Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V000058 | pfNL43-dE-EGFP | In stock, 1 week for quality controls |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pfNL43-dE-EGFP
- Antibiotic Resistance:
- Ampicillin
- Length:
- 12880 bp
- Type:
- Mammalian Expression, Lentiviral, Retroviral
- Replication origin:
- ori
- Copy Number:
- High Copy
pfNL43-dE-EGFP vector Map
Plasmid 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 ℃.
6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it
General Plasmid Transform Protocol
1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.
2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.
3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.
4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.
5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.
pfNL43-dE-EGFP vector Sequence
LOCUS V000058 12880 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V000058
VERSION V000058
KEYWORDS pfNL43-dE-EGFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 12880)
TITLE NLdDIS
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 12880)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 12880)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..12880
/mol_type="other DNA"
/organism="synthetic DNA construct"
LTR 69..326
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 373..498
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
CDS 482..1981
/label="HIV-1 gag"
/note="gag protein from human immunodeficiency virus 1"
CDS 1777..4785
/label="HIV-1 pol"
/note="pol protein from human immunodeficiency virus 1"
CDS 4733..5308
/gene="vif"
/label="Virion infectivity factor"
/note="Virion infectivity factor from Human
immunodeficiency virus type 1 group M subtype B (isolate
NY5). Accession#: P12504"
CDS 5522..5695
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate BH5). Accession#: P04612"
/label="Protein Tat"
CDS 5753..5995
/gene="vpu"
/label="Protein Vpu"
/note="Protein Vpu from Human immunodeficiency virus type 1
group M subtype B (isolate BH10). Accession#: P69699"
CDS 6066..6782
/label="EGFP"
/note="enhanced GFP"
misc_feature 7304..7537
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 7722..7766
/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 8060..8086
/label="NES"
/note="nuclear export signal from the HIV Rev protein
(Fischer et al., 1995)"
CDS 8332..8358
/gene="nef"
/label="Protein Nef"
/note="Protein Nef from Human immunodeficiency virus type 1
group M subtype D (isolate Z84). Accession#: P12481"
LTR 8621..9254
/label="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"
primer_bind complement(10243..10260)
/label="L4440"
/note="L4440 vector, forward primer"
rep_origin complement(10414..11002)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(11176..12033)
/label="AmpR"
/note="beta-lactamase"
promoter complement(12034..12138)
/label="AmpR promoter"
primer_bind 12206..12224
/label="pBRforEco"
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(12262..12284)
/label="pGEX 3'"
/note="pGEX vectors, reverse primer"
primer_bind 12384..12403
/label="pRS-marker"
/note="pRS vectors, use to sequence yeast selectable
marker"