Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V015004 | pLV2-TRE3GS-MCS-EGFP-TetOne-IRES-Puro | 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:
- pLV2-TRE3GS-MCS-EGFP-TetOne-IRES-Puro
- Antibiotic Resistance:
- Ampicillin
- Length:
- 10471 bp
- Type:
- Protein expression, Tetracycline inducible
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Selection Marker:
- Puro
- Promoter:
- hPGK
- Growth Temperature:
- 37℃
pLV2-TRE3GS-MCS-EGFP-TetOne-IRES-Puro vector 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.
pLV2-TRE3GS-MCS-EGFP-TetOne-IRES-Puro vector Sequence
LOCUS V015004 10471 bp DNA circular SYN 01-JAN-1980
DEFINITION Exported.
ACCESSION V015004
VERSION V015004
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 10471)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..10471
/mol_type="other DNA"
/organism="synthetic DNA construct"
polyA_signal complement(216..350)
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
promoter 398..502
/label="AmpR promoter"
CDS 503..1360
/label="AmpR"
/note="beta-lactamase"
rep_origin 1534..2119
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 2407..2428
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2443..2473
/label="lac promoter"
/note="promoter for the E. coli lac operon"
LTR complement(2650..3283)
/label="5' LTR"
/note="5' long terminal repeat (LTR) from HIV-1"
misc_feature complement(3490..4078)
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
polyA_signal complement(4092..4316)
/label="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
CDS complement(4344..4940)
/label="PuroR"
/note="puromycin N-acetyltransferase"
misc_feature complement(4960..5546)
/label="IRES2"
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS complement(5562..6305)
/label="Tet-On(R) 3G"
/note="modified rtTA protein that binds tightly to
promoters containing the tet operator in the presence of
doxycycline"
promoter complement(6324..6834)
/label="hPGK promoter"
/note="human phosphoglycerate kinase 1 promoter"
promoter 6851..7218
/label="TRE3GS promoter"
/note="3rd-generation Tet-responsive promoter that can be
activated by binding of Tet-On(R) 3G, modified to eliminate
binding sites for endogenous mammalian transcription
factors"
CDS 7257..7973
/label="EGFP"
/note="enhanced GFP"
polyA_signal complement(8151..8285)
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
misc_feature complement(8328..8445)
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
CDS complement(8517..8558)
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
CDS complement(8707..8751)
/label="gp41 peptide"
/note="antigenic peptide corresponding to amino acids 655
to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
al., 2013)"
misc_feature complement(8936..9169)
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature complement(9666..9791)
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
LTR complement(9838..10471)
/label="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"