Plasmid classification: mammalian cells, tetracycline system vector.
pRevTRE is a retroviral Tet response vector that expresses a gene of interest from the Tetresponse element (TRE). This vector is derived from pLNCX, a retroviral vector created using elements of Moloney murine leukemia virus (MoMuLV) and Moloney murine sarcoma virus (MoMuSV) as described (1). The TRE contains seven direct repeats of the tetO operator sequence, upstream of a minimal CMV promoter, which can be bound by the tTA and rtTA transactivators. The 5' viral LTR controls expression of the transcript that contains 唯+ (the extended viral packaging signal) and the hygromycin resistance (Hygr) gene for antibiotic selection in mammalian cells. The TRE is derived from vectors described previously (2, 3). pRevTRE also includes the E. coli Ampr gene for antibiotic selection in bacteria.
pRevTRE閳?Sequence
LOCUS Exported 6487 bp ds-DNA circular SYN 05-SEP-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled 4
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6487)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Monday, September 5, 2016 from SnapGene Viewer 3.1.4
FEATURES Location/Qualifiers
source 1..6487
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 414..589
/note="5' LTR (truncated)"
/note="truncated long terminal repeat from Moloney murine
sarcoma virus"
misc_feature 652..851
/note="MMLV Psi"
/note="packaging signal of Moloney murine leukemia virus
(MMLV)"
CDS 1052..1468
/note="gag (truncated)"
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
CDS 1524..2543
/codon_start=1
/gene="aph(4)-Ia"
/product="aminoglycoside phosphotransferase from E. coli"
/note="HygR"
/note="confers resistance to hygromycin"
/translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
ELPAVLQPVAEAAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
protein_bind 2847..3117
/bound_moiety="tetracycline repressor TetR"
/note="tetracycline response element"
/note="contains seven copies of the tetracycline operator
tetO"
promoter 3150..3187
/note="minimal CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
misc_feature 3281..3337
/note="MCS"
/note="pUC18/19 multiple cloning site"
LTR 3389..3982
/note="long terminal repeat from Moloney murine leukemia
virus"
misc_feature 4193..4333
/note="bom"
/note="basis of mobility region from pBR322"
rep_origin complement(4519..5107)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5278..6138)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(6139..6243)
/gene="bla"
/note="AmpR promoter"
ORIGIN
1 tttgaaagac cccacccgta ggtggcaagc tagcttaagt aacgccactt tgcaaggcat
61 ggaaaaatac ataactgaga atagaaaagt tcagatcaag gtcaggaaca aagaaacagc
121 tgaataccaa acaggatatc tgtggtaagc ggttcctgcc ccggctcagg gccaagaaca
181 gatgagacag ctgagtgatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg
241 ctcggggcca agaacagatg gtccccagat gcggtccagc cctcagcagt ttctagtgaa
301 tcatcagatg tttccagggt gccccaagga cctgaaaatg accctgtacc ttatttgaac
361 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc cgctctccga gctcaataaa
421 agagcccaca acccctcact cggcgcgcca gtcttccgat agactgcgtc gcccgggtac
481 ccgtattccc aataaagcct cttgctgttt gcatccgaat cgtggtctcg ctgttccttg
541 ggagggtctc ctctgagtga ttgactaccc acgacggggg tctttcattt gggggctcgt
601 ccgggatttg gagacccctg cccagggacc accgacccac caccgggagg taagctggcc
661 agcaacttat ctgtgtctgt ccgattgtct agtgtctatg tttgatgtta tgcgcctgcg
721 tctgtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa ctgacgagtt
781 ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc gtttttgtgg
841 cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg tggttctggt
901 aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt cggtttggaa
961 ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct
1021 gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc ccttaagttt
1081 gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa
1141 gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg tcggatggcc
1201 gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg tcttttcacc
1261 tggcccgcat
文献和实验
技术资料