pCMV- Tet3G

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  • Model: PVT11010
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pCMV-Tet3G

PVT11010  2ug

 

pCMV-Tet3G Description

pCMV-Tet3G is a Mammal plasmid.

 

pCMV-Tet3G Informaiton

Promoter: CMV

Replicon: pUC ori, SV40 ori

Terminator: SV40 poly (A) signal

Plasmid classification: mammalian cells, tetracycline regulatory system vectors

Plasmid size: 7139bp

Prokaryotic resistance: Amp

Clonal strain: DH5 alpha

Culture conditions: 37 C, aerobic LB

Expression host: lactation cells

Induction: tetracycline induction

5'sequencing primers: CMV-F:CGCAAATGGGCGGTAGGCGTG

Primers for 3'sequencing: primers designed based on sequences

 

pCMV-Tet3G Introduction

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.

 

pCMV-Tet3G Multiple cloning site

pCMV-Tet3G-vector

 

 

 

pCMV-Tet3G Sequence

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

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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Product is for research use only!

 

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