Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V017107 | pSLQ7334: pHR-PGK-SV40_NLS-dCas12f-VPR-c- Myc_NLS-mCherry-WPRE | 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:
- pSLQ7334: pHR-PGK-SV40_NLS-dCas12f-VPR-c- Myc_NLS-mCherry-WPRE
- Antibiotic Resistance:
- Ampicillin
- Length:
- 13028 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Promoter:
- mPGK
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
pSLQ7334: pHR-PGK-SV40_NLS-dCas12f-VPR-c- Myc_NLS-mCherry-WPRE 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.
pSLQ7334: pHR-PGK-SV40_NLS-dCas12f-VPR-c- Myc_NLS-mCherry-WPRE vector Sequence
LOCUS V017107 13028 bp DNA circular SYN 01-JAN-1980
DEFINITION Exported.
ACCESSION V017107
VERSION V017107
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 13028)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..13028
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 27..144
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 209..708
/label="PGK promoter"
/note="mouse phosphoglycerate kinase 1 promoter"
CDS 780..800
/label="SV40 NLS"
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
CDS 813..2396
/gene="cas12f"
/label="CRISPR-associated endodeoxyribonuclease Cas12f1"
/note="CRISPR-associated endodeoxyribonuclease Cas12f1 from
Uncultured archaeon. Accession#: A0A482D308"
CDS 2406..2432
/label="HA"
/note="HA (human influenza hemagglutinin) epitope tag"
CDS 3072..3443
/label="RelA (p65) AD"
/note="transcriptional activation domain of human RelA,
also known as p65 (O'Shea and Perkins, 2008)"
CDS 3462..4031
/label="Rta AD"
/note="transcriptional activation domain from the human
herpesvirus 4 (Epstein-Barr virus) replication and
transcription activator Rta/BRLF1 (Hardwick et al., 1992;
Chavez et al., 2015)"
CDS 4059..4085
/label="c-myc NLS"
/note="nuclear localization signal of human c-Myc
proto-oncogene (Dang and Lee, 1988)"
CDS 4098..4802
/label="mCherry"
/note="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
misc_feature 4869..5457
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 5567..5800
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
promoter complement(5825..5843)
/label="SP6 promoter"
/note="promoter for bacteriophage SP6 RNA polymerase"
promoter 6631..6735
/label="AmpR promoter"
CDS 6736..7593
/label="AmpR"
/note="beta-lactamase"
rep_origin 7767..8355
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 8601..8930
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
intron 10064..10129
/label="small t intron"
/note="SV40 (simian virus 40) small t antigen intron"
CDS 10259..10279
/label="SV40 NLS"
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
polyA_signal 10704..10838
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
LTR 11006..11639
/label="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 11686..11811
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 12308..12541
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 12726..12770
/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 12919..12960
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"