Basic Vector Information
- Vector Name:
- pMA-SpCas9-g1
- Antibiotic Resistance:
- Ampicillin
- Length:
- 2789 bp
- Type:
- Gene knockout
- Replication origin:
- ori
- Host:
- Mammalian cells
- Promoter:
- U6
- Growth Strain(s):
- DH5a
- Growth Temperature:
- 37℃
pMA-SpCas9-g1 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 ℃.
pMA-SpCas9-g1 vector Sequence
LOCUS 62056_16390 2789 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 2789) AUTHORS . TITLE Direct Submission FEATURES Location/Qualifiers source 1..2789 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 42..282 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" misc_RNA 309..384 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" rep_origin complement(681..1269) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(1443..2300) /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(2301..2405) /label=AmpR promoter primer_bind 2721..2737 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 2748..2766 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase"
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