出品公司: | Invitrogen |
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別名: | pTrcHisB, pTrcHis B |
質粒類型: | 大腸桿菌蛋白表達載體 |
表達水平: | 高拷貝 |
啟動子: | Trc |
克隆方法: | 多克隆位點,限制性內切酶 |
載體大小: | 4404 bp |
5' 測序引物及序列: | pTrcHis-F: gaggtatatattaatgtatcg |
3' 測序引物及序列: | pTrcHis-R: GATTTAATCTGTATCAGG |
載體標簽: | N-His, N-EK |
載體抗性: | Ampicillin |
備注: |
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產品目錄號: | V360-20 |
穩定性: | 瞬時表達 Transient |
組成型: | 組成型 Constitutive |
病毒/非病毒: | 非病毒 |
買家導航
pTrcHis B載體 pTrcHisB質粒基本信息
pTrcHis B質粒圖譜 pTrcHisB載體圖譜和pTrcHis B載體序列 pTrcHisB質粒序列多克隆位點信息
pTrcHis B質粒 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。
pTrcHis B質粒序列 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 //gt12365.cn/ COMMENT ORIGDB|GenBank COMMENT VNTAUTHORNAME|gt12365.cn| 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 //