pTrcHis A

pTrcHis A

载体基本信息

出品公司:	Ybscience
别名:	pTrcHisA, pTrcHis A
质粒类型:	大肠杆菌蛋白表达载体
表达水平:	高拷贝
启动子:	Trc
克隆方法:	多克隆位点，限制性内切酶
载体大小:	4414 bp
5' 测序引物及序列:	pTrcHis-F: gaggtatatattaatgtatcg
3' 测序引物及序列:	pTrcHis-R:GATTTAATCTGTATCAGG
载体标签:	N-His, N-EK
载体抗性:	Ampicillin
备注:	Hosts: E.coli NM522, E.coli INValphaF'. Related vectors: pSE420.
稳定性:	瞬时表达 Transient
组成型:	组成型 Constitutive
病毒/非病毒:	非病毒

pTrcHisA载体质粒图谱和多克隆位点信息

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"
     rep_origin      2099..2718
                     /label="pBR322_origin"
     misc_feature    3115..3137
                     /label="pGEX_3_primer"
     misc_feature    3284..4375
                     /label="lacI"
     CDS             3416..4375
                     /label="ORF frame 2"
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
 1141 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
 1201 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
 1261 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
 1321 tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
 1381 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
 1441 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
 1501 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
 1561 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
 1621 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
 1681 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
 1741 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
 1801 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
 1861 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
 1921 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
 1981 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
 2041 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
 2101 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
 2161 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
 2221 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
 2281 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
 2341 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
 2401 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
 2461 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
 2521 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
 2581 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
 2641 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
 2701 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
 2761 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
 2821 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
 2881 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
 2941 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc
 3001 tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc
 3061 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
 3121 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag cagatcaatt
 3181 cgcgcgcgaa ggcgaagcgg catgcattta cgttgacacc atcgaatggc gcaaaacctt
 3241 tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga atgtgaaacc
 3301 agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt
 3361 ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc
 3421 ggagctgaat tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct
 3481 gattggcgtt gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat
 3541 taaatctcgc gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg
 3601 cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat
 3661 cattaactat ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt
 3721 tccggcgtta tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca
 3781 tgaagacggt acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc
 3841 gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa
 3901 atatctcact cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat
 3961 gtccggtttt caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct
 4021 ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg
 4081 cgttggtgcg gatatctcgg tagtgggata cgacgatacc gaagacagct catgttatat
 4141 cccgccgtca accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg
 4201 cttgctgcaa ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact
 4261 ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
 4321 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
 4381 acgcaattaa tgtgagttag cgcgaattga tctg
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