)
                     /label="pBAD_rev_primer"
     terminator      647..804
                     /label="rrnB_terminator"
     terminator      770..813
                     /label="rrnB_T1_terminator"
     terminator      945..972
                     /label="rrnB_T2_terminator"
     promoter        1014..1042
                     /label="AmpR_promoter"
     gene            1084..1944
                     /label="Ampicillin"
                     /gene="Ampicillin"
     CDS             1084..1944
                     /label="ORF frame 1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
     rep_origin      2099..2718
                     /label="pBR322_origin"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
     misc_feature    3115..3137
                     /label="pGEX_3_primer"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
     misc_feature    3284..4375
                     /label="lacI"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
     CDS             3416..4375
                     /label="ORF frame 2"
                     /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 atggggatcc gagctcgaga tctgcagctg
541 gtaccatatg ggaattcgaa gcttggctgt tttggcggat gagagaagat tttcagcctg
601 atacagatta aatcagaacg cagaagcggt ctgataaaac agaatttgcc tggcggcagt
661 agcgcggtgg tcccacctga ccccatgccg aactcagaag tgaaacgccg tagcgccgat
721 ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa
781 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct
841 gagtaggaca aatccgccgg gagcggattt gaacgttgcg aagcaacggc ccggagggtg
901 gcgggcagga cgcccgccat aaactgccag gcatcaaatt aagcagaagg ccatcctgac
961 ggatggcctt tttgcgtttc tacaaactct ttttgtttat ttttctaaat acattcaaat
1021 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
1081 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
&nb