Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V006525 | pLVPT-tTR-KRAB | 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:
- pLVPT-tTR-KRAB
- Antibiotic Resistance:
- Ampicillin
- Length:
- 11634 bp
- Type:
- Mammalian Expression, Lentiviral
- Replication origin:
- ori
- Copy Number:
- High Copy
- Promoter:
- mPGK
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- See map
pLVPT-tTR-KRAB 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.
pLVPT-tTR-KRAB vector Sequence
LOCUS V006525 11634 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V006525
VERSION V006525
KEYWORDS pLVPT-tTR-KRAB
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 11634)
AUTHORS Szulc J, Wiznerowicz M, Sauvain MO, Trono D, Aebischer P
TITLE A versatile tool for conditional gene expression and knockdown.
JOURNAL Nat Methods. 2006 Feb . 3(2):109-16.
PUBMED 16432520
REFERENCE 2 (bases 1 to 11634)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 11634)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat
Methods. 2006 Feb . 3(2):109-16."
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..11634
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 176..217
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 294..411
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 450..949
/label="PGK promoter"
/note="mouse phosphoglycerate kinase 1 promoter"
CDS 1006..1722
/label="EGFP"
/note="enhanced GFP"
misc_feature 1805..2367
/label="IRES"
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 2372..2992
/label="TetR"
/note="tetracycline repressor TetR"
CDS 2993..3013
/label="SV40 NLS"
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
CDS 3116..3310
/label="KRAB"
/note="Kruppel-associated box (KRAB) transcriptional
repression domain from the human zinc finger protein ZNF10
(Margolin et al., 1994)"
misc_feature 3480..4068
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
protein_bind 4206..4476
/label="tetracycline response element"
/note="contains seven copies of the tetracycline operator
tetO"
LTR 4515..4695
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
promoter complement(4720..4738)
/label="SP6 promoter"
/note="promoter for bacteriophage SP6 RNA polymerase"
promoter 5526..5630
/label="AmpR promoter"
CDS 5631..6488
/label="AmpR"
/note="beta-lactamase"
rep_origin 6662..7250
/direction=RIGHT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 7496..7825
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
intron 8959..9024
/label="small t intron"
/note="SV40 (simian virus 40) small t antigen intron"
CDS 9154..9174
/label="SV40 NLS"
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
polyA_signal 9599..9733
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
LTR 10277..10534
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 10581..10706
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 11199..11432
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS join(11617..11634,1..27)
/label="gp41 peptide"
/note="antigenic peptide corresponding to amino acids 655
to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
al., 2013)"