LOCUS       Exported                7400 bp ds-DNA    circular SYN 09-3-2013
DEFINITION  Vector for expressing the tTA transactivator in the Tet-Off(R) 
            system.
ACCESSION   .
VERSION     .
KEYWORDS    pTet-Off
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7400)
  AUTHORS   Resnitzky D, Gossen M, Bujard H, Reed SI.
  TITLE     Acceleration of the G1/S phase transition by expression of cyclins 
            D1 and E with an inducible system.
  JOURNAL   Mol. Cell. Biol. 1994;14:1669-79.
  PUBMED    8114703
REFERENCE   2  (bases 1 to 7400)
  AUTHORS   Clontech
  TITLE     Direct Submission
COMMENT     Also known as pUHD15-1neo.
FEATURES             Location/Qualifiers
     source          1..7400
                     /organism="synthetic DNA construct"
                     /lab_host="Mammalian Cells"
                     /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             774..1781
                     /codon_start=1
                     /product="tetracycline-controlled transactivator, 
                     comprising a fusion of the tetracycline repressor TetR with
                     the C-terminal activation domain of herpes simplex virus 
                     VP16"
                     /note="tTA"
                     /note="In the Tet-Off(R) system, tTA binds to the 
                     tetracycline response element and stimulates transcription 
                     only in the absence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGSAYSRARTKNNYGSTI
                     EGLLDLPDDDAPEEAGLAAPRLSFLPAGHTRRLSTAPPTDVSLGDELHLDGEDVAMAHA
                     DALDDFDLDMLGDGDSPGPGFTPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     polyA_signal    1792..1926
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2646..3234)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3405..4265)
                     /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(4266..4370)
                     /gene="bla"
                     /note="AmpR promoter"
     polyA_signal    4483..4617
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     intron          5192..5257
                     /note="small t intron"
                     /note="simian virus 40 (SV40) small t antigen intron"
     CDS             complement(5677..6471)
                     /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)"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(6832..7161)
                     /note="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     rep_origin      6846..6981
                     /note="SV40 ori"
                     /note="SV40 origin of replication"