LOCUS       sequence-145605-         823 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  All-in-One plasmid encoding dual U6 promoter-driven sgRNAs and 
            Cas9-D10A nickase linked via 2A peptide with puromycin resistant 
            marker to enhance efficient and accurate genome editing.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 823)
  AUTHORS   Chiang TW, le Sage C, Larrieu D, Demir M, Jackson SP
  TITLE     CRISPR-Cas9(D10A) nickase-based genotypic and phenotypic screening 
            to enhance genome editing.
  JOURNAL   Sci Rep. 2016 Apr 15;6:24356. doi: 10.1038/srep24356.
  PUBMED    27079678
REFERENCE   2  (bases 1 to 823)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep."; 
            date: "2016-04-15"; volume: "6:24356. doi"; pages: " 
            10.1038/srep24356"
COMMENT     Sequence Label: AIO-Puro
FEATURES             Location/Qualifiers
     source          1..823
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             271..318
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             334..390
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
ORIGIN
        1 agagtgatcc tggccgacgc taatctggac aaagtgctgt ccgcctacaa caagcaccgg
       61 gataagccca tcagagagca ggccgagaat atcatccacc tgtttaccct gaccaatctg
      121 ggagcccctg ccgccttcaa gtactttgac accaccatcg accggaagag gtacaccagc
      181 accaaagagg tgctggacgc caccctgatc caccagagca tcaccggcct gtacgagaca
      241 cggatcgacc tgtctcagct gggaggcgac aaaaggccgg cggccacgaa aaaggccggc
      301 caggcaaaaa agaaaaagga attcggctcc ggagcaacaa acttctctct gctgaaacaa
      361 gccggagatg tcgaagagaa tcctggaccg accgagtaca agcccacggt gcgcctcgcc
      421 acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc
      481 gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa
      541 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc
      601 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc
      661 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc
      721 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggagtctcg
      781 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tcc
//
