LOCUS       sequence-332335-        2082 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  SpCas9-NG-mediated genome editing..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 2082)
  AUTHORS   Oura S, Noda T, Morimura N, Hitoshi S, Nishimasu H, Nagai Y, Nureki 
            O, Ikawa M
  TITLE     Precise CAG repeat contraction in a Huntington's Disease mouse model
            is enabled by gene editing with SpCas9-NG.
  JOURNAL   Commun Biol. 2021 Jun 23;4(1):771. doi: 10.1038/s42003-021-02304-w.
  PUBMED    34163001
REFERENCE   2  (bases 1 to 2082)
  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: "Commun 
            Biol."; date: "2021-06-23"; volume: "4(1):771. doi"; pages: " 
            10.1038/s42003-021-02304-w"
COMMENT     Sequence Label: pX459-SpCas9-NG
FEATURES             Location/Qualifiers
     source          1..2082
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      1..10
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             10..75
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             82..102
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             127..1842
                     /codon_start=1
                     /product="N-terminal portion of Streptococcus pyogenes Cas9
                     (Zetsche et al., 2015)"
                     /label=Cas9(N)
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIE"
ORIGIN
        1 gccaccatgg actataagga ccacgacgga gactacaagg atcatgatat tgattacaaa
       61 gacgatgacg ataagatggc cccaaagaag aagcggaagg tcggtatcca cggagtccca
      121 gcagccgaca agaagtacag catcggcctg gacatcggca ccaactctgt gggctgggcc
      181 gtgatcaccg acgagtacaa ggtgcccagc aagaaattca aggtgctggg caacaccgac
      241 cggcacagca tcaagaagaa cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc
      301 gaggccaccc ggctgaagag aaccgccaga agaagataca ccagacggaa gaaccggatc
      361 tgctatctgc aagagatctt cagcaacgag atggccaagg tggacgacag cttcttccac
      421 agactggaag agtccttcct ggtggaagag gataagaagc acgagcggca ccccatcttc
      481 ggcaacatcg tggacgaggt ggcctaccac gagaagtacc ccaccatcta ccacctgaga
      541 aagaaactgg tggacagcac cgacaaggcc gacctgcggc tgatctatct ggccctggcc
      601 cacatgatca agttccgggg ccacttcctg atcgagggcg acctgaaccc cgacaacagc
      661 gacgtggaca agctgttcat ccagctggtg cagacctaca accagctgtt cgaggaaaac
      721 cccatcaacg ccagcggcgt ggacgccaag gccatcctgt ctgccagact gagcaagagc
      781 agacggctgg aaaatctgat cgcccagctg cccggcgaga agaagaatgg cctgttcgga
      841 aacctgattg ccctgagcct gggcctgacc cccaacttca agagcaactt cgacctggcc
      901 gaggatgcca aactgcagct gagcaaggac acctacgacg acgacctgga caacctgctg
      961 gcccagatcg gcgaccagta cgccgacctg tttctggccg ccaagaacct gtccgacgcc
     1021 atcctgctga gcgacatcct gagagtgaac accgagatca ccaaggcccc cctgagcgcc
     1081 tctatgatca agagatacga cgagcaccac caggacctga ccctgctgaa agctctcgtg
     1141 cggcagcagc tgcctgagaa gtacaaagag attttcttcg accagagcaa gaacggctac
     1201 gccggctaca ttgacggcgg agccagccag gaagagttct acaagttcat caagcccatc
     1261 ctggaaaaga tggacggcac cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg
     1321 cggaagcagc ggaccttcga caacggcagc atcccccacc agatccacct gggagagctg
     1381 cacgccattc tgcggcggca ggaagatttt tacccattcc tgaaggacaa ccgggaaaag
     1441 atcgagaaga tcctgacctt ccgcatcccc tactacgtgg gccctctggc caggggaaac
     1501 agcagattcg cctggatgac cagaaagagc gaggaaacca tcaccccctg gaacttcgag
     1561 gaagtggtgg acaagggcgc ttccgcccag agcttcatcg agcggatgac caacttcgat
     1621 aagaacctgc ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga gtacttcacc
     1681 gtgtataacg agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc
     1741 ctgagcggcg agcagaaaaa ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg
     1801 accgtgaagc agctgaaaga ggactacttc aagaaaatcg agtgcttcga ctccgtggaa
     1861 atctccggcg tggaagatcg gttcaacgcc tccctgggca cataccacga tctgctgaaa
     1921 attatcaagg acaaggactt cctggacaat gaggaaaacg aggacattct ggaagatatc
     1981 gtgctgaccc tgacactgtt tgaggacaga gagatgatcg aggaacggct gaaaacctat
     2041 gcccacctgt tcgacgacaa agtgatgaag cagctgaagc gg
//
