LOCUS       sequence-96599-e        1079 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses truncated N-terminal SpCas9 domain fused to a GyrA intein,
            flanked by ITRs for AAV packaging. Combine with C-Terminal Split 
            Cas9 Gyra Intein for full length SpCas9 production.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1079)
  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 1079)
  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 with GyrA intein
FEATURES             Location/Qualifiers
     source          1..1079
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      46..55
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             55..120
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             127..147
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 cctggagcac ctgcctgaaa tcactttttt tcaggttgga ccggtgccac catggactat
       61 aaggaccacg acggagacta caaggatcat gatattgatt acaaagacga tgacgataag
      121 atggccccaa agaagaagcg gaaggtcggt atccacggag tcccagcagc cgacaagaag
      181 tacagcatcg gcctggacat cggcaccaac tctgtgggct gggccgtgat caccgacgag
      241 tacaaggtgc ccagcaagaa attcaaggtg ctgggcaaca ccgaccggca cagcatcaag
      301 aagaacctga tcggagccct gctgttcgac agcggcgaaa cagccgaggc cacccggctg
      361 aagagaaccg ccagaagaag atacaccaga cggaagaacc ggatctgcta tctgcaagag
      421 atcttcagca acgagatggc caaggtggac gacagcttct tccacagact ggaagagtcc
      481 ttcctggtgg aagaggataa gaagcacgag cggcacccca tcttcggcaa catcgtggac
      541 gaggtggcct accacgagaa gtaccccacc atctaccacc tgagaaagaa actggtggac
      601 agcaccgaca aggccgacct gcggctgatc tatctggccc tggcccacat gatcaagttc
      661 cggggccact tcctgatcga gggcgacctg aaccccgaca acagcgacgt ggacaagctg
      721 ttcatccagc tggtgcagac ctacaaccag ctgttcgagg aaaaccccat caacgccagc
      781 ggcgtggacg ccaaggccat cctgtctgcc agactgagca agagcagacg gctggaaaat
      841 ctgatcgccc agctgcccgg cgagaagaag aatggcctgt tcggaaacct gattgccctg
      901 agcctgggcc tgacccccaa cttcaagagc aacttcgacc tggccgagga tgccaaactg
      961 cagctgagca aggacaccta cgacgacgac ctggacaacc tgctggccca gatcggcgac
     1021 cagtacgccg acctgtttct ggccgccaag aacctgtccg acgccatcct gctgagcga
//
