LOCUS       sequence-410325-        4176 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Cloning A.thaliana codon optimized SpCas9 CDS as GreenGate module B 
            .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4176)
  AUTHORS   Dona M, Bradamante G, Bogojevic Z, Gutzat R, Streubel S, Mosiolek M,
            Dolan L, Mittelsten Scheid O
  TITLE     A versatile CRISPR-based system for lineage tracing in living 
            plants.
  JOURNAL   Plant J. 2023 Jul 5. doi: 10.1111/tpj.16378.
  PUBMED    37403571
REFERENCE   2  (bases 1 to 4176)
  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: "Plant J. 
            2023 Jul 5. doi: 10.1111/tpj.16378."
COMMENT     Sequence Label: pGGB-SpCas9
FEATURES             Location/Qualifiers
     source          1..4176
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             18..4121
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             18..4121
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             4134..4154
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 ggtggtggtc tccaacaatg gataagaagt actctatcgg actcgatatc ggaactaact
       61 ctgtgggatg ggctgtgatc accgatgagt acaaggtgcc atctaagaag ttcaaggttc
      121 tcggaaacac cgataggcac tctatcaaga aaaaccttat cggtgctctc ctcttcgatt
      181 ctggtgaaac tgctgaggct accagactca agagaaccgc tagaagaagg tacaccagaa
      241 gaaagaacag gatctgctac ctccaagaga tcttctctaa cgagatggct aaagtggatg
      301 attcattctt ccacaggctc gaagagtcat tcctcgtgga agaagataag aagcacgaga
      361 ggcaccctat cttcggaaac atcgttgatg aggtggcata ccacgagaag taccctacta
      421 tctaccacct cagaaagaag ctcgttgatt ctactgataa ggctgatctc aggctcatct
      481 acctcgctct cgctcacatg atcaagttca gaggacactt cctcatcgag ggtgatctca
      541 accctgataa ctctgatgtg gataagttgt tcatccagct cgtgcagacc tacaaccagc
      601 ttttcgaaga gaaccctatc aacgcttcag gtgtggatgc taaggctatc ctctctgcta
      661 ggctctctaa gtcaagaagg cttgagaacc tcattgctca gctccctggt gagaagaaga
      721 acggactttt cggaaacttg atcgctctct ctctcggact cacccctaac ttcaagtcta
      781 acttcgatct cgctgaggat gcaaagctcc agctctcaaa ggatacctac gatgatgatc
      841 tcgataacct cctcgctcag atcggagatc agtacgctga tttgttcctc gctgctaaga
      901 acctctctga tgctatcctc ctcagtgata tcctcagagt gaacaccgag atcaccaagg
      961 ctccactctc agcttctatg atcaagagat acgatgagca ccaccaggat ctcacacttc
     1021 tcaaggctct tgttagacag cagctcccag agaagtacaa agagattttc ttcgatcagt
     1081 ctaagaacgg atacgctggt tacatcgatg gtggtgcatc tcaagaagag ttctacaagt
     1141 tcatcaagcc tatcctcgag aagatggatg gaaccgagga actcctcgtg aagctcaata
     1201 gagaggatct tctcagaaag cagaggacct tcgataacgg atctatccct catcagatcc
     1261 acctcggaga gttgcacgct atccttagaa ggcaagagga tttctaccca ttcctcaagg
     1321 ataacaggga aaagattgag aagattctca ccttcagaat cccttactac gtgggacctc
     1381 tcgctagagg aaactcaaga ttcgcttgga tgaccagaaa gtctgaggaa accatcaccc
     1441 cttggaactt cgaagaggtg gtggataagg gtgctagtgc tcagtctttc atcgagagga
     1501 tgaccaactt cgataagaac cttccaaacg agaaggtgct ccctaagcac tctttgctct
     1561 acgagtactt caccgtgtac aacgagttga ccaaggttaa gtacgtgacc gagggaatga
     1621 ggaagcctgc ttttttgtca ggtgagcaaa agaaggctat cgttgatctc ttgttcaaga
     1681 ccaacagaaa ggtgaccgtg aagcagctca aagaggatta cttcaagaaa atcgagtgct
     1741 tcgattcagt tgagatttct ggtgttgagg ataggttcaa cgcatctctc ggaacctacc
     1801 acgatctcct caagatcatt aaggataagg atttcttgga taacgaggaa aacgaggata
     1861 tcttggagga tatcgttctt accctcaccc tctttgaaga tagagagatg attgaagaaa
     1921 ggctcaagac ctacgctcat ctcttcgatg ataaggtgat gaagcagttg aagagaagaa
     1981 gatacactgg ttggggaagg ctctcaagaa agctcattaa cggaatcagg gataagcagt
     2041 ctggaaagac aatccttgat ttcctcaagt ctgatggatt cgctaacaga aacttcatgc
     2101 agctcatcca cgatgattct ctcaccttta aagaggatat ccagaaggct caggtttcag
     2161 gacagggtga tagtctccat gagcatatcg ctaacctcgc tggatctcct gcaatcaaga
     2221 agggaatcct ccagactgtg aaggttgtgg atgagttggt gaaggtgatg ggaaggcata
     2281 agcctgagaa catcgtgatc gaaatggcta gagagaacca gaccactcag aagggacaga
     2341 agaactctag ggaaaggatg aagaggatcg aggaaggtat caaagagctt ggatctcaga
     2401 tcctcaaaga gcaccctgtt gagaacactc agctccagaa tgagaagctc tacctctact
     2461 acctccagaa cggaagggat atgtatgtgg atcaagagtt ggatatcaac aggctctctg
     2521 attacgatgt tgatcatatc gtgccacagt cattcttgaa ggatgattct atcgataaca
     2581 aggtgctcac caggtctgat aagaacaggg gtaagagtga taacgtgcca agtgaagagg
     2641 ttgtgaagaa aatgaagaac tattggaggc agctcctcaa cgctaagctc atcactcaga
     2701 gaaagttcga taacttgact aaggctgaga ggggaggact ctctgaattg gataaggcag
     2761 gattcatcaa gaggcagctt gtggaaacca ggcagatcac taagcacgtt gcacagatcc
     2821 tcgattctag gatgaacacc aagtacgatg agaacgataa gttgatcagg gaagtgaagg
     2881 ttatcaccct caagtcaaag ctcgtgtctg atttcagaaa ggatttccaa ttctacaagg
     2941 tgagggaaat caacaactac caccacgctc acgatgctta ccttaacgct gttgttggaa
     3001 ccgctctcat caagaagtat cctaagctcg agtcagagtt cgtgtacggt gattacaagg
     3061 tgtacgatgt gaggaagatg atcgctaagt ctgagcaaga gatcggaaag gctaccgcta
     3121 agtatttctt ctactctaac atcatgaatt tcttcaagac cgagattacc ctcgctaacg
     3181 gtgagatcag aaagaggcca ctcatcgaga caaacggtga aacaggtgag atcgtgtggg
     3241 ataagggaag ggatttcgct accgttagaa aggtgctctc tatgccacag gtgaacatcg
     3301 ttaagaaaac cgaggtgcag accggtggat tctctaaaga gtctatcctc cctaagagga
     3361 actctgataa gctcattgct aggaagaagg attgggaccc taagaaatac ggtggtttcg
     3421 attctcctac cgtggcttac tctgttctcg ttgtggctaa ggttgagaag ggaaagagta
     3481 agaagctcaa gtctgttaag gaacttctcg gaatcactat catggaaagg tcatctttcg
     3541 agaagaaccc aatcgatttc ctcgaggcta agggatacaa agaggttaag aaggatctca
     3601 tcatcaagct cccaaagtac tcactcttcg aactcgagaa cggtagaaag aggatgctcg
     3661 cttctgctgg tgagcttcaa aagggaaacg agcttgctct cccatctaag tacgttaact
     3721 ttctttacct cgcttctcac tacgagaagt tgaagggatc tccagaagat aacgagcaga
     3781 agcaactttt cgttgagcag cacaagcact acttggatga gatcatcgag cagatctctg
     3841 agttctctaa aagggtgatc ctcgctgatg caaacctcga taaggtgttg tctgcttaca
     3901 acaagcacag agataagcct atcagggaac aggcagagaa catcatccat ctcttcaccc
     3961 ttaccaacct cggtgctcct gctgctttca agtacttcga tacaaccatc gataggaaga
     4021 gatacacctc taccaaagaa gtgctcgatg ctaccctcat ccatcagtct atcactggac
     4081 tctacgagac taggatcgat ctctcacagc tcggtggtga ttcaagggct gatcctaaga
     4141 agaagaggaa ggtttgaggg gctcgagacc ggtggt
//
