LOCUS       sequence-218172-        5400 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Targeted A to G in wheat plants or monocotyledons protoplasts .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5400)
  AUTHORS   Li C, Zong Y, Wang Y, Jin S, Zhang D, Song Q, Zhang R, Gao C
  TITLE     Expanded base editing in rice and wheat using a Cas9-adenosine 
            deaminase fusion.
  JOURNAL   Genome Biol. 2018 May 29;19(1):59. doi: 10.1186/s13059-018-1443-z.
  PUBMED    29807545
REFERENCE   2  (bases 1 to 5400)
  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: "Genome 
            Biol."; date: "2018-05-29"; volume: "19(1):59. doi"; pages: " 
            10.1186/s13059-018-1443-z"
COMMENT     Sequence Label: PABE-7
FEATURES             Location/Qualifiers
     source          1..5400
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             join(1198..1221,1222..1224,1225..5298)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     RBS             5211..5219
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             5311..5331
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5344..5364
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5377..5397
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 atgtccgagg tggagttctc tcacgagtat tggatgaggc acgctcttac acttgctaag
       61 agagcttggg acgaaagaga agtgccagtt ggcgccgttc ttgtgcataa taatagggtg
      121 atcggcgagg gttggaatag accaattgga aggcatgatc caacagctca cgcagagatt
      181 atggctctca gacaaggcgg cctcgttatg cagaactaca ggctcattga cgctacactc
      241 tacgtgacac tcgaaccttg cgttatgtgc gccggagcta tgattcattc taggattggc
      301 agggtcgtgt ttggagctag ggacgctaaa acaggagccg ccggatctct tatggacgtg
      361 ttgcatcatc caggcatgaa ccatagggtg gagattacag agggcattct tgcagacgag
      421 tgcgctgctc ttctttccga tttcttcagg atgagaaggc aggagattaa ggcccagaag
      481 aaggctcagt cttctacaga tagcggagga tcttccggag gatctagcgg ctccgagaca
      541 ccaggaacat ccgaaagcgc tacaccagaa tctagcggag gctcttccgg aggatcttct
      601 gaagtggagt tctcccacga gtattggatg aggcacgctc ttacacttgc taaaagggct
      661 agggacgaaa gggaagttcc agttggagct gttctcgtgc tcaataacag ggtgattggc
      721 gagggttgga atagagccat tggactccat gatccaacag ctcacgcaga gattatggct
      781 cttagacaag gcggcctcgt tatgcagaat tacagactca tcgacgccac actctacgtt
      841 accttcgaac cttgcgttat gtgcgccgga gctatgatcc attctaggat tggcagggtc
      901 gtgttcggcg ttagaaacgc taagacagga gctgcaggct ctcttatgga cgttcttcat
      961 tacccaggca tgaatcatag agtggagatc acagaaggca ttcttgcaga cgagtgcgca
     1021 gctctccttt gctatttctt caggatgccg aggcaagttt tcaacgctca gaagaaggcc
     1081 cagtcttcta cagattccgg cggatcttct ggaggatcta gcggctccga gacaccagga
     1141 acatccgaat ccgctacacc agagtcttct ggaggatcta gcggaggatc tcttaaggac
     1201 aagaagtact cgatcggcct cgccattggg actaactctg ttggctgggc cgtgatcacc
     1261 gacgagtaca aggtgccctc aaagaagttc aaggtcctgg gcaacaccga tcggcattcc
     1321 atcaagaaga atctcattgg cgctctcctg ttcgacagcg gcgagacggc tgaggctacg
     1381 cggctcaagc gcaccgcccg caggcggtac acgcgcagga agaatcgcat ctgctacctg
     1441 caggagattt tctccaacga gatggcgaag gttgacgatt ctttcttcca caggctggag
     1501 gagtcattcc tcgtggagga ggataagaag cacgagcggc atccaatctt cggcaacatt
     1561 gtcgacgagg ttgcctacca cgagaagtac cctacgatct accatctgcg gaagaagctc
     1621 gtggactcca cagataaggc ggacctccgc ctgatctacc tcgctctggc ccacatgatt
     1681 aagttcaggg gccatttcct gatcgagggg gatctcaacc cggacaatag cgatgttgac
     1741 aagctgttca tccagctcgt gcagacgtac aaccagctct tcgaggagaa ccccattaat
     1801 gcgtcaggcg tcgacgcgaa ggctatcctg tccgctaggc tctcgaagtc tcggcgcctc
     1861 gagaacctga tcgcccagct gccgggcgag aagaagaacg gcctgttcgg gaatctcatt
     1921 gcgctcagcc tggggctcac gcccaacttc aagtcgaatt tcgatctcgc tgaggacgcc
     1981 aagctgcagc tctccaagga cacatacgac gatgacctgg ataacctcct ggcccagatc
     2041 ggcgatcagt acgcggacct gttcctcgct gccaagaatc tgtcggacgc catcctcctg
     2101 tctgatattc tcagggtgaa caccgagatt acgaaggctc cgctctcagc ctccatgatc
     2161 aagcgctacg acgagcacca tcaggatctg accctcctga aggcgctggt caggcagcag
     2221 ctccccgaga agtacaagga gatcttcttc gatcagtcga agaacggcta cgctgggtac
     2281 attgacggcg gggcctctca ggaggagttc tacaagttca tcaagccgat tctggagaag
     2341 atggacggca cggaggagct gctggtgaag ctcaatcgcg aggacctcct gaggaagcag
     2401 cggacattcg ataacggcag catcccacac cagattcatc tcggggagct gcacgctatc
     2461 ctgaggaggc aggaggactt ctaccctttc ctcaaggata accgcgagaa gatcgagaag
     2521 attctgactt tcaggatccc gtactacgtc ggcccactcg ctaggggcaa ctcccgcttc
     2581 gcttggatga cccgcaagtc agaggagacg atcacgccgt ggaacttcga ggaggtggtc
     2641 gacaagggcg ctagcgctca gtcgttcatc gagaggatga cgaatttcga caagaacctg
     2701 ccaaatgaga aggtgctccc taagcactcg ctcctgtacg agtacttcac agtctacaac
     2761 gagctgacta aggtgaagta tgtgaccgag ggcatgagga agccggcttt cctgtctggg
     2821 gagcagaaga aggccatcgt ggacctcctg ttcaagacca accggaaggt cacggttaag
     2881 cagctcaagg aggactactt caagaagatt gagtgcttcg attcggtcga gatctctggc
     2941 gttgaggacc gcttcaacgc ctccctgggg acctaccacg atctcctgaa gatcattaag
     3001 gataaggact tcctggacaa cgaggagaat gaggatatcc tcgaggacat tgtgctgaca
     3061 ctcactctgt tcgaggaccg ggagatgatc gaggagcgcc tgaagactta cgcccatctc
     3121 ttcgatgaca aggtcatgaa gcagctcaag aggaggaggt acaccggctg ggggaggctg
     3181 agcaggaagc tcatcaacgg cattcgggac aagcagtccg ggaagacgat cctcgacttc
     3241 ctgaagagcg atggcttcgc gaaccgcaat ttcatgcagc tgattcacga tgacagcctc
     3301 acattcaagg aggatatcca gaaggctcag gtgagcggcc agggggactc gctgcacgag
     3361 catatcgcga acctcgctgg ctcgccagct atcaagaagg ggattctgca gaccgtgaag
     3421 gttgtggacg agctggtgaa ggtcatgggc aggcacaagc ctgagaacat cgtcattgag
     3481 atggcccggg agaatcagac cacgcagaag ggccagaaga actcacgcga gaggatgaag
     3541 aggatcgagg agggcattaa ggagctgggg tcccagatcc tcaaggagca cccggtggag
     3601 aacacgcagc tgcagaatga gaagctctac ctgtactacc tccagaatgg ccgcgatatg
     3661 tatgtggacc aggagctgga tattaacagg ctcagcgatt acgacgtcga tcatatcgtt
     3721 ccacagtcat tcctgaagga tgactccatt gacaacaagg tcctcaccag gtcggacaag
     3781 aaccggggca agtctgataa tgttccttca gaggaggtcg ttaagaagat gaagaactac
     3841 tggcgccagc tcctgaatgc caagctgatc acgcagcgga agttcgataa cctcacaaag
     3901 gctgagaggg gcgggctctc tgagctggac aaggcgggct tcatcaagag gcagctggtc
     3961 gagacacggc agatcactaa gcacgttgcg cagattctcg actcacggat gaacactaag
     4021 tacgatgaga atgacaagct gatccgcgag gtgaaggtca tcaccctgaa gtcaaagctc
     4081 gtctccgact tcaggaagga tttccagttc tacaaggttc gggagatcaa caattaccac
     4141 catgcccatg acgcgtacct gaacgcggtg gtcggcacag ctctgatcaa gaagtaccca
     4201 aagctcgaga gcgagttcgt gtacggggac tacaaggttt acgatgtgag gaagatgatc
     4261 gccaagtcgg agcaggagat tggcaaggct accgccaagt acttcttcta ctctaacatt
     4321 atgaatttct tcaagacaga gatcactctg gccaatggcg agatccggaa gcgccccctc
     4381 atcgagacga acggcgagac gggggagatc gtgtgggaca agggcaggga tttcgcgacc
     4441 gtcaggaagg ttctctccat gccacaagtg aatatcgtca agaagacaga ggtccagact
     4501 ggcgggttct ctaaggagtc aattctgcct aagcggaaca gcgacaagct catcgcccgc
     4561 aagaaggact gggatccgaa gaagtacggc gggttcgaca gccccactgt ggcctactcg
     4621 gtcctggttg tggcgaaggt tgagaagggc aagtccaaga agctcaagag cgtgaaggag
     4681 ctgctgggga tcacgattat ggagcgctcc agcttcgaga agaacccgat cgatttcctg
     4741 gaggcgaagg gctacaagga ggtgaagaag gacctgatca ttaagctccc caagtactca
     4801 ctcttcgagc tggagaacgg caggaagcgg atgctggctt ccgctggcga gctgcagaag
     4861 gggaacgagc tggctctgcc gtccaagtat gtgaacttcc tctacctggc ctcccactac
     4921 gagaagctca agggcagccc cgaggacaac gagcagaagc agctgttcgt cgagcagcac
     4981 aagcattacc tcgacgagat cattgagcag atttccgagt tctccaagcg cgtgatcctg
     5041 gccgacgcga atctggataa ggtcctctcc gcgtacaaca agcaccgcga caagccaatc
     5101 agggagcagg ctgagaatat cattcatctc ttcaccctga cgaacctcgg cgcccctgct
     5161 gctttcaagt acttcgacac aactatcgat cgcaagaggt acacaagcac taaggaggtc
     5221 ctggacgcga ccctcatcca ccagtcgatt accggcctct acgagacgcg catcgacctg
     5281 tctcagctcg ggggcgactc cggcggcagc ccaaagaaga agcggaaggt gtctggaggt
     5341 tctcctaaga aaaagagaaa agtgtccggc ggctccccga agaagaagcg caaggtgtag
//
