Cloning host is Stbl3 or TOP10
| 载体基本信息 | |||
| 载体名称 | LeGO-iC2 | 载体抗性 | Ampicillin |
|---|---|---|---|
| 载体长度 | 7820 bp | 载体类型 | Lentiviral vectors |
| 拷贝数 | High copy number | 克隆方法 | Enzyme digestion and ligation |
| 5引物 | GAGCTCACAACCCCTCACTC | ||
LeGO-iC2 载体载体图谱
LeGO-iC2 载体载体序列
LOCUS Exported 7820 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 7820)
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..7820
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 238..617
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 619..817
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
LTR 835..1015
/note="5 LTR (truncated)"
/note="truncated 5 long terminal repeat (LTR) from HIV-1"
misc_feature 1062..1187
/note="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1686..1919
/note="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature 2446..2563
/note="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
primer_bind 2622..2638
/note="KS primer"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 2680..2713
/bound_moiety="Cre recombinase"
/note="loxP"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
promoter 2784..3190
/note="SFFV promoter"
/note="spleen focus-forming virus long terminal repeat
(LTR) promoter"
misc_feature 3276..3862
/note="IRES2"
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 3863..4573
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/note="mCherry"
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
protein_bind 4620..4653
/bound_moiety="Cre recombinase"
/note="loxP"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
misc_feature 4709..5297
/note="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
CDS complement(5180..5191)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/note="Factor Xa site"
/translation="IEGR"
primer_bind complement(5300..5316)
/note="KS primer"
/note="common sequencing primer, one of multiple similar
variants"
LTR 5826..6006
/note="5 LTR (truncated)"
/note="truncated 5 long terminal repeat (LTR) from HIV-1"
rep_origin complement(6068..6656)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6827..7687)
/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"
promoter complement(7688..7792)
/gene="bla"
/note="AmpR promoter"
