LOCUS       sequence-141785-         810 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  All-in-One plasmid encoding dual U6 promoter-driven sgRNAs and 
            mCherry-coupled Cas9-D10A nickase to enhance efficient and accurate 
            genome editing.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 810)
  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 810)
  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-mCherry
FEATURES             Location/Qualifiers
     source          1..810
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        complement(75..323)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        complement(448..523)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(551..799)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
ORIGIN
        1 tgataacgga ctagccttat tttaacttgc tatttctagc tctaaaactg agaccctctc
       61 atcgatggtc tcccggtgtt tcgtcctttc cacaagatat ataaagccaa gaaatcgaaa
      121 tactttcaag ttacggtaag catatgatag tccattttaa aacataattt taaaactgca
      181 aactacccaa gaaattatta ctttctacgt cacgtatttt gtactaatat ctttgtgttt
      241 acagtcaaat taattctaat tatctctcta acagccttgt atcgtatatg caaatatgaa
      301 ggaatcatgg gaaataggcc ctcttcctgc ccgacctttc cctttagtga gggttaatct
      361 agagccattt gtctgcagaa ttggcgcacg cgctaaaaac ggactagcct tattttaact
      421 tgctatttct agctctaaaa caaaaaagca ccgactcggt gccacttttt caagttgata
      481 acggactagc cttattttaa cttgctattt ctagctctaa aacaggtctt cgcatggatc
      541 cgaagacccc ggtgtttcgt cctttccaca agatatataa agccaagaaa tcgaaatact
      601 ttcaagttac ggtaagcata tgatagtcca ttttaaaaca taattttaaa actgcaaact
      661 acccaagaaa ttattacttt ctacgtcacg tattttgtac taatatcttt gtgtttacag
      721 tcaaattaat tccaattatc tctctaacag ccttgtatcg tatatgcaaa tatgaaggaa
      781 tcatgggaaa taggccctca catgtgagca
//
