Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V017113 | pSLQ8703 pHR-R(TRE3G-Fluc-2XcrIFNg-SV40pA)-EF1a-sfGFP-T2A-rtta-WPRE | 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:
- pSLQ8703 pHR-R(TRE3G-Fluc-2XcrIFNg-SV40pA)-EF1a-sfGFP-T2A-rtta-WPRE
- Antibiotic Resistance:
- Ampicillin
- Length:
- 13460 bp
- Type:
- Protein expression, Tetracycline inducible
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Promoter:
- TRE3G
- Growth Strain(s):
- DH5a
- Growth Temperature:
- 37℃
pSLQ8703 pHR-R(TRE3G-Fluc-2XcrIFNg-SV40pA)-EF1a-sfGFP-T2A-rtta-WPRE 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.
pSLQ8703 pHR-R(TRE3G-Fluc-2XcrIFNg-SV40pA)-EF1a-sfGFP-T2A-rtta-WPRE vector Sequence
LOCUS V017113 13460 bp DNA circular SYN 01-JAN-1980
DEFINITION Exported.
ACCESSION V017113
VERSION V017113
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 13460)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..13460
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin complement(463..1051)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(1225..2082)
/label="AmpR"
/note="beta-lactamase"
promoter complement(2083..2187)
/label="AmpR promoter"
promoter 2975..2993
/label="SP6 promoter"
/note="promoter for bacteriophage SP6 RNA polymerase"
LTR complement(3018..3251)
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
CDS 3467..3478
/label="Factor Xa site"
/note="Factor Xa recognition and cleavage site"
CDS complement(4012..4755)
/label="Tet-On(R) 3G"
/note="modified rtTA protein that binds tightly to
promoters containing the tet operator in the presence of
doxycycline"
CDS complement(4831..5544)
/label="superfolder GFP"
/note="GFP variant that folds robustly even when fused to
poorly folded proteins (Nager et al., 2011)"
intron complement(5569..6491)
/label="EF-1-alpha intron A"
/note="intron upstream of the start codon of human
EF-1-alpha"
promoter 6679..7056
/label="TRE3G promoter"
/note="3rd-generation Tet-responsive promoter that can be
activated by binding of Tet-On(R) 3G"
CDS 7066..8715
/label="luciferase"
/note="firefly luciferase"
polyA_signal 8880..9013
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
misc_feature complement(9106..9223)
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
CDS complement(9318..9359)
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
CDS complement(9508..9552)
/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(9737..9970)
/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(10467..10592)
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
LTR complement(10639..11272)
/label="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"
polyA_signal complement(11440..11574)
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
CDS complement(11999..12019)
/label="SV40 NLS"
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
intron complement(12149..12214)
/label="small t intron"
/note="SV40 (simian virus 40) small t antigen intron"