Basic Vector Information
pU6-pegRNA-GG-acceptor 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-pegRNA-GG-acceptor vector Sequence
LOCUS pU6-pegRNA-GG-ac 3004 bp DNA circular SYN 13-MAY-2021 DEFINITION Prime editing in mammalian cells. ACCESSION . VERSION . KEYWORDS pU6-pegRNA-GG-acceptor. SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3004) 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 3004) TITLE Direct Submission REFERENCE 3 (bases 1 to 3004) 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..3004 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 1..241 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" terminator complement(259..290) /label=tonB terminator /note="bidirectional E. coli tonB-P14 transcription terminator" CDS complement(310..984) /label=mRFP1 /note="monomeric derivative of DsRed (Campbell et al., 2002)" RBS 991..1002 /note="strong bacterial ribosome binding site (Elowitz and Leibler, 2000)" protein_bind 1020..1038 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind complement(1045..1063) /label=tet operator /note="bacterial operator O2 for the tetR and tetA genes" rep_origin complement(1174..1762) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(1936..2793) /label=AmpR /note="beta-lactamase" promoter complement(2794..2898) /label=AmpR promoter
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