CMV promoter is upstream of the bicistronic coding cassette with SV40-Large T and HSV1-TK. During reverse transcription U3 region of the 3LTR, with LoxP, is duplicated; LoxP sites end up flanking the genome of the integrated provirus. Upon expression of Cre, provirus is excised.
| 载体基本信息 | |||
| 载体名称 | pLOX-Ttag-iresTK | 载体抗性 | Ampicillin |
|---|---|---|---|
| 载体长度 | 12290 bp | 载体类型 | Lentiviral vectors |
| 拷贝数 | High copy number | 克隆方法 | Enzyme digestion and ligation |
| 5引物 | CMV Fwd | 3引物 | SP6 rev |
pLOX-Ttag-iresTK 载体载体图谱
pLOX-Ttag-iresTK 载体载体序列
LOCUS Exported 12290 bp ds-DNA circular SYN 13-JAN-2022
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12290)
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..12290
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 4..637
/note="3 LTR"
/note="3 long terminal repeat (LTR) from HIV-1"
misc_feature 684..809
/note="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1302..1535
/note="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
enhancer 2043..2346
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 2347..2550
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
intron 3257..3322
/note="small t intron"
/note="simian virus 40 (SV40) small t antigen intron"
CDS 3452..3472
/codon_start=1
/product="nuclear localization signal of SV40 large T
antigen"
/note="SV40 NLS"
/translation="PKKKRKV"
primer_bind complement(5256..5272)
/note="KS primer"
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 5321..5782
/note="IRES"
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
protein_bind 6854..6887
/bound_moiety="Cre recombinase"
/note="loxP"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
LTR 6907..7087
/note="5 LTR (truncated)"
/note="truncated 5 long terminal repeat (LTR) from HIV-1"
promoter complement(7112..7130)
/note="SP6 promoter"
/note="promoter for bacteriophage SP6 RNA polymerase"
promoter 7918..8022
/gene="bla"
/note="AmpR promoter"
CDS 8023..8883
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 9054..9642
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 9888..10217
/note="SV40 promoter"
/note="SV40 enhancer and early promoter"
rep_origin 10068..10203
/note="SV40 ori"
/note="SV40 origin of replication"
intron 11351..11416
/note="small t intron"
/note="simian virus 40 (SV40) small t antigen intron"
CDS 11546..11566
/codon_start=1
/product="nuclear localization signal of SV40 large T
antigen"
/note="SV40 NLS"
/translation="PKKKRKV"
polyA_signal 11991..12125
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
