Basic Vector Information
- Vector Name:
- pLenti-EF1a-MCS-FLAG-6×His-CMV-CopGFP-Puro
- Antibiotic Resistance:
- Ampicillin
- Length:
- 9353 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Selection Marker:
- Puro
- Promoter:
- EF-1α
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
pLenti-EF1a-MCS-FLAG-6×His-CMV-CopGFP-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-MCS-FLAG-6×His-CMV-CopGFP-Puro vector Sequence
LOCUS 62056_13840 9353 bp DNA circular SYN 01-JAN-1980
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9353)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..9353
/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"
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 2624..3802
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
CDS 3919..3942
/label=FLAG
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
CDS 3952..3969
/label=6xHis
/note="6xHis affinity tag"
promoter 4413..4612
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 4641..4659
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 4701..5366
/label=CopGFP
/note="green fluorescent protein 2 from Pontellina plumata,
also known as ppluGFP2 (Shagin et al., 2004)"
CDS 5444..5500
/label=P2A
/note="2A peptide from porcine teschovirus-1 polyprotein"
CDS 5501..6097
/label=PuroR
/note="puromycin N-acetyltransferase"
protein_bind complement(6153..6186)
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
misc_feature 6242..6830
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(6833..6849)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
LTR 7359..7539
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
rep_origin complement(7601..8189)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(8363..9220)
/label=AmpR
/note="beta-lactamase"
promoter complement(9221..9325)
/label=AmpR promoter
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