LOCUS       sequence-141765-         919 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  All-in-One plasmid encoding dual U6 promoter-driven sgRNAs and 
            EGFP-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 919)
  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 919)
  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-GFP
FEATURES             Location/Qualifiers
     source          1..919
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        complement(70..318)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        complement(443..518)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(546..794)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
ORIGIN
        1 acggactagc cttattttaa cttgctattt ctagctctaa aactgagacc ctctcatcga
       61 tggtctcccg gtgtttcgtc ctttccacaa gatatataaa gccaagaaat cgaaatactt
      121 tcaagttacg gtaagcatat gatagtccat tttaaaacat aattttaaaa ctgcaaacta
      181 cccaagaaat tattactttc tacgtcacgt attttgtact aatatctttg tgtttacagt
      241 caaattaatt ctaattatct ctctaacagc cttgtatcgt atatgcaaat atgaaggaat
      301 catgggaaat aggccctctt cctgcccgac ctttcccttt agtgagggtt aatctagagc
      361 catttgtctg cagaattggc gcacgcgcta aaaacggact agccttattt taacttgcta
      421 tttctagctc taaaacaaaa aagcaccgac tcggtgccac tttttcaagt tgataacgga
      481 ctagccttat tttaacttgc tatttctagc tctaaaacag gtcttcgcat ggatccgaag
      541 accccggtgt ttcgtccttt ccacaagata tataaagcca agaaatcgaa atactttcaa
      601 gttacggtaa gcatatgata gtccatttta aaacataatt ttaaaactgc aaactaccca
      661 agaaattatt actttctacg tcacgtattt tgtactaata tctttgtgtt tacagtcaaa
      721 ttaattccaa ttatctctct aacagccttg tatcgtatat gcaaatatga aggaatcatg
      781 ggaaataggc cctcacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
      841 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
      901 ctcaagtcag aggtggcga
//
