pTrcHis C

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pTrcHisC载体基本信息

出品公司: Invitrogen
别名: pTrcHisC, pTrcHis C
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: 4412 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.
产品目录号: V360-20
稳定性: 瞬时表达 Transient
组成型: 组成型 Constitutive
病毒/非病毒: 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
 1141 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
 1201 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
 1261 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
 1321 ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
 1381 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
 1441 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
 1501 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
 1561 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
 1621 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
 1681 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
 1741 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
 1801 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
 1861 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
 1921 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
 1981 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
 2041 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
 2101 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
 2161 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
 2221 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
 2281 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
 2341 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
 2401 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
 2461 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
 2521 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
 2581 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
 2641 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
 2701 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
 2761 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
 2821 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
 2881 agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg
 2941 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta
 3001 tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc
 3061 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
 3121 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagca gatcaattcg
 3181 cgcgcgaagg cgaagcggca tgcatttacg ttgacaccat cgaatggcgc aaaacctttc
 3241 gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag
 3301 taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg
 3361 tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg
 3421 agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga
 3481 ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta
 3541 aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg
 3601 tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca
 3661 ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc
 3721 cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg
 3781 aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc
 3841 tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat
 3901 atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt
 3961 ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg
 4021 ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg
 4081 ttggtgcgga tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc
 4141 cgccgtcaac caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct
 4201 tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg
 4261 tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
 4321 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
 4381 gcaattaatg tgagttagcg cgaattgatc tg
//

pTrcHisC载体质粒图谱pdf版和相关资料下载

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