Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V013874 | pLKO-H1-2O2-shRNA-hPGK-TetR-PGK-Puro | 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:
- pLKO-H1-2O2-shRNA-hPGK-TetR-PGK-Puro
- Antibiotic Resistance:
- Ampicillin
- Length:
- 11100 bp
- Type:
- RNA interference, Tetracycline inducible
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Selection Marker:
- Puro
- Promoter:
- hPGK
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
pLKO-H1-2O2-shRNA-hPGK-TetR-PGK-Puro 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.
pLKO-H1-2O2-shRNA-hPGK-TetR-PGK-Puro vector Sequence
LOCUS V013874 11100 bp DNA circular SYN 01-JAN-1980
DEFINITION Exported.
ACCESSION V013874
VERSION V013874
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 11100)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..11100
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 7..507
/label="hPGK promoter"
/note="human phosphoglycerate kinase 1 promoter"
intron 516..1088
/label="beta-globin intron"
/note="intron from rabbit beta-globin gene"
promoter 1143..1161
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 1172..1792
/label="TetR"
/note="tetracycline repressor TetR"
promoter 1821..2320
/label="PGK promoter"
/note="mouse phosphoglycerate kinase 1 promoter"
CDS 2341..2937
/label="PuroR"
/note="puromycin N-acetyltransferase"
misc_feature 2954..3542
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 3614..3847
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 3919..4053
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin 4080..4215
/label="SV40 ori"
/note="SV40 origin of replication"
promoter complement(4236..4254)
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
rep_origin 4422..4877
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 4903..5007
/label="AmpR promoter"
CDS 5008..5865
/label="AmpR"
/note="beta-lactamase"
rep_origin 6039..6627
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 6915..6936
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 6951..6981
/label="lac promoter"
/note="promoter for the E. coli lac operon"
promoter 7050..7068
/label="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 7096..7322
/label="RSV promoter"
/note="Rous sarcoma virus enhancer/promoter"
LTR 7323..7503
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 7550..7675
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 8168..8401
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 8586..8630
/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 8779..8820
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
promoter 8919..9013
/label="H1-2O2 promoter"
/note="doxycycline-inducible variant of the human H1 RNA
promoter (Henriksen et al., 2007)"
promoter 10000..10507
/label="hPGK promoter"
/note="human phosphoglycerate kinase 1 promoter"
misc_feature 10931..11048
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"