Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V012797 | pNL4-3.Luc.R-E- | In stock (lyophilized plasmid) |
Buy one, get one free! |
Two vials of lyophilized plasmid will be delivered, each vial is about 5µg.
Basic Vector Information
Firefly luciferase gene was inserted into the pNL4-3 nef gene. Two frameshifts (5'Env and Vpr aa 26) render this clone Env- and Vpr-. Competent for a single round of replication. Requires cotransfection with env expression vector to produce infectious virus. Virus can be produced by transfecting 2x10^6 293 or 293T cells with 20 ug of the NL4-3 DNA, or with 10 ug NL4-3 DNA and 10 ug env expression vector DNA. Transfections can be performed in a 10 cm^2 tissue culture dish using standard calcium phosphate protocols. Virus is typically harvested 48 hours post-transfection. Infections should be performed in a total volume of 0.5 ml. Amphotropic pseudotypes generally have much higher infectivity than those bearing HIV-1 env. Cultures infected with the luciferase viruses can be lysed 2-5 days post-infection and assayed using commercial lysis buffer and luciferase reagents
- Vector Name:
- pNL4-3.Luc.R-E-
- Antibiotic Resistance:
- Ampicillin
- Length:
- 16390 bp
- Type:
- Viral Expression & Packaging Vectors
- Replication origin:
- ori
- Source/Author:
- Dr. Nathaniel Landau, Aaron Diamond AIDS Research Center, The Rockefeller University.
- Copy Number:
- High copy number
- Expression Method:
- Constiutive, Stable
pNL4-3.Luc.R-E- 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 ℃.
pNL4-3.Luc.R-E- vector Sequence
LOCUS pnl4-3-luc-r-e-. 16390 bp DNA circular SYN 08-OCT-2021
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS pnl4-3-luc-r-e-.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 16390)
AUTHORS Triple Threat
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..16390
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 669..773
/label=AmpR promoter
CDS 774..1631
/label=AmpR
/note="beta-lactamase"
rep_origin 1805..2393
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
LTR complement(4204..4837)
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
CDS complement(5027..6673)
/label=luciferase
/note="firefly luciferase"
CDS complement(6699..9257)
/gene="env"
/label=env
/note="Envelope glycoprotein gp160 from Human
immunodeficiency virus type 1 group M subtype B (isolate
MFA). Accession#: P19551"
CDS complement(9475..9648)
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate BH5). Accession#: P04612"
CDS complement(9862..10437)
/gene="vif"
/label=vif
/note="Virion infectivity factor from Human
immunodeficiency virus type 1 group M subtype B (isolate
NY5). Accession#: P12504"
CDS complement(10385..13393)
/label=HIV-1 pol
/note="pol protein from human immunodeficiency virus 1"
CDS complement(13189..14688)
/label=HIV-1 gag
/note="gag protein from human immunodeficiency virus 1"
misc_feature complement(14672..14797)
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
LTR complement(14844..15101)
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"