LOCUS       sequence-73965-e        3345 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Mammalian N. meningitidis Cas9 expression, human optimized.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3345)
  AUTHORS   Esvelt KM, Mali P, Braff JL, Moosburner M, Yaung SJ, Church GM
  TITLE     Orthogonal Cas9 proteins for RNA-guided gene regulation and editing.
  JOURNAL   Nat Methods. 2013 Sep 29. doi: 10.1038/nmeth.2681.
  PUBMED    24076762
REFERENCE   2  (bases 1 to 3345)
  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. 2013 Sep 29. doi: 10.1038/nmeth.2681."
COMMENT     Sequence Label: M-NMcas
FEATURES             Location/Qualifiers
     source          1..3345
                     /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             7..3252
                     /codon_start=1
                     /product="Cas9 endonuclease from the Neisseria meningitidis
                     Type II CRISPR/Cas system (Esvelt et al., 2013)"
                     /label=NmCas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MAAFKPNPINYILGLDIGIASVGWAMVEIDEDENPICLIDLGVRV
                     FERAEVPKTGDSLAMARRLARSVRRLTRRRAHRLLRARRLLKREGVLQAADFDENGLIK
                     SLPNTPWQLRAAALDRKLTPLEWSAVLLHLIKHRGYLSQRKNEGETADKELGALLKGVA
                     DNAHALQTGDFRTPAELALNKFEKESGHIRNQRGDYSHTFSRKDLQAELILLFEKQKEF
                     GNPHVSGGLKEGIETLLMTQRPALSGDAVQKMLGHCTFEPAEPKAAKNTYTAERFIWLT
                     KLNNLRILEQGSERPLTDTERATLMDEPYRKSKLTYAQARKLLGLEDTAFFKGLRYGKD
                     NAEASTLMEMKAYHAISRALEKEGLKDKKSPLNLSPELQDEIGTAFSLFKTDEDITGRL
                     KDRIQPEILEALLKHISFDKFVQISLKALRRIVPLMEQGKRYDEACAEIYGDHYGKKNT
                     EEKIYLPPIPADEIRNPVVLRALSQARKVINGVVRRYGSPARIHIETAREVGKSFKDRK
                     EIEKRQEENRKDREKAAAKFREYFPNFVGEPKSKDILKLRLYEQQHGKCLYSGKEINLG
                     RLNEKGYVEIDHALPFSRTWDDSFNNKVLVLGSENQNKGNQTPYEYFNGKDNSREWQEF
                     KARVETSRFPRSKKQRILLQKFDEDGFKERNLNDTRYVNRFLCQFVADRMRLTGKGKKR
                     VFASNGQITNLLRGFWGLRKVRAENDRHHALDAVVVACSTVAMQQKITRFVRYKEMNAF
                     DGKTIDKETGEVLHQKTHFPQPWEFFAQEVMIRVFGKPDGKPEFEEADTPEKLRTLLAE
                     KLSSRPEAVHEYVTPLFVSRAPNRKMSGQGHMETVKSAKRLDEGVSVLRVPLTQLKLKD
                     LEKMVNREREPKLYEALKARLEAHKDDPAKAFAEPFYKYDKAGNRTQQVKAVRVEQVQK
                     TGVWVRNHNGIADNATMVRVDVFEKGDKYYLVPIYSWQVAKGILPDRAVVQGKDEEDWQ
                     LIDDSFNFKFSLHPNDLVEVITKKARMFGYFASCHRGTGNINIRIHDLDHKIGKNGILE
                     GIGVKTALSFQKYQIDELGKEIRPCRLKKRPPVR"
     regulatory      2839..2848
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3265..3285
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3289..3309
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3313..3333
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 gccaccatgg ccgccttcaa gcccaacccc atcaactaca tcctgggcct ggacatcggc
       61 atcgccagcg tgggctgggc catggtggag atcgacgagg acgagaaccc catctgcctg
      121 atcgacctgg gtgtgcgcgt gttcgagcgc gctgaggtgc ccaagactgg tgacagtctg
      181 gctatggctc gccggcttgc tcgctctgtt cggcgcctta ctcgccggcg cgctcaccgc
      241 cttctgcgcg ctcgccgcct gctgaagcgc gagggtgtgc tgcaggctgc cgacttcgac
      301 gagaacggcc tgatcaagag cctgcccaac actccttggc agctgcgcgc tgccgctctg
      361 gaccgcaagc tgactcctct ggagtggagc gccgtgctgc tgcacctgat caagcaccgc
      421 ggctacctga gccagcgcaa gaacgagggc gagaccgccg acaaggagct gggtgctctg
      481 ctgaagggcg tggccgacaa cgcccacgcc ctgcagactg gtgacttccg cactcctgct
      541 gagctggccc tgaacaagtt cgagaaggag agcggccaca tccgcaacca gcgcggcgac
      601 tacagccaca ccttcagccg caaggacctg caggccgagc tgatcctgct gttcgagaag
      661 cagaaggagt tcggcaaccc ccacgtgagc ggcggcctga aggagggcat cgagaccctg
      721 ctgatgaccc agcgccccgc cctgagcggc gacgccgtgc agaagatgct gggccactgc
      781 accttcgagc cagccgagcc caaggccgcc aagaacacct acaccgccga gcgcttcatc
      841 tggctgacca agctgaacaa cctgcgcatc ctggagcagg gcagcgagcg ccccctgacc
      901 gacaccgagc gcgccaccct gatggacgag ccctaccgca agagcaagct gacctacgcc
      961 caggcccgca agctgctggg tctggaggac accgccttct tcaagggcct gcgctacggc
     1021 aaggacaacg ccgaggccag caccctgatg gagatgaagg cctaccacgc catcagccgc
     1081 gccctggaga aggagggcct gaaggacaag aagagtcctc tgaacctgag ccccgagctg
     1141 caggacgaga tcggcaccgc cttcagcctg ttcaagaccg acgaggacat caccggccgc
     1201 ctgaaggacc gcatccagcc cgagatcctg gaggccctgc tgaagcacat cagcttcgac
     1261 aagttcgtgc agatcagcct gaaggccctg cgccgcatcg tgcccctgat ggagcagggc
     1321 aagcgctacg acgaggcctg cgccgagatc tacggcgacc actacggcaa gaagaacacc
     1381 gaggagaaga tctacctgcc tcctatcccc gccgacgaga tccgcaaccc cgtggtgctg
     1441 cgcgccctga gccaggcccg caaggtgatc aacggcgtgg tgcgccgcta cggcagcccc
     1501 gcccgcatcc acatcgagac cgcccgcgag gtgggcaaga gcttcaagga ccgcaaggag
     1561 atcgagaagc gccaggagga gaaccgcaag gaccgcgaga aggccgccgc caagttccgc
     1621 gagtacttcc ccaacttcgt gggcgagccc aagagcaagg acatcctgaa gctgcgcctg
     1681 tacgagcagc agcacggcaa gtgcctgtac agcggcaagg agatcaacct gggccgcctg
     1741 aacgagaagg gctacgtgga gatcgaccac gccctgccct tcagccgcac ctgggacgac
     1801 agcttcaaca acaaggtgct ggtgctgggc agcgagaacc agaacaaggg caaccagacc
     1861 ccctacgagt acttcaacgg caaggacaac agccgcgagt ggcaggagtt caaggcccgc
     1921 gtggagacca gccgcttccc ccgcagcaag aagcagcgca tcctgctgca gaagttcgac
     1981 gaggacggct tcaaggagcg caacctgaac gacacccgct acgtgaaccg cttcctgtgc
     2041 cagttcgtgg ccgaccgcat gcgcctgacc ggcaagggca agaagcgcgt gttcgccagc
     2101 aacggccaga tcaccaacct gctgcgcggc ttctggggcc tgcgcaaggt gcgcgccgag
     2161 aacgaccgcc accacgccct ggacgccgtg gtggtggcct gcagcaccgt ggccatgcag
     2221 cagaagatca cccgcttcgt gcgctacaag gagatgaacg ccttcgacgg taaaaccatc
     2281 gacaaggaga ccggcgaggt gctgcaccag aagacccact tcccccagcc ctgggagttc
     2341 ttcgcccagg aggtgatgat ccgcgtgttc ggcaagcccg acggcaagcc cgagttcgag
     2401 gaggccgaca cccccgagaa gctgcgcacc ctgctggccg agaagctgag cagccgccct
     2461 gaggccgtgc acgagtacgt gactcctctg ttcgtgagcc gcgcccccaa ccgcaagatg
     2521 agcggtcagg gtcacatgga gaccgtgaag agcgccaagc gcctggacga gggcgtgagc
     2581 gtgctgcgcg tgcccctgac ccagctgaag ctgaaggacc tggagaagat ggtgaaccgc
     2641 gagcgcgagc ccaagctgta cgaggccctg aaggcccgcc tggaggccca caaggacgac
     2701 cccgccaagg ccttcgccga gcccttctac aagtacgaca aggccggcaa ccgcacccag
     2761 caggtgaagg ccgtgcgcgt ggagcaggtg cagaagaccg gcgtgtgggt gcgcaaccac
     2821 aacggcatcg ccgacaacgc caccatggtg cgcgtggacg tgttcgagaa gggcgacaag
     2881 tactacctgg tgcccatcta cagctggcag gtggccaagg gcatcctgcc cgaccgcgcc
     2941 gtggtgcagg gcaaggacga ggaggactgg cagctgatcg acgacagctt caacttcaag
     3001 ttcagcctgc accccaacga cctggtggag gtgatcacca agaaggcccg catgttcggc
     3061 tacttcgcca gctgccaccg cggcaccggc aacatcaaca tccgcatcca cgacctggac
     3121 cacaagatcg gcaagaacgg catcctggag ggcatcggcg tgaagaccgc cctgagcttc
     3181 cagaagtacc agatcgacga gctgggcaag gagatccgcc cctgccgcct gaagaagcgc
     3241 cctcctgtgc gcgcggccgc agatccaaaa aagaagagaa aggtagatcc aaaaaagaag
     3301 agaaaggtag atccaaaaaa gaagagaaag gtagatacgg ccgca
//
