LOCUS       sequence-147659-        1062 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses Cas9 codon optimized for Phaeodactylum tricornutum and a 
            sgRNA driven by a U6 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1062)
  AUTHORS   Nymark M, Sharma AK, Sparstad T, Bones AM, Winge P
  TITLE     A CRISPR/Cas9 system adapted for gene editing in marine algae.
  JOURNAL   Sci Rep. 2016 Apr 25;6:24951. doi: 10.1038/srep24951.
  PUBMED    27108533
REFERENCE   2  (bases 1 to 1062)
  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: "2016-04-25"; volume: "6:24951. doi"; pages: " 
            10.1038/srep24951"
COMMENT     Sequence Label: pKS diaCas9_sgRNA
FEATURES             Location/Qualifiers
     source          1..1062
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     6..25
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6..24
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             497..562
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             569..589
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 aattgtaata cgactcacta tagggcgaat tggagctcaa tctcgcctat tcatggtgta
       61 taaaagttca acatccaaag ctagaacttt tggaaagaga aagaatatcc gaatagggca
      121 cggcgtgccg tattgttgga gtggactagc agaaagtgag gaaggcacag gatgagtttt
      181 ctcgagacac atagcttcag cgtcgtgtag gctaggcaga ggtgagtttt ctcgagacat
      241 accttcagcg tcgtcttcac tgtcacagtc aactgacagt aatcgttgat ccggagagat
      301 tcaaaattca atctgtttgg acctggataa gacacaagag cgacatcctg acatgaacgc
      361 cgtaaacagc aaatcctggt tgaacacgta tccttttggg ggcctccgct acgacgctcg
      421 ctccagctgg ggcttcctta ctatacacag cgcgcatatt tcacggttgc cagaagtcaa
      481 gatggcctct agaatggact ataaggacca cgacggagat tacaaggatc atgatattga
      541 ttacaaagac gatgatgata aaatggcccc aaagaagaag cggaaagtcg gtatacacgg
      601 agtcccagca gccgacaaga agtactcgat tggattggac atcggcacca actctgttgg
      661 atgggccgtc atcacggacg agtacaaggt gcccagcaag aaattcaaag tactgggaaa
      721 cactgaccgc cattcgatca agaagaattt gattggagct ttgctgtttg acagcggcga
      781 aacagcggaa gccacgcggc ttaaacgaac cgcgagacga agatatacgc gacggaagaa
      841 ccgtatctgc tatctacaag agattttctc gaatgagatg gcaaaagtcg atgactcttt
      901 ctttcaccga ttggaagaat ccttcctcgt tgaagaggat aagaaacatg agcggcaccc
      961 gatatttgga aacattgtgg acgaggtggc ttaccatgag aaatacccga ctatttacca
     1021 cctgcgaaag aaattggtgg actcgacgga caaagcggat ct
//
