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pTrcHis A圖譜序列抗性價(jià)格Biovector質(zhì)粒庫

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  • 貨　　號：39599964164
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pTrcHisA載體基本信息

出品公司:別名:質(zhì)粒類型:表達(dá)水平:啟動子:克隆方法:載體大小:5' 測序引物及序列:3' 測序引物及序列:載體標(biāo)簽:載體抗性:備注:產(chǎn)品目錄號:穩(wěn)定性:組成型:病毒/非病毒:
Invitrogen
pTrcHisA, pTrcHis A
大腸桿菌蛋白表達(dá)載體
高拷貝
Trc
多克隆位點(diǎn)，限制性內(nèi)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
瞬時(shí)表達(dá) Transient
組成型 Constitutive
非病毒

pTrcHisA載體質(zhì)粒圖譜和多克隆位點(diǎn)信息

pTrcHisA 載體圖譜

pTrcHisA多克隆位點(diǎn)

pTrcHisA載體簡介

pTrcHis載體是來由pBR322質(zhì)粒衍生而來的表達(dá)載體,設(shè)計(jì)用來在大腸桿菌中進(jìn)行重組蛋白的高效表達(dá)和純化。

載體上的Trc啟動子和rrnB抗終止區(qū)使得高水平蛋白表達(dá)成為可能.Trc啟動子由Trp啟動子的-35區(qū)域和Lac啟動子的-10區(qū)域共同組成。pTrcHis載體同時(shí)包含有一個(gè)LacIq基因，該基因編碼了Lac啟動子的抑制蛋白。LacIq基因的存在使得宿主菌是LacIq+還是LacIq-都可以高效抑制插入基因的轉(zhuǎn)錄。

進(jìn)行蛋白表達(dá)時(shí)，大腸桿菌需要生長到對數(shù)生長期，然后加入1mM IPTG，去除阻遏蛋白，誘導(dǎo)表達(dá)。載體上的迷你順反子結(jié)構(gòu)能夠增強(qiáng)插入基因的轉(zhuǎn)錄水平，從而高水平表達(dá)目的蛋白。DNA插入片段位于載體表達(dá)片段下游，在其上游有一個(gè)N-端融合表達(dá)多肽序列，該序列包含有ATG起始密碼子，6xHis標(biāo)簽，T7噬菌體10號基因的翻譯增強(qiáng)子，X press抗原表位，腸激酶酶切識別位點(diǎn)。

pTrcHisA載體序列

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

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