LOCUS       sequence-257451-         507 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  3xHA tagged Cas9 from S. pyogenes with T2A-EGFP and cloning backbone
            for sgRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 507)
  AUTHORS   Acharya S, Mishra A, Paul D, Ansari AH, Azhar M, Kumar M, Rauthan R,
            Sharma N, Aich M, Sinha D, Sharma S, Jain S, Ray A, Jain S, 
            Ramalingam S, Maiti S, Chakraborty D
  TITLE     Francisella novicida Cas9 interrogates genomic DNA with very high 
            specificity and can be used for mammalian genome editing.
  JOURNAL   Proc Natl Acad Sci U S A. 2019 Oct 15;116(42):20959-20968. doi: 
            10.1073/pnas.1818461116. Epub 2019 Sep 30.
  PUBMED    31570623
REFERENCE   2  (bases 1 to 507)
  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"; doi: 
            "10.1073/pnas.1818461116"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2019-10-15- 15"; volume: "116"; issue: "42"; pages: 
            "20959-20968"
COMMENT     Sequence Label: PX458-3xHA-SpCas9
FEATURES             Location/Qualifiers
     source          1..507
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      7..16
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             22..102
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             109..129
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 accggtgcca ccatgggatc ctacccatac gatgttccag attacgctta cccatacgat
       61 gttccagatt acgcttaccc atacgatgtt ccagattacg ctatggcccc aaagaagaag
      121 cggaaggtcg gtatccacgg agtcccagca gccgacaaga agtacagcat cggcctggac
      181 atcggcacca actctgtggg ctgggccgtg atcaccgacg agtacaaggt gcccagcaag
      241 aaattcaagg tgctgggcaa caccgaccgg cacagcatca agaagaacct gatcggagcc
      301 ctgctgttcg acagcggcga aacagccgag gccacccggc tgaagagaac cgccagaaga
      361 agatacacca gacggaagaa ccggatctgc tatctgcaag agatcttcag caacgagatg
      421 gccaaggtgg acgacagctt cttccacaga ctggaagagt ccttcctggt ggaagaggat
      481 aagaagcacg agcggcaccc catcttc
//
