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pTrcHisB載體基本信息
| Invitrogen |
| pTrcHisB, pTrcHis B |
| 大腸桿菌蛋白表達載體 |
| 高拷貝 |
| Trc |
| 多克隆位點，限制性內切酶 |
| 4404 bp |
| pTrcHis-F: gaggtatatattaatgtatcg |
| pTrcHis-R: GATTTAATCTGTATCAGG |
| N-His, N-EK |
| Ampicillin |
| V360-20 |
| 瞬時表達 Transient |
| 組成型 Constitutive |
| 非病毒 |
pTrcHisB載體質粒圖譜和多克隆位點信息
pTrcHisB載體簡介
pTrcHis載體是來由pBR322質粒衍生而來的表達載體,設計用來在大腸桿菌中進行重組蛋白的高效表達和純化。
載體上的Trc啟動子和rrnB抗終止區使得高水平蛋白表達成為可能.Trc啟動子由Trp啟動子的-35區域和Lac啟動子的-10區域共同組成。pTrcHis載體同時包含有一個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錄。
進行蛋白表達時，大腸桿菌需要生長到對數生長期，然后加入1mM IPTG，去除阻遏蛋白，誘導表達。載體上的迷你順反子結構能夠增強插入基因的轉錄水平，從而高水平表達目的蛋白。DNA插入片段位于載體表達片段下游，在其上游有一個N-端融合表達多肽序列，該序列包含有ATG起始密碼子，6xHis標簽，T7噬菌體10號基因的翻譯增強子，X press抗原表位，腸激酶酶切識別位點。
pTrcHisB載體序列
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
//
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