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pCMV-Tet3G

pCMV-Tet3G
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  • 产品名称:pCMV-Tet3G
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  • 产品展商:XYbscience
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简单介绍
pCMV-Tet3G菌株发货前均经过严格的多重验证,如存在质量问题,请在收到产品的三个月内通知我司。pCMV-Tet3G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��重新提取质粒后使用。
产品描述

pCMV-Tet3G

  • 产品信息
  • 基本信息
  • 质粒简介
  • 质粒图谱
  • 质粒序列

产品信息

产品货号 产品名称 产品规格 优惠价
XY0725 pCMV-Tet3G

20μl

¥请询价

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,请在收到产品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: CMV
复制子: pUC ori,SV40 ori
终止子: SV40 poly(A) signal
质粒分类: 哺乳细胞,四环素调控系统载体
质粒大小: 7139bp
原核抗性: Amp
克隆菌株: DH5α
培养条件: 37℃,有氧 LB
表达宿主: 哺乳细胞
诱导方式: 四环素诱导
5'测序引物: CMV-F:CGCAAATGGGCGGTAGGCGTG
3'测序引物: 根据序列设计引物

质粒简介

     The pCMV-Tet3G Vector expresses Tet-On® 3G, a tetracycline-controlled transactivator that exhibits high activity in the presence of the inducer doxycycline (Dox) and exceptionally low activity in its absence. Tet-On 3G results from the fusion of amino acids 1–207 of a mutant Tet repressor (TetR) to 39 amino acids that form three minimal "F"-type transcriptional activation domains from the herpes simplex virus VP16 protein. Tet-On 3G was derived from Tet-On Advanced (1–4); as a result, it’s fully synthetic, lacks cryptic splice sites, and is codon-optimized for stable expression in mammalian cells. Compared to both of its predecessors, this 3rd generation Tet-On transactivator demonstrates increased sensitivity to Dox (1). Constitutive expression of Tet-On 3G is driven by the human cytomegalovirus immediately early promoter (PCMV IE). Note: An EF1α version of this vector is available for cell lines in which the CMV promoter is silenced.
     pCMV-Tet3G is used to develop stable Tet-On 3G cell lines, which are hosts for Tet-inducible gene expression systems. To create a Tet-inducible expression system, a vector containing a gene of interest under the control of the Tet-inducible TRE3G promoter (PTRE3G) is transfected into a Tet-On 3G cell line. The addition of Dox to the system causes Tet-On 3G to undergo a conformational change that allows it to bind to PTRE3G, activating transcription of the gene of interest in a highly dose-dependent manner

质粒图谱

质粒序列

LOCUS       Exported                7139 bp ds-DNA     circular SYN 05-SEP-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Untitled 6
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7139)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, September 5, 2016 from SnapGene Viewer 3.1.4
            http://www.miaolingbio.com
FEATURES             Location/Qualifiers
     source          1..7139
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        89..468
                     /note="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        469..672
                     /note="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             775..1521
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /note="Tet-On(R) 3G"
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     polyA_signal    1531..1665
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2385..2973)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3144..4004)
                     /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"
     promoter        complement(4005..4109)
                     /gene="bla"
                     /note="AmpR promoter"
     polyA_signal    4222..4356
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     CDS             complement(4781..4801)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T 
                     antigen"
                     /note="SV40 NLS"
                     /translation="PKKKRKV"
     intron          4931..4996
                     /note="small t intron"
                     /note="simian virus 40 (SV40) small t antigen intron"
     CDS             complement(5416..6210)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /note="NeoR/KanR"
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(6571..6900)
                     /note="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     rep_origin      6585..6720
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
ORIGIN
        1 ctcgaggagc ttggcccatt gcatacgttg tatccatatc ataatatgta catttatatt
       61 ggctcatgtc caacattacc gccatgttga cattgattat tgactagtta ttaatagtaa
      121 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
      181 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
      241 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
      301 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
      361 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
      421 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
      481 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
      541 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
      601 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
      661 ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt
      721 gacctccata gaagacaccg ggaccgatcc agcctccgcg gccccgaatt caccatgtct
      781 agactggaca agagcaaagt cataaactct gctctggaat tactcaatgg agtcggtatc
      841 gaaggcctga cgacaaggaa actcgctcaa aagctgggag ttgagcagcc taccctgtac
      901 tggcacgtga agaacaagcg ggccctgctc gatgccctgc caatcgagat gctggacagg
      961 catcataccc actcctgccc cctggaaggc gagtcatggc aagactttct gcggaacaac
     1021 gccaagtcat accgctgtgc tctcctctca catcgcgacg gggctaaagt gcatctcggc
     1081 acccgcccaa cagagaaaca gtacgaaacc ctggaaaatc agctcgcgtt cctgtgtcag
     1141 caaggcttct ccctggagaa cgcactgtac gctctgtccg ccgtgggcca ctttacactg
     1201 ggctgcgtat tggaggaaca ggagcatcaa gtagcaaaag aggaaagaga gacacctacc
     1261 accgattcta tgcccccact tctgaaacaa gcaattgagc tgttcgaccg gcagggagcc
     1321 gaacctgcct tccttttcgg cctggaacta atcatatgtg gcctggagaa acagctaaag
     1381 tgcgaaagcg gcgggccgac cgacgccctt gacgattttg acttagacat gctcccagcc
     1441 gatgcccttg acgactttga ccttgatatg ctgcctgctg acgctcttga cgattttgac
     1501 cttgacatgc tccccgggta actaagtaag gatccagaca tgataagata cattgatgag
     1561 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     1621 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     1681 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
     1741 ctctacaaat gtggtatggc tgattatgat cctgcaagcc tcgtcgtcct ggccggacca
     1801 cgctatctgt gcaaggtccc cggccccgga cgcgcgctcc atgagcagag cgcccgccgc
     1861 cgaggcgaag actcgggcgg cgccctgccc gtcccaccag gtcaacaggc ggtaaccggc
     1921 ctcttcatcg ggaatgcgcg cgaccttcag catcgccggc atgtccccct ggcggacggg
     1981 aagtatccag ctcgaccaag cttggcgaga ttttcaggag ctaaggaagc taaaatggag
     2041 aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt
     2101 gaggcatttc agtcagttgc tcaatgtacc tataaccaga ccgttcagct gcattaatga
     2161 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     2221 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     2281 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     2341 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     2401 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     2461 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     2521 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     2581 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     2641 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     2701 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     2761 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     2821 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     2881 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     2941 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     3001 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     3061 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     3121 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     3181 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     3241 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     3301 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     3361 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     3421 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     3481 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     3541 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     3601 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     3661 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     3721 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca
     3781 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     3841 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     3901 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     3961 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     4021 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     4081 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     4141 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     4201 cgtcttcact cgaggtcgag ggatccagac atgataagat acattgatga gtttggacaa
     4261 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     4321 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     4381 atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa
     4441 tgtggtatgg ctgattatga tctctagtca aggcactata catcaaatat tccttattaa
     4501 cccctttaca aattaaaaag ctaaaggtac acaatttttg agcatagtta ttaatagcag
     4561 acactctatg cctgtgtgga gtaagaaaaa acagtatgtt atgattataa ctgttatgcc
     4621 tacttataaa ggttacagaa tatttttcca taattttctt gtatagcagt gcagcttttt
     4681 cctttgtggt gtaaatagca aagcaagcaa gagttctatt actaaacaca gcatgactca
     4741 aaaaacttag caattctgaa ggaaagtcct tggggtcttc tacctttctc ttcttttttg
     4801 gaggagtaga atgttgagag tcagcagtag cctcatcatc actagatggc atttcttctg
     4861 agcaaaacag gttttcctca ttaaaggcat tccaccactg ctcccattca tcagttccat
     4921 aggttggaat ctaaaataca caaacaatta gaatcagtag tttaacacat tatacactta
     4981 aaaattttat atttacctta gagctttaaa tctctgtagg tagtttgtcc aattatgtca
     5041 caccacagaa gtaaggttcc ttcacaaaga tccggaccaa agcggccatc gtgcctcccc
     5101 actcctgcag ttcgggggca tggatgcgcg gatagccgct gctggtttcc tggatgccga
     5161 cggatttgca ctgccggtag aactccgcga ggtcgtccag cctcaggcag cagctgaacc
     5221 aactcgcgag gggatcgagc ccggggtggg cgaagaactc cagcatgaga tccccgcgct
     5281 ggaggatcat ccagccggcg tcccggaaaa cgattccgaa gcccaacctt tcatagaagg
     5341 cggcggtgga atcgaaatct cgtgatggca ggttgggcgt cgcttggtcg gtcatttcga
     5401 accccagagt cccgctcaga agaactcgtc aagaaggcga tagaaggcga tgcgctgcga
     5461 atcgggagcg gcgataccgt aaagcacgag gaagcggtca gcccattcgc cgccaagctc
     5521 ttcagcaata tcacgggtag ccaacgctat gtcctgatag cggtccgcca cacccagccg
     5581 gccacagtcg atgaatccag aaaagcggcc attttccacc atgatattcg gcaagcaggc
     5641 atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg cgcgccttga gcctggcgaa
     5701 cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat cgacaagacc
     5761 ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca
     5821 ggtagccgga tcaagcgtat gcagccgccg cattgcatca gccatgatgg atactttctc
     5881 ggcaggagca aggtgagatg acaggagatc ctgccccggc acttcgccca atagcagcca
     5941 gtcccttccc gcttcagtga caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc
     6001 cagccacgat agccgcgctg cctcgtcctg cagttcattc agggcaccgg acaggtcggt
     6061 cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg aacacggcgg catcagagca
     6121 gccgattgtc tgttgtgccc agtcatagcc gaatagcctc tccacccaag cggccggaga
     6181 acctgcgtgc aatccatctt gttcaatcat gcgaaacgat cctcatcctg tctcttgatc
     6241 agatcttgat cccctgcgcc atcagatcct tggcggcaag aaagccatcc agtttacttt
     6301 gcagggcttc ccaaccttac cagagggcgc cccagctggc aattccggtt cgcttgctgt
     6361 ccataaaacc gcccagtcta gctatcgcca tgtaagccca ctgcaagcta cctgctttct
     6421 ctttgcgctt gcgttttccc ttgtccagat agcccagtag ctgacattca tccggggtca
     6481 gcaccgtttc tgcggactgg ctttctacgt gttccgcttc ctttagcagc ccttgcgccc
     6541 tgagtgcttg cggcagcgtg aagctagctt tttgcaaaag cctaggcctc caaaaaagcc
     6601 tcctcactac ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa
     6661 aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat
     6721 gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcatgc tttgcatact
     6781 tctgcctgct ggggagcctg gggactttcc acacctggtt gctgactaat tgagatgcat
     6841 gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac
     6901 attccacagc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     6961 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     7021 cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag
     7081 cggagtgtat actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcacct
//
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