Basic Vector Information
- Vector Name:
- pLVX-EF1alpha-SARS-CoV-2-nsp12-2xStrep-IRES-Puro
- Antibiotic Resistance:
- Ampicillin
- Length:
- 11712 bp
- Type:
- Mammalian Expression, Lentiviral
- Replication origin:
- ori
- Selection Marker:
- Puromycin
- Promoter:
- EF-1α
pLVX-EF1alpha-SARS-CoV-2-nsp12-2xStrep-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 ℃.
pLVX-EF1alpha-SARS-CoV-2-nsp12-2xStrep-IRES-Puro vector Sequence
LOCUS pLVX-EF1alpha-SA 11712 bp DNA circular SYN 13-MAY-2021
DEFINITION Lentiviral expression of SARS-CoV-2 protein; transient expression
and generate lentivirus.
ACCESSION .
VERSION .
KEYWORDS pLVX-EF1alpha-SARS-CoV-2-nsp12-2xStrep-IRES-Puro.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11712)
AUTHORS Gordon DE, et al.
TITLE A SARS-CoV-2-Human Protein-Protein Interaction Map Reveals Drug
Targets and Potential Drug Repurposing
JOURNAL BioRxiv March 2020
REFERENCE 2 (bases 1 to 11712)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 11712)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "BioRxiv
March 2020"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..11712
/mol_type="other DNA"
/organism="synthetic DNA construct"
LTR 1..634
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 681..806
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1303..1536
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1721..1765
/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 1914..1955
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
misc_feature 2027..2144
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2338..3519
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
CDS 6367..6390
/label=Strep-Tag II
/note="peptide that binds Strep-Tactin(R), an engineered
form
of streptavidin"
CDS 6427..6450
/label=Strep-Tag II
/note="peptide that binds Strep-Tactin(R), an engineered
form
of streptavidin"
misc_feature 6462..7048
/label=IRES2
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 7068..7664
/label=PuroR
/note="puromycin N-acetyltransferase"
misc_feature 7681..8269
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 8476..9109
/label=5' LTR
/note="5' long terminal repeat (LTR) from HIV-1"
primer_bind complement(9237..9253)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(9237..9253)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(9250..9272)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 9261..9277
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(9285..9315)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(9330..9351)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(9468..9485)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(9639..10227)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(10401..11258)
/label=AmpR
/note="beta-lactamase"
promoter complement(11259..11363)
/label=AmpR promoter
polyA_signal 11411..11545
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
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