Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V017413 | AsCas12a | 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:
- AsCas12a
- Antibiotic Resistance:
- Ampicillin
- Length:
- 12641 bp
- Type:
- Genome editing
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Promoter:
- EM7
- 5' Primer:
- CMV-F
- Growth Temperature:
- 37℃
AsCas12a 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.
AsCas12a vector Sequence
LOCUS V017413 12641 bp DNA circular SYN 01-JAN-1980
DEFINITION Exported.
ACCESSION V017413
VERSION V017413
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 12641)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..12641
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 1..199
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
misc_feature 444..569
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1062..1295
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1480..1524
/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 1673..1714
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 1822..1939
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
misc_RNA 2071..2146
/label="gRNA scaffold"
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
promoter 2208..2419
/label="EF-1-alpha core promoter"
/note="core promoter for human elongation factor
EF-1-alpha"
CDS 2444..6364
/label="AsCpf1"
/note="CRISPR-associated protein from Acidaminococcus sp.
BV3L6 (Zetsche et al., 2015)"
CDS 6413..6436
/label="FLAG"
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
CDS 6446..6502
/label="P2A"
/note="2A peptide from porcine teschovirus-1 polyprotein"
CDS 6503..7096
/label="PuroR"
/note="puromycin N-acetyltransferase"
misc_feature 7115..7703
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 7775..8008
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 8040..8264
/label="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
rep_origin 8310..8738
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 8752..9081
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
promoter 9129..9176
/label="EM7 promoter"
/note="synthetic bacterial promoter"
CDS 9195..9566
/label="BleoR"
/note="antibiotic-binding protein"
polyA_signal 9699..9832
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
promoter complement(9917..9947)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(9962..9983)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 10000..10202
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
rep_origin complement(10271..10859)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(11033..11890)
/label="AmpR"
/note="beta-lactamase"
promoter complement(11891..11995)
/label="AmpR promoter"
enhancer 12261..12640
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"