Basic Vector Information
pU6-Sp-pegRNA-HEK3_CTT_ins 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 ℃.
pU6-Sp-pegRNA-HEK3_CTT_ins vector Sequence
LOCUS 40924_44659 2305 bp DNA circular SYN 13-MAY-2021 DEFINITION S. pyogenes pegRNA for CTT insertion at the HEK3 site of human cells using prime editing. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2305) AUTHORS Anzalone AV, Randolph PB, Davis JR, Sousa AA, Koblan LW, Levy JM, Chen PJ, Wilson C, Newby GA, Raguram A, Liu DR TITLE Search-and-replace genome editing without double-strand breaks or donor DNA. JOURNAL Nature. 2019 Oct 21. pii: 10.1038/s41586-019-1711-4. doi: 10.1038/s41586-019-1711-4. PUBMED 31634902 REFERENCE 2 (bases 1 to 2305) TITLE Direct Submission REFERENCE 3 (bases 1 to 2305) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nature. 2019 Oct 21. pii: 10.1038/s41586-019-1711-4. doi: 10.1038/s41586-019-1711-4." COMMENT SGRef: number: 2; type: "Journal Article" FEATURES Location/Qualifiers source 1..2305 /mol_type="other DNA" /organism="synthetic DNA construct" misc_RNA 29..104 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" rep_origin complement(234..822) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(996..1853) /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(1854..1958) /label=AmpR promoter promoter 2065..2305 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA"
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