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 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 cgattttga <
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