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SG3ΔENV IN D116A vector (V017068)

Price Information

Cat No. Plasmid Name Availability Add to cart
V017068 SG3ΔENV IN D116A 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:
SG3ΔENV IN D116A
Antibiotic Resistance:
Ampicillin
Length:
14788 bp
Type:
Protein expression
Replication origin:
ori
Host:
Mammalian cells
Promoter:
T7
Growth Strain(s):
Stbl3
Growth Temperature:
37℃

SG3ΔENV IN D116A vector Map

Copy Sequence View Map
SG3ΔENV IN D116A14788 bp700140021002800350042004900560063007000770084009100980010500112001190012600133001400014700HIV-1 polRREgp41 peptideProtein Tat3' LTRT7 promoterlac promoterCAP binding siteoriAmpRAmpR promoterf1 oriM13 fwd3' LTRHIV-1 Psi

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.

SG3ΔENV IN D116A vector Sequence

LOCUS       V017068                14788 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V017068
VERSION     V017068
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 14788)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..14788
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1232..4240
                     /label="HIV-1 pol"
                     /note="pol protein from human immunodeficiency virus 1"
     misc_feature    6928..7161
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             7346..7390
                     /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             7539..7580
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     LTR             8245..8878
                     /label="3' LTR"
                     /note="3' long terminal repeat (LTR) from HIV-1"
     promoter        complement(9368..9386)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(9453..9483)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(9498..9519)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(9807..10395)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(10569..11426)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(11427..11531)
                     /label="AmpR promoter"
     rep_origin      11558..12013
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     12154..12170
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     LTR             13936..14569
                     /label="3' LTR"
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    14616..14741
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     CDS             join(14725..14788,1..1436)
                     /gene="gag"
                     /label="Gag polyprotein"
                     /note="Gag polyprotein from Human immunodeficiency virus
                     type 1 group M subtype B (strain 89.6). Accession#: Q73367"