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pTrcHisB载体基本信息
| Invitrogen |
| pTrcHisB, pTrcHis B |
| 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 |
| 4404 bp |
| pTrcHis-F: gaggtatatattaatgtatcg |
| pTrcHis-R: GATTTAATCTGTATCAGG |
| N-His, N-EK |
| Ampicillin |
| V360-20 |
| 瞬时表达 Transient |
| 组成型 Constitutive |
| 非病毒 |
pTrcHisB载体质粒图谱和多克隆位点信息
pTrcHisB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。
pTrcHisB载体序列
LOCUS pTrcHis\B 4404 bp DNA circular SYN
DEFINITION pTRCHISB
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..4404
/organism="pTRCHISB"
/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(587..604)
/label="pTrcHis_rev_primer"
misc_feature complement(587..604)
/label="pBAD_rev_primer"
terminator 637..794
/label="rrnB_terminator"
terminator 760..803
/label="rrnB_T1_terminator"
terminator 935..962
/label="rrnB_T2_terminator"
promoter 1004..1032
/label="AmpR_promoter"
gene 1074..1934
/label="Ampicillin"
/gene="Ampicillin"
CDS 1074..1934
/label="ORF frame 3"
rep_origin 2089..2708
/label="pBR322_origin"
misc_feature 3105..3127
/label="pGEX_3_primer"
misc_feature 3274..4365
/label="lacI"
CDS 3406..4365
/label="ORF frame 1"
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 ataaggatcc gagctcgaga tctgcagctg gtaccatatg
541 ggaattcgaa gcttggctgt tttggcggat gagagaagat tttcagcctg atacagatta
601 aatcagaacg cagaagcggt ctgataaaac agaatttgcc tggcggcagt agcgcggtgg
661 tcccacctga ccccatgccg aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg
721 ggtctcccca tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg
781 aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca
841 aatccgccgg gagcggattt gaacgttgcg aagcaacggc ccggagggtg gcgggcagga
901 cgcccgccat aaactgccag gcatcaaatt aagcagaagg ccatcctgac ggatggcctt
961 tttgcgtttc tacaaactct ttttgtttat ttttctaaat acattcaaat atgtatccgc
1021 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
1081 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
1141 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
1201 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
1261 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg
1321 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
1381 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
1441 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
1501 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
1561 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
1621 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
1681 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
1741 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
1801 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
1861 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
1921 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
1981 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
2041 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
2101 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
2161 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
2221 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
2281 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
2341 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
2401 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
2461 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
2521 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
2581 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
2641 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
2701 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
2761 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
2821 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
2881 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc
2941 tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg
3001 tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
3061 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
3121 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag cagatcaatt cgcgcgcgaa
3181 ggcgaagcgg catgcattta cgttgacacc atcgaatggc gcaaaacctt tcgcggtatg
3241 gcatgatagc gcccggaaga gagtcaattc agggtggtga atgtgaaacc agtaacgtta
3301 tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag
3361 gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc ggagctgaat
3421 tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct gattggcgtt
3481 gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc
3541 gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc
3601 tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat cattaactat
3661 ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt tccggcgtta
3721 tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca tgaagacggt
3781 acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg
3841 ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa atatctcact
3901 cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat gtccggtttt
3961 caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct ggttgccaac
4021 gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg
4081 gatatctcgg tagtgggata cgacgatacc gaagacagct catgttatat cccgccgtca
4141 accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa
4201 ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga
4261 aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
4321 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
4381 tgtgagttag cgcgaattga tctg
//
DEFINITION pTRCHISB
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..4404
/organism="pTRCHISB"
/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(587..604)
/label="pTrcHis_rev_primer"
misc_feature complement(587..604)
/label="pBAD_rev_primer"
terminator 637..794
/label="rrnB_terminator"
terminator 760..803
/label="rrnB_T1_terminator"
terminator 935..962
/label="rrnB_T2_terminator"
promoter 1004..1032
/label="AmpR_promoter"
gene 1074..1934
/label="Ampicillin"
/gene="Ampicillin"
CDS 1074..1934
/label="ORF frame 3"
rep_origin 2089..2708
/label="pBR322_origin"
misc_feature 3105..3127
/label="pGEX_3_primer"
misc_feature 3274..4365
/label="lacI"
CDS 3406..4365
/label="ORF frame 1"
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 ataaggatcc gagctcgaga tctgcagctg gtaccatatg
541 ggaattcgaa gcttggctgt tttggcggat gagagaagat tttcagcctg atacagatta
601 aatcagaacg cagaagcggt ctgataaaac agaatttgcc tggcggcagt agcgcggtgg
661 tcccacctga ccccatgccg aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg
721 ggtctcccca tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg
781 aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca
841 aatccgccgg gagcggattt gaacgttgcg aagcaacggc ccggagggtg gcgggcagga
901 cgcccgccat aaactgccag gcatcaaatt aagcagaagg ccatcctgac ggatggcctt
961 tttgcgtttc tacaaactct ttttgtttat ttttctaaat acattcaaat atgtatccgc
1021 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
1081 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
1141 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
1201 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
1261 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg
1321 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
1381 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
1441 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
1501 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
1561 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
1621 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
1681 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
1741 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
1801 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
1861 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
1921 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
1981 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
2041 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
2101 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
2161 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
2221 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
2281 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
2341 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
2401 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
2461 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
2521 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
2581 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
2641 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
2701 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
2761 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
2821 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
2881 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc
2941 tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg
3001 tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
3061 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
3121 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag cagatcaatt cgcgcgcgaa
3181 ggcgaagcgg catgcattta cgttgacacc atcgaatggc gcaaaacctt tcgcggtatg
3241 gcatgatagc gcccggaaga gagtcaattc agggtggtga atgtgaaacc agtaacgtta
3301 tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag
3361 gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc ggagctgaat
3421 tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct gattggcgtt
3481 gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc
3541 gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc
3601 tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat cattaactat
3661 ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt tccggcgtta
3721 tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca tgaagacggt
3781 acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg
3841 ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa atatctcact
3901 cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat gtccggtttt
3961 caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct ggttgccaac
4021 gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg
4081 gatatctcgg tagtgggata cgacgatacc gaagacagct catgttatat cccgccgtca
4141 accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa
4201 ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga
4261 aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
4321 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
4381 tgtgagttag cgcgaattga tctg
//
pTrcHisB载体质粒图谱pdf版和相关资料下载
pTrcHisB载体应用举例
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