LOCUS       sequence-155739-         863 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Cas9 nickase optimized for nuclear import..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 863)
  AUTHORS   Shen B, Zhang W, Zhang J, Zhou J, Wang J, Chen L, Wang L, Hodgkins 
            A, Iyer V, Huang X, Skarnes WC
  TITLE     Efficient genome modification by CRISPR-Cas9 nickase with minimal 
            off-target effects.
  JOURNAL   Nat Methods. 2014 Mar 2. doi: 10.1038/nmeth.2857.
  PUBMED    24584192
REFERENCE   2  (bases 1 to 863)
  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: "Nat 
            Methods. 2014 Mar 2. doi: 10.1038/nmeth.2857."
COMMENT     Sequence Label: pST1374-N-NLS-flag-linker-Cas9-D10A
FEATURES             Location/Qualifiers
     source          1..863
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     36..55
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        36..54
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             83..103
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             113..136
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
ORIGIN
        1 actagagaac ccactgctta ctggcttatc gaaattaata cgactcacta tagggagacc
       61 caagctggct agcaccatgg gacctaagaa aaagaggaag gtggcggccg ctgactacaa
      121 ggatgacgac gataaatcta gactggaacc cggcgagaag ccatacaaat gcccagagtg
      181 tggtaaatct ttctctcagt ctggtgctct gaccagacac cagcggaccc acactagaga
      241 caagaaatac tctattggac tggctatcgg gacaaactcc gttggctggg ccgtcataac
      301 cgacgagtat aaggtgccaa gcaagaaatt caaggtgctg ggtaatactg accgccattc
      361 aatcaagaag aacctgatcg gagcactcct cttcgactcc ggtgaaaccg ctgaagctac
      421 tcggctgaag cggaccgcaa ggcggagata cacccgccgc aagaatcgga tatgttatct
      481 gcaagagatc tttagcaacg aaatggctaa ggtggacgac tccttctttc accgcctgga
      541 agagagcttt ctggtggagg aggataagaa acacgagagg caccctatat tcggaaatat
      601 cgtggatgag gtggcttacc atgaaaagta tcctacaatc taccatctga ggaagaagct
      661 ggtggacagc accgataaag cagacctgag gctcatctat ctggccctgg ctcatatgat
      721 aaagtttaga ggacactttc tgatcgaggg cgacctgaat cccgataatt ccgatgtgga
      781 taaactcttc attcaactgg tgcagacata taaccaactg ttcgaggaga atcccataaa
      841 cgcttctggt gtggatgcca agg
//
