Basic Vector Information
- Vector Name:
- pMA-SpCas9-g3
- 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-g3 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-g3 vector Sequence
LOCUS 62056_16405 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" misc_RNA complement(264..339) /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" promoter complement(366..606) /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" promoter complement(671..689) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(700..716) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 1032..1136 /label=AmpR promoter CDS 1137..1994 /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 2168..2756 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"
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