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 Vector Map
Plasmid Resuspension 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 ℃.
AsCas12a vector Sequence
LOCUS 62056_396 12641 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION . VERSION . 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" 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"
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