LOCUS       sequence-93422-e         740 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses N-terminus of D10A SpCas9 nickase domain fused to a GyrA 
            intein, flanked by ITRs for AAV packaging. Combine with C-Terminal 
            Split Cas9 Gyra Intein for full length SpCas9 nickase production.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 740)
  AUTHORS   Fine EJ, Appleton CM, White DE, Brown MT, Deshmukh H, Kemp ML, Bao G
  TITLE     Trans-spliced Cas9 allows cleavage of HBB and CCR5 genes in human 
            cells using compact expression cassettes.
  JOURNAL   Sci Rep. 2015 Jul 1;5:10777. doi: 10.1038/srep10777.
  PUBMED    26126518
REFERENCE   2  (bases 1 to 740)
  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: "2015-07-1"; volume: "5:10777. doi"; pages: " 
            10.1038/srep10777"
COMMENT     Sequence Label: N-Terminal Split Cas9 D10A Nickase with GyrA i...
FEATURES             Location/Qualifiers
     source          1..740
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     133..152
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        229..304
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        401..686
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 cgatacaagg ctgttagaga gataattgga attaatttga ctgtaaacac aaagatatta
       61 gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta
      121 tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct
      181 ttatatatct tgtggaaagg acgaaacacc gggtcttcga gaagacctgt tttagagcta
      241 gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg
      301 gtgctttttt gttttagagc tagaaatagc aagttaaaat aaggctagtc cgtttttagc
      361 gcgtgcgcca attctgcaga caaatggctc tagaggtacc cgttacataa cttacggtaa
      421 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata gtaacgccaa
      481 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      541 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      601 ccgcctggca ttgtgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      661 acgtattagt catcgctatt accatggtcg aggtgagccc cacgttctgc ttcactctcc
      721 ccatctcccc cccctcccca
//
