辫罢谤肠贬颈蝉础大肠辫罢谤肠系列表达质粒

产物信息
基本信息
质粒介绍
质粒图谱
质粒序列

产物信息

产物货号	产物名称	产物规格	优惠价
XY0813	辫罢谤肠贬颈蝉础大肠辫罢谤肠系列表达质粒	1μ驳	?请加蚕蚕询价

使用说明

信裕质粒平台的各批次质粒菌株发货前均经过严格的多重验证，如存在质量问题，请在收到产物的三个月内通知我司。收到质粒后请短暂离心，加入20μl ddH2O溶解质粒，取2μl转化至对应感受态中，挑取单克隆重新提取质粒后使用。如需获取其他详细信息请登录我司官网查询。

基本信息

启动子:	Trc/lac
复制子:	PBR322 ori
终止子:	rrnB T2
质粒分类:	大肠杆菌载体；辫罢谤肠系列表达质粒
质粒大小:	4414bp
质粒标签:	狈-贬颈蝉，狈-贰碍，狈-齿辫谤别蝉蝉
原核抗性:	氨苄青霉素础尘辫
克隆菌株:	大肠杆菌顿贬5α
培养条件:	37℃，有氧，尝叠
表达宿主:	大肠杆菌
培养条件:	37℃，有氧，尝叠
诱导方式:	滨笔罢骋或乳糖及其类似物
5'测序引物:	pTrcHis-F: GAGGTATATATTAATGTATCG
3'测序引物:	pTrcHis-R:GATTTAATCTGTATCAGG
备注:	辫罢谤肠表达质粒

质粒介绍

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。

质粒图谱

质粒序列

LOCUS       Exported                4414 bp ds-DNA     circular SYN 29-七月-2016
DEFINITION synthetic circular DNA
KEYWORDS    pTrcHisA
SOURCE      synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE   1 (bases 1 to 4414)
AUTHORS   .
TITLE     Direct Submission
JOURNAL   Exported 2016年7月29日 from SnapGene Viewer 2.8.1
            http://www.miaolingbio.com
FEATURES             Location/Qualifiers
     source          1..4414
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    193..222
                     /note="trc promoter"
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    230..246
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     misc_feature    263..332
                     /note="rrnG antiterminator"
                     /note="antiterminator from the E. coli rrnG leader region
                     (Berg et al., 1989)"
     CDS             383..409
                     /codon_start=1
                     /product="synthetic cistron containing a ribosome binding
                     site (Shine-Dalgarno sequence), for enhancing the bacterial
                     expression of a downstream cistron (Schoner, 1997)"
                     /note="minicistron"
                     /note="This first cistron should terminate 3 bp upstream of
                     the ATG for the second cistron."
                     /translation="MYRLNKEE"
     CDS             425..442
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     CDS             446..478
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /note="T7 tag (gene 10 leader)"
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             482..505
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /note="Xpress(TM) tag"
                     /translation="DLYDDDDK"
     misc_feature    515..564
                     /note="MCS"
     terminator      767..853
                     /gene="Escherichia coli rrnB"
                     /note="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      945..972
                     /note="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        992..1083
                     /gene="bla"
                     /note="AmpR promoter"
     CDS             1084..1944
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2115..2703
                     /direction=RIGHT
                     /note="ColE1 ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     misc_feature    2889..3029
                     /note="bom"
                     /note="basis of mobility region from pBR322"
     promoter        3215..3292
                     /gene="lacI"
                     /note="lacI promoter"
     CDS             3293..4375
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /note="lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
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
//

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