Basic Vector Information
- Vector Name:
- pWPXLd
- Antibiotic Resistance:
- Ampicillin
- Length:
- 10455 bp
- Type:
- Mammalian Expression, Lentiviral
- Replication origin:
- ori
- Copy Number:
- High Copy
- Promoter:
- EF-1α
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- See map
pWPXLd 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 ℃.
pWPXLd vector Sequence
LOCUS pWPXLd. 10455 bp DNA circular SYN 13-MAY-2021
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS pWPXLd.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10455)
TITLE Trono Lab Constitutive Lentiviral Plasmids
REFERENCE 2 (bases 1 to 10455)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 10455)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..10455
/mol_type="other DNA"
/organism="synthetic DNA construct"
LTR 2..635
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 682..807
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1300..1533
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1718..1762
/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 1911..1952
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
promoter 2120..3297
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
misc_feature 3345..3462
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
CDS 3544..4260
/label=EGFP
/note="enhanced GFP"
misc_feature 4340..4928
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 5017..5250
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
promoter complement(5275..5293)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(5583..5602)
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind complement(5816..5835)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 5935..5957
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(5995..6013)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 6081..6185
/label=AmpR promoter
CDS 6186..7043
/label=AmpR
/note="beta-lactamase"
rep_origin 7217..7805
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7959..7976
/label=L4440
/note="L4440 vector, forward primer"
promoter 8051..8380
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
intron 9514..9579
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
CDS 9709..9729
/label=SV40 NLS
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
polyA_signal 10154..10288
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
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