Basic Vector Information
- Vector Name:
- pLenti-EF1a-C-Myc-DDK-IRES-Puro
- Antibiotic Resistance:
- Chloramphenicol
- Length:
- 8139 bp
- Type:
- Viral Expression & Packaging Vectors
- Replication origin:
- ori
- Source/Author:
- OriGene
- Copy Number:
- High copy number
- Promoter:
- EF-1α
pLenti-EF1a-C-Myc-DDK-IRES-Puro vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
pLenti-EF1a-C-Myc-DDK-IRES-Puro vector Sequence
LOCUS pLenti-EF1a-C-My 8139 bp DNA circular SYN 01-JAN-1980
DEFINITION Lentiviral PrecisionShuttle(TM) vector with a puromycin resistance
marker, for expression driven by the EF-1-alpha promoter of an ORF
fused to C-terminal c-Myc and FLAG(R) tags.
ACCESSION .
VERSION .
KEYWORDS pLenti-EF1a-C-Myc-DDK-IRES-Puro.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8139)
AUTHORS OriGene
TITLE Direct Submission
REFERENCE 2 (bases 1 to 8139)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
COMMENT Inserts are typically cloned between the SgfI (AsiSI) and MluI
sites.
FEATURES Location/Qualifiers
source 1..8139
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 237..616
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 617..819
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
LTR 834..1014
/label=5' LTR (truncated)
/note="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 1061..1186
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1683..1916
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 2101..2145
/label=gp41 peptide
/note="antigenic peptide corresponding to amino acids 655
to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
al., 2013)"
CDS 2294..2335
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
misc_feature 2443..2560
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2625..3803
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
misc_feature 3819..3928
/label=MCS
/note="MCS"
/note="multiple cloning site"
CDS 3925..3954
/label=Myc
/note="Myc (human c-Myc proto-oncogene) epitope tag"
CDS 3973..3996
/label=FLAG
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
misc_feature 4030..4610
/label=IRES2
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 4605..5201
/label=PuroR
/note="puromycin N-acetyltransferase"
protein_bind complement(5234..5267)
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
misc_feature 5323..5911
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(5914..5930)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
LTR 6440..6620
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
rep_origin complement(6682..7270)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7350..8006)
/label=CmR
/note="chloramphenicol acetyltransferase"
promoter complement(8007..8111)
/label=AmpR promoter
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