LOCUS       sequence-218168-        5403 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 5403)
  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 5403)
  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-5
FEATURES             Location/Qualifiers
     source          1..5403
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             4..24
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(37..60,61..63,64..4137)
                     /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             4050..4058
                     /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             4144..4164
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5353..5400
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
ORIGIN
        1 atgccaaaaa agaagagaaa ggtttcaggc ggctccgaca agaagtactc gatcggcctc
       61 gccattggga ctaactctgt tggctgggcc gtgatcaccg acgagtacaa ggtgccctca
      121 aagaagttca aggtcctggg caacaccgat cggcattcca tcaagaagaa tctcattggc
      181 gctctcctgt tcgacagcgg cgagacggct gaggctacgc ggctcaagcg caccgcccgc
      241 aggcggtaca cgcgcaggaa gaatcgcatc tgctacctgc aggagatttt ctccaacgag
      301 atggcgaagg ttgacgattc tttcttccac aggctggagg agtcattcct cgtggaggag
      361 gataagaagc acgagcggca tccaatcttc ggcaacattg tcgacgaggt tgcctaccac
      421 gagaagtacc ctacgatcta ccatctgcgg aagaagctcg tggactccac agataaggcg
      481 gacctccgcc tgatctacct cgctctggcc cacatgatta agttcagggg ccatttcctg
      541 atcgaggggg atctcaaccc ggacaatagc gatgttgaca agctgttcat ccagctcgtg
      601 cagacgtaca accagctctt cgaggagaac cccattaatg cgtcaggcgt cgacgcgaag
      661 gctatcctgt ccgctaggct ctcgaagtct cggcgcctcg agaacctgat cgcccagctg
      721 ccgggcgaga agaagaacgg cctgttcggg aatctcattg cgctcagcct ggggctcacg
      781 cccaacttca agtcgaattt cgatctcgct gaggacgcca agctgcagct ctccaaggac
      841 acatacgacg atgacctgga taacctcctg gcccagatcg gcgatcagta cgcggacctg
      901 ttcctcgctg ccaagaatct gtcggacgcc atcctcctgt ctgatattct cagggtgaac
      961 accgagatta cgaaggctcc gctctcagcc tccatgatca agcgctacga cgagcaccat
     1021 caggatctga ccctcctgaa ggcgctggtc aggcagcagc tccccgagaa gtacaaggag
     1081 atcttcttcg atcagtcgaa gaacggctac gctgggtaca ttgacggcgg ggcctctcag
     1141 gaggagttct acaagttcat caagccgatt ctggagaaga tggacggcac ggaggagctg
     1201 ctggtgaagc tcaatcgcga ggacctcctg aggaagcagc ggacattcga taacggcagc
     1261 atcccacacc agattcatct cggggagctg cacgctatcc tgaggaggca ggaggacttc
     1321 taccctttcc tcaaggataa ccgcgagaag atcgagaaga ttctgacttt caggatcccg
     1381 tactacgtcg gcccactcgc taggggcaac tcccgcttcg cttggatgac ccgcaagtca
     1441 gaggagacga tcacgccgtg gaacttcgag gaggtggtcg acaagggcgc tagcgctcag
     1501 tcgttcatcg agaggatgac gaatttcgac aagaacctgc caaatgagaa ggtgctccct
     1561 aagcactcgc tcctgtacga gtacttcaca gtctacaacg agctgactaa ggtgaagtat
     1621 gtgaccgagg gcatgaggaa gccggctttc ctgtctgggg agcagaagaa ggccatcgtg
     1681 gacctcctgt tcaagaccaa ccggaaggtc acggttaagc agctcaagga ggactacttc
     1741 aagaagattg agtgcttcga ttcggtcgag atctctggcg ttgaggaccg cttcaacgcc
     1801 tccctgggga cctaccacga tctcctgaag atcattaagg ataaggactt cctggacaac
     1861 gaggagaatg aggatatcct cgaggacatt gtgctgacac tcactctgtt cgaggaccgg
     1921 gagatgatcg aggagcgcct gaagacttac gcccatctct tcgatgacaa ggtcatgaag
     1981 cagctcaaga ggaggaggta caccggctgg gggaggctga gcaggaagct catcaacggc
     2041 attcgggaca agcagtccgg gaagacgatc ctcgacttcc tgaagagcga tggcttcgcg
     2101 aaccgcaatt tcatgcagct gattcacgat gacagcctca cattcaagga ggatatccag
     2161 aaggctcagg tgagcggcca gggggactcg ctgcacgagc atatcgcgaa cctcgctggc
     2221 tcgccagcta tcaagaaggg gattctgcag accgtgaagg ttgtggacga gctggtgaag
     2281 gtcatgggca ggcacaagcc tgagaacatc gtcattgaga tggcccggga gaatcagacc
     2341 acgcagaagg gccagaagaa ctcacgcgag aggatgaaga ggatcgagga gggcattaag
     2401 gagctggggt cccagatcct caaggagcac ccggtggaga acacgcagct gcagaatgag
     2461 aagctctacc tgtactacct ccagaatggc cgcgatatgt atgtggacca ggagctggat
     2521 attaacaggc tcagcgatta cgacgtcgat catatcgttc cacagtcatt cctgaaggat
     2581 gactccattg acaacaaggt cctcaccagg tcggacaaga accggggcaa gtctgataat
     2641 gttccttcag aggaggtcgt taagaagatg aagaactact ggcgccagct cctgaatgcc
     2701 aagctgatca cgcagcggaa gttcgataac ctcacaaagg ctgagagggg cgggctctct
     2761 gagctggaca aggcgggctt catcaagagg cagctggtcg agacacggca gatcactaag
     2821 cacgttgcgc agattctcga ctcacggatg aacactaagt acgatgagaa tgacaagctg
     2881 atccgcgagg tgaaggtcat caccctgaag tcaaagctcg tctccgactt caggaaggat
     2941 ttccagttct acaaggttcg ggagatcaac aattaccacc atgcccatga cgcgtacctg
     3001 aacgcggtgg tcggcacagc tctgatcaag aagtacccaa agctcgagag cgagttcgtg
     3061 tacggggact acaaggttta cgatgtgagg aagatgatcg ccaagtcgga gcaggagatt
     3121 ggcaaggcta ccgccaagta cttcttctac tctaacatta tgaatttctt caagacagag
     3181 atcactctgg ccaatggcga gatccggaag cgccccctca tcgagacgaa cggcgagacg
     3241 ggggagatcg tgtgggacaa gggcagggat ttcgcgaccg tcaggaaggt tctctccatg
     3301 ccacaagtga atatcgtcaa gaagacagag gtccagactg gcgggttctc taaggagtca
     3361 attctgccta agcggaacag cgacaagctc atcgcccgca agaaggactg ggatccgaag
     3421 aagtacggcg ggttcgacag ccccactgtg gcctactcgg tcctggttgt ggcgaaggtt
     3481 gagaagggca agtccaagaa gctcaagagc gtgaaggagc tgctggggat cacgattatg
     3541 gagcgctcca gcttcgagaa gaacccgatc gatttcctgg aggcgaaggg ctacaaggag
     3601 gtgaagaagg acctgatcat taagctcccc aagtactcac tcttcgagct ggagaacggc
     3661 aggaagcgga tgctggcttc cgctggcgag ctgcagaagg ggaacgagct ggctctgccg
     3721 tccaagtatg tgaacttcct ctacctggcc tcccactacg agaagctcaa gggcagcccc
     3781 gaggacaacg agcagaagca gctgttcgtc gagcagcaca agcattacct cgacgagatc
     3841 attgagcaga tttccgagtt ctccaagcgc gtgatcctgg ccgacgcgaa tctggataag
     3901 gtcctctccg cgtacaacaa gcaccgcgac aagccaatca gggagcaggc tgagaatatc
     3961 attcatctct tcaccctgac gaacctcggc gcccctgctg ctttcaagta cttcgacaca
     4021 actatcgatc gcaagaggta cacaagcact aaggaggtcc tggacgcgac cctcatccac
     4081 cagtcgatta ccggcctcta cgagacgcgc atcgacctgt ctcagctcgg gggcgacgaa
     4141 ttcccaaaga agaagcgcaa ggtctccggc ggatcttctg gaggatctag cggctccgag
     4201 acaccaggaa catccgaatc cgctacacca gagtcttctg gaggatctag cggaggatct
     4261 tccgaggtgg agttctctca cgagtattgg atgaggcacg ctcttacact tgctaagaga
     4321 gcttgggacg aaagagaagt gccagttggc gccgttcttg tgcataataa tagggtgatc
     4381 ggcgagggtt ggaatagacc aattggaagg catgatccaa cagctcacgc agagattatg
     4441 gctctcagac aaggcggcct cgttatgcag aactacaggc tcattgacgc tacactctac
     4501 gtgacactcg aaccttgcgt tatgtgcgcc ggagctatga ttcattctag gattggcagg
     4561 gtcgtgtttg gagctaggga cgctaaaaca ggagccgccg gatctcttat ggacgtgttg
     4621 catcatccag gcatgaacca tagggtggag attacagagg gcattcttgc agacgagtgc
     4681 gctgctcttc tttccgattt cttcaggatg agaaggcagg agattaaggc ccagaagaag
     4741 gctcagtctt ctacagatag cggaggatct tccggaggat ctagcggctc cgagacacca
     4801 ggaacatccg aaagcgctac accagaatct agcggaggct cttccggagg atcttctgaa
     4861 gtggagttct cccacgagta ttggatgagg cacgctctta cacttgctaa aagggctagg
     4921 gacgaaaggg aagttccagt tggagctgtt ctcgtgctca ataacagggt gattggcgag
     4981 ggttggaata gagccattgg actccatgat ccaacagctc acgcagagat tatggctctt
     5041 agacaaggcg gcctcgttat gcagaattac agactcatcg acgccacact ctacgttacc
     5101 ttcgaacctt gcgttatgtg cgccggagct atgatccatt ctaggattgg cagggtcgtg
     5161 ttcggcgtta gaaacgctaa gacaggagct gcaggctctc ttatggacgt tcttcattac
     5221 ccaggcatga atcatagagt ggagatcaca gaaggcattc ttgcagacga gtgcgcagct
     5281 ctcctttgct atttcttcag gatgccgagg caagttttca acgctcagaa gaaggcccag
     5341 tcttctacag ataagagacc agcagctaca aagaaggccg gacaagctaa gaagaagaag
     5401 tag
//
