| 出品公司: | Invitrogen |
|---|---|
| 别名: | pTrcHisA, pTrcHis A |
| 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: | 4414 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 |
买家导航
pTrcHis A载体 pTrcHisA质粒基本信息
pTrcHis A质粒图谱 pTrcHisA载体图谱和pTrcHis A载体序列 pTrcHisA质粒序列多克隆位点信息
pTrcHis A质粒 pTrcHisA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。
pTrcHis A质粒序列 pTrcHisA载体序列
LOCUS pTRCHISA 4414 bp DNA circular SYN
DEFINITION pTRCHISA
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..4414
/organism="pTRCHISA"
/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(597..614)
/label="pTrcHis_rev_primer"
misc_feature complement(597..614)
/label="pBAD_rev_primer"
terminator 647..804
/label="rrnB_terminator"
terminator 770..813
/label="rrnB_T1_terminator"
terminator 945..972
/label="rrnB_T2_terminator"
promoter 1014..1042
/label="AmpR_promoter"
gene 1084..1944
/label="Ampicillin"
/gene="Ampicillin"
CDS 1084..1944
/label="ORF frame 1"
rep_origin 2099..2718
/label="pBR322_origin"
misc_feature 3115..3137
/label="pGEX_3_primer"
misc_feature 3284..4375
/label="lacI"
CDS 3416..4375
/label="ORF frame 2"
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 atggggatcc gagctcgaga tctgcagctg
541 gtaccatatg ggaattcgaa gcttggctgt tttggcggat gagagaagat tttcagcctg
601 atacagatta aatcagaacg cagaagcggt ctgataaaac agaatttgcc tggcggcagt
661 agcgcggtgg tcccacctga ccccatgccg aactcagaag tgaaacgccg tagcgccgat
721 ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa
781 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct
841 gagtaggaca aatccgccgg gagcggattt gaacgttgcg aagcaacggc ccggagggtg
901 gcgggcagga cgcccgccat aaactgccag gcatcaaatt aagcagaagg ccatcctgac
961 ggatggcctt tttgcgtttc tacaaactct ttttgtttat ttttctaaat acattcaaat
1021 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
1081 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
1141 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
1201 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
1261 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
1321 tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
1381 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
1441 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
1501 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
1561 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
1621 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
1681 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
1741 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
1801 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
1861 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
1921 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
1981 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
2041 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
2101 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
2161 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
2221 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
2281 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
2341 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
2401 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
2461 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
2521 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
2581 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
2641 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
2701 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
2761 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
2821 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
2881 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
2941 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc
3001 tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc
3061 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
3121 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag cagatcaatt
3181 cgcgcgcgaa ggcgaagcgg catgcattta cgttgacacc atcgaatggc gcaaaacctt
3241 tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga atgtgaaacc
3301 agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt
3361 ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc
3421 ggagctgaat tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct
3481 gattggcgtt gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat
3541 taaatctcgc gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg
3601 cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat
3661 cattaactat ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt
3721 tccggcgtta tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca
3781 tgaagacggt acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc
3841 gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa
3901 atatctcact cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat
3961 gtccggtttt caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct
4021 ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg
4081 cgttggtgcg gatatctcgg tagtgggata cgacgatacc gaagacagct catgttatat
4141 cccgccgtca accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg
4201 cttgctgcaa ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact
4261 ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
4321 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
4381 acgcaattaa tgtgagttag cgcgaattga tctg
//
