pTrcHis C圖譜序列抗性價(jià)格Biovector質(zhì)粒庫(kù)
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pTrcHisC載體基本信息
| Invitrogen |
| pTrcHisC, pTrcHis C |
| 大腸桿菌蛋白表達(dá)載體 |
| 高拷貝 |
| Trc |
| 多克隆位點(diǎn)，限制性內(nèi)切酶 |
| 4412 bp |
| pTrcHis-F: gaggtatatattaatgtatcg |
| pTrcHis-R:GATTTAATCTGTATCAGG |
| N-His, N-EK |
| Ampicillin |
Hosts: E.coli NM522, E.coli INValphaF'. Related vectors: pSE420. |
| V360-20 |
| 瞬時(shí)表達(dá) Transient |
| 組成型 Constitutive |
| 非病毒 |
pTrcHisC載體質(zhì)粒圖譜和多克隆位點(diǎn)信息
pTrcHisC載體簡(jiǎn)介
pTrcHis載體是來(lái)由pBR322質(zhì)粒衍生而來(lái)的表達(dá)載體,設(shè)計(jì)用來(lái)在大腸桿菌中進(jìn)行重組蛋白的高效表達(dá)和純化。
載體上的Trc啟動(dòng)子和rrnB抗終止區(qū)使得高水平蛋白表達(dá)成為可能.Trc啟動(dòng)子由Trp啟動(dòng)子的-35區(qū)域和Lac啟動(dòng)子的-10區(qū)域共同組成。pTrcHis載體同時(shí)包含有一個(gè)LacIq基因，該基因編碼了Lac啟動(dòng)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(zhuǎn)錄。
進(jìn)行蛋白表達(dá)時(shí)，大腸桿菌需要生長(zhǎng)到對(duì)數(shù)生長(zhǎng)期，然后加入1mM IPTG，去除阻遏蛋白，誘導(dǎo)表達(dá)。載體上的迷你順?lè)醋咏Y(jié)構(gòu)能夠增強(qiáng)插入基因的轉(zhuǎn)錄水平，從而高水平表達(dá)目的蛋白。DNA插入片段位于載體表達(dá)片段下游，在其上游有一個(gè)N-端融合表達(dá)多肽序列，該序列包含有ATG起始密碼子，6xHis標(biāo)簽，T7噬菌體10號(hào)基因的翻譯增強(qiáng)子，X press抗原表位，腸激酶酶切識(shí)別位點(diǎn)。
pTrcHisC載體序列
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
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