Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V000721 | pLVPT-rtTR-KRAB-2SM2 | 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-rtTR-KRAB-2SM2
- Antibiotic Resistance:
- Ampicillin
- Length:
- 11645 bp
- Type:
- Mammalian Expression, Lentiviral
- Replication origin:
- ori
- Copy Number:
- High Copy
- Promoter:
- mPGK
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- See map
pLVPT-rtTR-KRAB-2SM2 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-rtTR-KRAB-2SM2 vector Sequence
LOCUS V000721 11645 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V000721
VERSION V000721
KEYWORDS pLVPT-rtTR-KRAB-2SM2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 11645)
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 11645)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 11645)
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..11645
/mol_type="other DNA"
/organism="synthetic DNA construct"
LTR 12..645
/label="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 692..817
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1310..1543
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1727..1771
/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 1920..1961
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 2038..2155
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2194..2693
/label="PGK promoter"
/note="mouse phosphoglycerate kinase 1 promoter"
CDS 2750..3466
/label="EGFP"
/note="enhanced GFP"
misc_feature 3549..4111
/label="IRES"
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 4116..4736
/label="TetR"
/note="tetracycline repressor TetR"
CDS 4737..4757
/label="SV40 NLS"
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
CDS 4860..5054
/label="KRAB"
/note="Kruppel-associated box (KRAB) transcriptional
repression domain from the human zinc finger protein ZNF10
(Margolin et al., 1994)"
primer_bind complement(5192..5208)
/label="KS primer"
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5193..5209)
/label="pBluescriptKS"
/note="For pBluescript vector"
misc_feature 5235..5823
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
protein_bind 5961..6231
/label="tetracycline response element"
/note="contains seven copies of the tetracycline operator
tetO"
LTR 6270..6450
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
promoter complement(6475..6493)
/label="SP6 promoter"
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(6783..6802)
/label="pBRrevBam"
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind complement(7016..7035)
/label="pRS-marker"
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 7135..7157
/label="pGEX 3'"
/note="pGEX vectors, reverse primer"
primer_bind complement(7195..7213)
/label="pBRforEco"
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 7281..7385
/label="AmpR promoter"
CDS 7386..8243
/label="AmpR"
/note="beta-lactamase"
rep_origin 8417..9005
/direction=RIGHT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 9159..9176
/label="L4440"
/note="L4440 vector, forward primer"
promoter 9251..9580
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
intron 10714..10779
/label="small t intron"
/note="SV40 (simian virus 40) small t antigen intron"
CDS 10909..10929
/label="SV40 NLS"
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
polyA_signal 11354..11488
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"