LOCUS       sequence-260236-        4149 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression fusion protein ALKBH5-dCas9 in Mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4149)
  AUTHORS   Liu XM, Zhou J, Mao Y, Ji Q, Qian SB
  TITLE     Programmable RNA N(6)-methyladenosine editing by CRISPR-Cas9 
            conjugates.
  JOURNAL   Nat Chem Biol. 2019 Sep;15(9):865-871. doi: 
            10.1038/s41589-019-0327-1. Epub 2019 Aug 5.
  PUBMED    31383972
REFERENCE   2  (bases 1 to 4149)
  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/s41589-019-0327-1"; journalName: "Nat Chem Biol"; date: 
            "2019-09"; volume: "15"; issue: "9"; pages: "865-871"
COMMENT     Sequence Label: ALKBH5-dCas9
FEATURES             Location/Qualifiers
     source          1..4149
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             join(49..3447,3448..3450,3451..4047,4048,4049..4051,
                     4052..4053,4054..4056,4057..4149)
                     /codon_start=1
                     /product="Streptococcus pyogenes Cas9 endonuclease with the
                     D1135V/R1335Q/T1337R mutations"
                     /label=Cas9 VQR
                     /note="has an altered recognition specificity for the 
                     protospacer adjacent motif (PAM) (Kleinstiver et al., 
                     2015)"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             join(49..72,73..75,76..2562,2563..2565,2566..4149)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
ORIGIN
        1 agcggcagcg agactcccgg gacctcagag tccgccacac ccgaaagtga taaaaagtat
       61 tctattggtt tagccatcgg cactaattcc gttggatggg ctgtcataac cgatgaatac
      121 aaagtacctt caaagaaatt taaggtgttg gggaacacag accgtcattc gattaaaaag
      181 aatcttatcg gtgccctcct attcgatagt ggcgaaacgg cagaggcgac tcgcctgaaa
      241 cgaaccgctc ggagaaggta tacacgtcgc aagaaccgaa tatgttactt acaagaaatt
      301 tttagcaatg agatggccaa agttgacgat tctttctttc accgtttgga agagtccttc
      361 cttgtcgaag aggacaagaa acatgaacgg caccccatct ttggaaacat agtagatgag
      421 gtggcatatc atgaaaagta cccaacgatt tatcacctca gaaaaaagct agttgactca
      481 actgataaag cggacctgag gttaatctac ttggctcttg cccatatgat aaagttccgt
      541 gggcactttc tcattgaggg tgatctaaat ccggacaact cggatgtcga caaactgttc
      601 atccagttag tacaaaccta taatcagttg tttgaagaga accctataaa tgcaagtggc
      661 gtggatgcga aggctattct tagcgcccgc ctctctaaat cccgacggct agaaaacctg
      721 atcgcacaat tacccggaga gaagaaaaat gggttgttcg gtaaccttat agcgctctca
      781 ctaggcctga caccaaattt taagtcgaac ttcgacttag ctgaagatgc caaattgcag
      841 cttagtaagg acacgtacga tgacgatctc gacaatctac tggcacaaat tggagatcag
      901 tatgcggact tatttttggc tgccaaaaac cttagcgatg caatcctcct atctgacata
      961 ctgagagtta atactgagat taccaaggcg ccgttatccg cttcaatgat caaaaggtac
     1021 gatgaacatc accaagactt gacacttctc aaggccctag tccgtcagca actgcctgag
     1081 aaatataagg aaatattctt tgatcagtcg aaaaacgggt acgcaggtta tattgacggc
     1141 ggagcgagtc aagaggaatt ctacaagttt atcaaaccca tattagagaa gatggatggg
     1201 acggaagagt tgcttgtaaa actcaatcgc gaagatctac tgcgaaagca gcggactttc
     1261 gacaacggta gcattccaca tcaaatccac ttaggcgaat tgcatgctat acttagaagg
     1321 caggaggatt tttatccgtt cctcaaagac aatcgtgaaa agattgagaa aatcctaacc
     1381 tttcgcatac cttactatgt gggacccctg gcccgaggga actctcggtt cgcatggatg
     1441 acaagaaagt ccgaagaaac gattactcca tggaattttg aggaagttgt cgataaaggt
     1501 gcgtcagctc aatcgttcat cgagaggatg accaactttg acaagaattt accgaacgaa
     1561 aaagtattgc ctaagcacag tttactttac gagtatttca cagtgtacaa tgaactcacg
     1621 aaagttaagt atgtcactga gggcatgcgt aaacccgcct ttctaagcgg agaacagaag
     1681 aaagcaatag tagatctgtt attcaagacc aaccgcaaag tgacagttaa gcaattgaaa
     1741 gaggactact ttaagaaaat tgaatgcttc gattctgtcg agatctccgg ggtagaagat
     1801 cgatttaatg cgtcacttgg tacgtatcat gacctcctaa agataattaa agataaggac
     1861 ttcctggata acgaagagaa tgaagatatc ttagaagata tagtgttgac tcttaccctc
     1921 tttgaagatc gggaaatgat tgaggaaaga ctaaaaacat acgctcacct gttcgacgat
     1981 aaggttatga aacagttaaa gaggcgtcgc tatacgggct ggggacgatt gtcgcggaaa
     2041 cttatcaacg ggataagaga caagcaaagt ggtaaaacta ttctcgattt tctaaagagc
     2101 gacggcttcg ccaataggaa ctttatgcag ctgatccatg atgactcttt aaccttcaaa
     2161 gaggatatac aaaaggcaca ggtttccgga caaggggact cattgcacga acatattgcg
     2221 aatcttgctg gttcgccagc catcaaaaag ggcatactcc agacagtcaa agtagtggat
     2281 gagctagtta aggtcatggg acgtcacaaa ccggaaaaca ttgtaatcga gatggcacgc
     2341 gaaaatcaaa cgactcagaa ggggcaaaaa aacagtcgag agcggatgaa gagaatagaa
     2401 gagggtatta aagaactggg cagccagatc ttaaaggagc atcctgtgga aaatacccaa
     2461 ttgcagaacg agaaacttta cctctattac ctacaaaatg gaagggacat gtatgttgat
     2521 caggaactgg acataaaccg tttatctgat tacgacgtcg atgccattgt accccaatcc
     2581 tttttgaagg acgattcaat cgacaataaa gtgcttacac gctcggataa gaaccgaggg
     2641 aaaagtgaca atgttccaag cgaggaagtc gtaaagaaaa tgaagaacta ttggcggcag
     2701 ctcctaaatg cgaaactgat aacgcaaaga aagttcgata acttaactaa agctgagagg
     2761 ggtggcttgt ctgaacttga caaggccgga tttattaaac gtcagctcgt ggaaacccgc
     2821 caaatcacaa agcatgttgc acagatacta gattcccgaa tgaatacgaa atacgacgag
     2881 aacgataagc tgattcggga agtcaaagta atcactttaa agtcaaaatt ggtgtcggac
     2941 ttcagaaagg attttcaatt ctataaagtt agggagataa ataactacca ccatgcgcac
     3001 gacgcttatc ttaatgccgt cgtagggacc gcactcatta agaaataccc gaagctagaa
     3061 agtgagtttg tgtatggtga ttacaaagtt tatgacgtcc gtaagatgat cgcgaaaagc
     3121 gaacaggaga taggcaaggc tacagccaaa tacttctttt attctaacat tatgaatttc
     3181 tttaagacgg aaatcactct ggcaaacgga gagatacgca aacgaccttt aattgaaacc
     3241 aatggggaga caggtgaaat cgtatgggat aagggccggg acttcgcgac ggtgagaaaa
     3301 gttttgtcca tgccccaagt caacatagta aagaaaactg aggtgcagac cggagggttt
     3361 tcaaaggaat cgattcttcc aaaaaggaat agtgataagc tcatcgctcg taaaaaggac
     3421 tgggacccga aaaagtacgg tggcttcgat agccctacag ttgcctattc tgtcctagta
     3481 gtggcaaaag ttgagaaggg aaaatccaag aaactgaagt cagtcaaaga attattgggg
     3541 ataacgatta tggagcgctc gtcttttgaa aagaacccca tcgacttcct tgaggcgaaa
     3601 ggttacaagg aagtaaaaaa ggatctcata attaaactac caaagtatag tctgtttgag
     3661 ttagaaaatg gccgaaaacg gatgttggct agcgccggag agcttcaaaa ggggaacgaa
     3721 ctcgcactac cgtctaaata cgtgaatttc ctgtatttag cgtcccatta cgagaagttg
     3781 aaaggttcac ctgaagataa cgaacagaag caactttttg ttgagcagca caaacattat
     3841 ctcgacgaaa tcatagagca aatttcggaa ttcagtaaga gagtcatcct agctgatgcc
     3901 aatctggaca aagtattaag cgcatacaac aagcacaggg ataaacccat acgtgagcag
     3961 gcggaaaata ttatccattt gtttactctt accaacctcg gcgctccagc cgcattcaag
     4021 tattttgaca caacgataga tcgcaaacga tacacttcta ccaaggaggt gctagacgcg
     4081 acactgattc accaatccat cacgggatta tatgaaactc ggatagattt gtcacagctt
     4141 gggggtgac
//
