Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V014697 | pLenti-Tet-coBxb1-2A-BFP_IRES-iCasp9-2A-Blast_rtTA3 | 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:
- pLenti-Tet-coBxb1-2A-BFP_IRES-iCasp9-2A-Blast_rtTA3
- Antibiotic Resistance:
- Ampicillin
- Length:
- 12668 bp
- Type:
- Protein expression, Tetracycline inducible
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Selection Marker:
- Blast
- Promoter:
- TRE3GV
- Growth Temperature:
- 37℃
pLenti-Tet-coBxb1-2A-BFP_IRES-iCasp9-2A-Blast_rtTA3 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.
pLenti-Tet-coBxb1-2A-BFP_IRES-iCasp9-2A-Blast_rtTA3 vector Sequence
LOCUS V014697 12668 bp DNA circular SYN 01-JAN-1980
DEFINITION Exported.
ACCESSION V014697
VERSION V014697
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 12668)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..12668
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(140..166)
/label="HA"
/note="HA (human influenza hemagglutinin) epitope tag"
CDS complement(167..187)
/label="SV40 NLS"
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
promoter complement(246..595)
/label="TRE3GV promoter"
/note="3rd-generation Tet-responsive promoter that can be
activated by binding of Tet-On(R) 3G, optimized for
retroviral and lentiviral vectors"
misc_feature complement(738..854)
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
CDS complement(931..972)
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
CDS complement(1121..1165)
/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(1350..1583)
/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(2076..2201)
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
LTR complement(2248..2428)
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
promoter complement(2429..2655)
/label="RSV promoter"
/note="Rous sarcoma virus enhancer/promoter"
promoter complement(2683..2701)
/label="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
promoter complement(2770..2800)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(2815..2836)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(3124..3712)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(3886..4743)
/label="AmpR"
/note="beta-lactamase"
promoter complement(4744..4848)
/label="AmpR promoter"
rep_origin complement(4874..5329)
/direction=LEFT
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 5497..5515
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
rep_origin complement(5536..5671)
/direction=LEFT
/label="SV40 ori"
/note="SV40 origin of replication"
polyA_signal complement(5698..5832)
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
LTR complement(5904..6137)
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
CDS complement(6227..6931)
/label="rtTA3"
/note="reverse tetracycline transactivator 3"
promoter complement(7063..7265)
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
enhancer complement(7266..7569)
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
polyA_signal complement(7903..7958)
/label="beta-globin poly(A) signal"
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
CDS complement(8119..8511)
/label="BSD"
/note="blasticidin S deaminase"
CDS complement(8512..8565)
/label="T2A"
/note="2A peptide from Thosea asigna virus capsid protein"
CDS complement(9487..9807)
/label="DmrB"
/note="F36V mutant of FK506-binding protein FKBP12"
misc_feature complement(9817..10403)
/label="IRES2"
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS complement(10528..11235)
/label="mTagBFP2"
/note="enhanced monomeric blue fluorescent protein (Subach
et al., 2011)"