pNL4-3 vector (V004299)

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Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V004299	pNL4-3	In stock, instant shipping	Buy one, get one free!

Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.

Basic Vector Information

Vector Name:: pNL4-3

Antibiotic Resistance:: Ampicillin

Length:: 14825 bp

Type:: HIV-1 vector

Replication origin:: ori

Source/Author:: Adachi A, Gendelman HE, Koenig S, Folks T, Willey R, Rabson A, Martin MA.

Growth Strain(s):: DH5alpha

Growth Temperature:: 37℃

pNL4-3 vector Map

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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.

pNL4-3 vector Sequence

LOCUS       V004299                14825 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004299
VERSION     V004299
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 14825)
  AUTHORS   Adachi A, Gendelman HE, Koenig S, Folks T, Willey R, Rabson A,
            Martin MA.
  TITLE     Production of acquired immunodeficiency syndrome-associated
            retrovirus in human and nonhuman cells transfected with an
            infectious molecular clone
  JOURNAL   J. Virol. 59 (2), 284-291 (1986)
   PUBMED   3016298
REFERENCE   2  (bases 1 to 14825)
  AUTHORS   Bosche WJ, Poon DTK., Ott DE, Hu W-S., Gorelick RJ.
  TITLE     Complete Plasmid Sequence of pNL4-3
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 14825)
  AUTHORS   Bosche WJ, Poon DTK., Ott DE, Hu W-S., Gorelick RJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-NOV-2000) AIDS Vaccine Program, SAIC Frederick and
            Drug Resistance Program, National Cancer Institute at Frederick, PO
            Box B, Frederick, Maryland 21702-1202, USA
REFERENCE   4  (bases 1 to 14825)
  AUTHORS   Strebel KJ, Martin MA.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-MAY-2010) NIAID, Viral Biochemistry, National
            Institutes of Health, Bethesda, MD, USA
REFERENCE   5  (bases 1 to 14825)
  TITLE     Direct Submission
REFERENCE   6  (bases 1 to 14825)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J. Virol.";
            date: "1986"; volume: "59"; issue: "2"; pages: "284-291"
            SGRef: number: 2; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 3; type: "Journal Article"; journalName: "Submitted
            (28-NOV-2000) AIDS Vaccine Program, SAIC Frederick and Drug
            Resistance Program, National Cancer Institute at Frederick, PO Box
            B, Frederick, Maryland 21702-1202, USA"
            SGRef: number: 4; type: "Journal Article"; journalName: "Submitted
            (24-MAY-2010) NIAID, Viral Biochemistry, National Institutes of
            Health, Bethesda, MD, USA"
            SGRef: number: 5; type: "Journal Article"
            On May 24, 2010 this sequence version replaced AF324493.1.
FEATURES             Location/Qualifiers
     source          1..14825
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..634
                     /label="5' long terminal repeat"
                     /note="5' long terminal repeat"
     LTR             377..634
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     repeat_region   454..550
                     /note="R; 5' copy"
     misc_feature    681..806
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     CDS             790..2289
                     /label="HIV-1 gag"
                     /note="gag protein from human immunodeficiency virus 1"
     CDS             2085..5093
                     /label="HIV-1 pol"
                     /note="pol protein from human immunodeficiency virus 1"
     CDS             5041..5616
                     /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             5559..5849
                     /codon_start=1
                     /product="vpr protein"
                     /label="vpr protein"
                     /protein_id="AAK08485.1"
                     /translation="MEQAPEDQGPQREPYNEWTLELLEELKSEAVRHFPRIWLHNLGQH
                     IYETYGDTWAGVEAIIRILQQLLFIHFRIGCRHSRIGVTRQRRARNGASRS"
     misc_feature    5743..5748
                     /label="EcoRI site of recombination"
                     /note="EcoRI site of recombination"
     CDS             5830..6003
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate BH5). Accession#: P04612"
                     /label="Protein Tat"
     CDS             6221..8779
                     /gene="env"
                     /label="Envelope glycoprotein gp160"
                     /note="Envelope glycoprotein gp160 from Human
                     immunodeficiency virus type 1 group M subtype B (isolate
                     MFA). Accession#: P19551"
     CDS             8787..8813
                     /gene="nef"
                     /label="Protein Nef"
                     /note="Protein Nef from Human immunodeficiency virus type 1
                     group M subtype D (isolate Z84). Accession#: P12481"
     LTR             9076..9709
                     /label="3' LTR"
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    9710..11283
                     /label="3' cellular genomic sequence"
                     /note="3' cellular genomic sequence"
     rep_origin      complement(11520..12108)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(12282..13139)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(13140..13244)
                     /label="AmpR promoter"
     misc_feature    13648..14825
                     /label="5' cellular genomic sequence"
                     /note="5' cellular genomic sequence"

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