LOCUS       sequence-68915-e         938 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expesses the human codon usage Cas9 nuclease (Mali et al. Science 
            339, 823-826, 2013) with an N-terminal GFP tag from the 35S promoter
            in the plant tissue .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 938)
  AUTHORS   Nekrasov V, Staskawicz B, Weigel D, Jones JD, Kamoun S
  TITLE     Targeted mutagenesis in the model plant Nicotiana benthamiana using 
            Cas9 RNA-guided endonuclease.
  JOURNAL   Nat Biotechnol. 2013 Aug 8;31(8):691-3. doi: 10.1038/nbt.2655.
  PUBMED    23929340
REFERENCE   2  (bases 1 to 938)
  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: "10.1038/nbt"; 
            journalName: "Nat Biotechnol"; date: "2013-08-8- 8"; volume: "31"; 
            issue: "8"; pages: "691-3"
COMMENT     Sequence Label: pK7WGF2::hCas9
FEATURES             Location/Qualifiers
     source          1..938
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             join(107..109,110..112,113..823)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(107..109,110..112,113..823)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(152..173)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(413..432)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     760..781
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    830..854
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 atttggagag gactccggta tttttacaac aattaccaca acaaaacaaa caacaaacaa
       61 cattacaatt tactattcta gtcgacctgc aggcggccgc actagtatgg tgagcaaggg
      121 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
      181 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
      241 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
      301 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
      361 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
      421 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
      481 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
      541 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
      601 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
      661 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
      721 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
      781 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aaggatatca caagtttgta
      841 caaaaaagca ggctccgcgg ccgccccctt caccatggac aagaagtact ccattgggct
      901 cgatatcggc acaaacagcg tcggctgggc cgtcatta
//
