LOCUS       sequence-90727-e        2382 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses N-terminus of D10A SpCas9 nickase domain fused to a GyrA 
            intein, flanked by ITRs for AAV packaging. Combine with C-Terminal 
            Split Cas9 Gyra Intein for full length SpCas9 nickase production.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 2382)
  AUTHORS   Fine EJ, Appleton CM, White DE, Brown MT, Deshmukh H, Kemp ML, Bao G
  TITLE     Trans-spliced Cas9 allows cleavage of HBB and CCR5 genes in human 
            cells using compact expression cassettes.
  JOURNAL   Sci Rep. 2015 Jul 1;5:10777. doi: 10.1038/srep10777.
  PUBMED    26126518
REFERENCE   2  (bases 1 to 2382)
  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: "2015-07-1"; volume: "5:10777. doi"; pages: " 
            10.1038/srep10777"
COMMENT     Sequence Label: N-Terminal Split Cas9 D10A Nickase with GyrA i...
FEATURES             Location/Qualifiers
     source          1..2382
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             1..27
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             31..51
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             61..1776
                     /codon_start=1
                     /product="N-terminal portion of Streptococcus pyogenes Cas9
                     (Zetsche et al., 2015)"
                     /label=Cas9(N)
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIE"
ORIGIN
        1 tacccatacg atgttccaga ttacgcttcg ccgaagaaaa agcgcaaggt cgaagcgtcc
       61 gacaagaagt acagcatcgg cctggccatc ggcaccaact ctgtgggctg ggccgtgatc
      121 accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac
      181 agcatcaaga agaacctgat cggagccctg ctgttcgaca gcggcgaaac agccgaggcc
      241 acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg gatctgctat
      301 ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccacagactg
      361 gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat cttcggcaac
      421 atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gagaaagaaa
      481 ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct ggcccacatg
      541 atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg
      601 gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga aaaccccatc
      661 aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa gagcagacgg
      721 ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt cggcaacctg
      781 attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggat
      841 gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag
      901 atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga cgccatcctg
      961 ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag cgcctctatg
     1021 atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag
     1081 cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg ctacgccggc
     1141 tacattgacg gcggagccag ccaggaagag ttctacaagt tcatcaagcc catcctggaa
     1201 aagatggacg gcaccgagga actgctcgtg aagctgaaca gagaggacct gctgcggaag
     1261 cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga gctgcacgcc
     1321 attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga aaagatcgag
     1381 aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg aaacagcaga
     1441 ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc cctggaactt cgaggaagtg
     1501 gtggacaagg gcgcttccgc ccagagcttc atcgagcgga tgaccaactt cgataagaac
     1561 ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtat
     1621 aacgagctga ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc cttcctgagc
     1681 ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg
     1741 aagcagctga aagaggacta cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc
     1801 ggcgtggaag atcggttcaa cgcctccctg ggcacatacc acgatctgct gaaaattatc
     1861 aaggacaagg acttcctgga caatgaggaa aacgaggaca ttctggaaga tatcgtgctg
     1921 accctgacac tgtttgagga cagagagatg atcgaggaac ggctgaaaac ctatgcccac
     1981 ctgttcgacg acaaagtgat gaagcagctg aagcggcgga gatactgtat caccggtgat
     2041 gctctggtgg cgctgcccga aggcgaatca gtgcgcatcg ctgacatcgt cccgggcgcc
     2101 cgccctaata gcgataacgc catcgacctg aaggtgcttg accgccacgg caatcctgtt
     2161 ctggccgata gactcttcca cagtggtgaa catcctgtgt ataccgtccg caccgtagaa
     2221 ggcctgagag tgacaggcac agccaaccac ccccttttgt gtctggtgga tgttgccggc
     2281 gtccccaccc tgctgtggaa actgattgat gagataaaac ccggagacta tgcagtgatc
     2341 caaaggtccg catttagtgt ggattgcgct ggcttcgctc gc
//
