Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V013101 | 041 pLVPTH2-tTR-KRAB-Cerulean | 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:
- 041 pLVPTH2-tTR-KRAB-Cerulean
- Antibiotic Resistance:
- Ampicillin
- Length:
- 12015 bp
- Type:
- RNA interference
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Promoter:
- mPGK
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
041 pLVPTH2-tTR-KRAB-Cerulean 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.
041 pLVPTH2-tTR-KRAB-Cerulean vector Sequence
LOCUS V013101 12015 bp DNA circular SYN 01-JAN-1980
DEFINITION Exported.
ACCESSION V013101
VERSION V013101
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 12015)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..12015
/mol_type="other DNA"
/organism="synthetic DNA construct"
LTR 1..634
/label="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 681..806
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1299..1532
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1717..1761
/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 1910..1951
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 2028..2145
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2184..2683
/label="PGK promoter"
/note="mouse phosphoglycerate kinase 1 promoter"
CDS 2746..3462
/label="mCerulean"
/note="enhanced monomeric variant of CFP (Rizzo et al.,
2004)"
protein_bind complement(3467..3491)
/label="attB2"
/note="recombination site for the Gateway(R) BP reaction"
misc_feature 3512..4074
/label="IRES"
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 4079..4699
/label="TetR"
/note="tetracycline repressor TetR"
CDS 4700..4720
/label="SV40 NLS"
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
CDS 4823..5017
/label="KRAB"
/note="Kruppel-associated box (KRAB) transcriptional
repression domain from the human zinc finger protein ZNF10
(Margolin et al., 1994)"
misc_feature 5187..5775
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
protein_bind 5913..6183
/label="tetracycline response element"
/note="contains seven copies of the tetracycline operator
tetO"
promoter 6374..6586
/label="H1 promoter"
/note="human H1 RNA promoter"
LTR 6629..6809
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
promoter complement(6834..6852)
/label="SP6 promoter"
/note="promoter for bacteriophage SP6 RNA polymerase"
promoter 7640..7744
/label="AmpR promoter"
CDS 7745..8602
/label="AmpR"
/note="beta-lactamase"
rep_origin 8776..9364
/direction=RIGHT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 9610..9939
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
intron 11073..11138
/label="small t intron"
/note="SV40 (simian virus 40) small t antigen intron"
CDS 11268..11288
/label="SV40 NLS"
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
polyA_signal 11713..11847
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"