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pNL4-3-HA-R-E- vector (V014776)

Price Information

Cat No. Plasmid Name Availability Add to cart
V014776 pNL4-3-HA-R-E- In stock, 1 week for quality controls

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

Basic Vector Information

Vector Name:
pNL4-3-HA-R-E-
Antibiotic Resistance:
Ampicillin
Length:
14609 bp
Type:
Protein expression
Replication origin:
ori
Host:
Mammalian cells, Lentivirus
Growth Temperature:
37℃

pNL4-3-HA-R-E- vector Map

Copy Sequence View Map
pNL4-3-HA-R-E-14609 bp70014002100280035004200490056006300700077008400910098001050011200119001260013300140005' LTR (truncated)HIV-1 PsiHIV-1 polProtein TatProtein VpuRREgp41 peptideNESHA5' LTR (truncated)oriAmpRAmpR promoter

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-HA-R-E- vector Sequence

LOCUS       V014776                14609 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014776
VERSION     V014776
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 14609)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..14609
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1290..1547
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1594..1719
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     CDS             1703..3202
                     /label="HIV-1 gag"
                     /note="gag protein from human immunodeficiency virus 1"
     CDS             2998..6006
                     /label="HIV-1 pol"
                     /note="pol protein from human immunodeficiency virus 1"
     CDS             5954..6529
                     /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             6743..6916
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate BH5). Accession#: P04612"
                     /label="Protein Tat"
     CDS             6974..7216
                     /gene="vpu"
                     /label="Protein Vpu"
                     /note="Protein Vpu from Human immunodeficiency virus type 1
                     group M subtype B (isolate BH10). Accession#: P69699"
     misc_feature    8672..8905
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             9090..9134
                     /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             9428..9454
                     /label="NES"
                     /note="nuclear export signal from the HIV Rev protein
                     (Fischer et al., 1995)"
     CDS             9554..9580
                     /label="HA"
                     /note="HA (human influenza hemagglutinin) epitope tag"
     LTR             10149..10406
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(12217..12805)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(12979..13836)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(13837..13941)
                     /label="AmpR promoter"