LOCUS       sequence-100854-         351 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses D10A mutant (nickase) of human codon-optimized Cas9 
            (derived from Streptococcus pyogenes) and pruomycin resistance 
            gene..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 351)
  AUTHORS   Li HL, Fujimoto N, Sasakawa N, Shirai S, Ohkame T, Sakuma T, Tanaka 
            M, Amano N, Watanabe A, Sakurai H, Yamamoto T, Yamanaka S, Hotta A
  TITLE     Precise Correction of the Dystrophin Gene in Duchenne Muscular 
            Dystrophy Patient Induced Pluripotent Stem Cells by TALEN and 
            CRISPR-Cas9.
  JOURNAL   Stem Cell Reports. 2014 Nov 25. pii: S2213-6711(14)00335-X. doi: 
            10.1016/j.stemcr.2014.10.013.
  PUBMED    25434822
REFERENCE   2  (bases 1 to 351)
  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: "Stem Cell 
            Reports. 2014 Nov 25. pii: S2213-6711(14)00335-X. doi: 
            10.1016/j.stemcr.2014.10.013."
COMMENT     Sequence Label: pHL-EF1a-SphcCas9(D10A)-iP-A
FEATURES             Location/Qualifiers
     source          1..351
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    171..195
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(208..228)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     RBS             320..328
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
ORIGIN
        1 ccaggtttcc gggccctcac attgccaaaa gacggcaata tggtggaaaa taacatatag
       61 acaaacgcac accggcctta ttccaagcgg cttcggccag taacgttagg ggggggggag
      121 ggagaggggg gggggcggaa ttcgattcga ggccgcgttt tagctagatc accactttgt
      181 acaagaaagc tgggtctaga ggtctcacac cttcctcttc ttcttggggt cagctccgga
      241 gtcgccgccc agctgagaca ggtcaatcct tgtttcatac aggccagtaa tgctctgatg
      301 aatcagggta gcgtccagca cctccttagt gcttgtgtat cgtttgcgat c
//
