LOCUS       sequence-410330-        5068 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Cloning estradiol-inducible expression cassette containi LexA XVE 
            binding sequence followed by SpCas9 CDS and PEA3AT as GreenGate 
            module C.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5068)
  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 5068)
  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: pGGC-LexACasPea
FEATURES             Location/Qualifiers
     source          1..5068
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        371..417
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             439..4542
                     /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             439..4542
                     /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             4555..4575
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 ggtggtggtc tccggctcct agcaaagtcg acgaattctc ccgggtggat ccaaggcctt
       61 ctagaaaaac gaattccatg ccagcttggg ctgcaggtcg aggctaaaaa actaatcgca
      121 ttatcatccc ctcgacgtac tgtacatata accactggtt ttatatacag cagtactgta
      181 catataacca ctggttttat atacagcagt cgacgtactg tacatataac cactggtttt
      241 atatacagca gtactgtaca tataaccact ggttttatat acagcagtcg aggtaagatt
      301 agatatggat atgtatatgg atatgtatat ggtggtaatg ccatgtaata tgctcgactc
      361 taggatcttc gcaagaccct tcctctatat aaggaagttc atttcatttg gagaggacac
      421 gctgaagcta gtcgctgcat ggataagaag tactctatcg gactcgatat cggaactaac
      481 tctgtgggat gggctgtgat caccgatgag tacaaggtgc catctaagaa gttcaaggtt
      541 ctcggaaaca ccgataggca ctctatcaag aaaaacctta tcggtgctct cctcttcgat
      601 tctggtgaaa ctgctgaggc taccagactc aagagaaccg ctagaagaag gtacaccaga
      661 agaaagaaca ggatctgcta cctccaagag atcttctcta acgagatggc taaagtggat
      721 gattcattct tccacaggct cgaagagtca ttcctcgtgg aagaagataa gaagcacgag
      781 aggcacccta tcttcggaaa catcgttgat gaggtggcat accacgagaa gtaccctact
      841 atctaccacc tcagaaagaa gctcgttgat tctactgata aggctgatct caggctcatc
      901 tacctcgctc tcgctcacat gatcaagttc agaggacact tcctcatcga gggtgatctc
      961 aaccctgata actctgatgt ggataagttg ttcatccagc tcgtgcagac ctacaaccag
     1021 cttttcgaag agaaccctat caacgcttca ggtgtggatg ctaaggctat cctctctgct
     1081 aggctctcta agtcaagaag gcttgagaac ctcattgctc agctccctgg tgagaagaag
     1141 aacggacttt tcggaaactt gatcgctctc tctctcggac tcacccctaa cttcaagtct
     1201 aacttcgatc tcgctgagga tgcaaagctc cagctctcaa aggataccta cgatgatgat
     1261 ctcgataacc tcctcgctca gatcggagat cagtacgctg atttgttcct cgctgctaag
     1321 aacctctctg atgctatcct cctcagtgat atcctcagag tgaacaccga gatcaccaag
     1381 gctccactct cagcttctat gatcaagaga tacgatgagc accaccagga tctcacactt
     1441 ctcaaggctc ttgttagaca gcagctccca gagaagtaca aagagatttt cttcgatcag
     1501 tctaagaacg gatacgctgg ttacatcgat ggtggtgcat ctcaagaaga gttctacaag
     1561 ttcatcaagc ctatcctcga gaagatggat ggaaccgagg aactcctcgt gaagctcaat
     1621 agagaggatc ttctcagaaa gcagaggacc ttcgataacg gatctatccc tcatcagatc
     1681 cacctcggag agttgcacgc tatccttaga aggcaagagg atttctaccc attcctcaag
     1741 gataacaggg aaaagattga gaagattctc accttcagaa tcccttacta cgtgggacct
     1801 ctcgctagag gaaactcaag attcgcttgg atgaccagaa agtctgagga aaccatcacc
     1861 ccttggaact tcgaagaggt ggtggataag ggtgctagtg ctcagtcttt catcgagagg
     1921 atgaccaact tcgataagaa ccttccaaac gagaaggtgc tccctaagca ctctttgctc
     1981 tacgagtact tcaccgtgta caacgagttg accaaggtta agtacgtgac cgagggaatg
     2041 aggaagcctg cttttttgtc aggtgagcaa aagaaggcta tcgttgatct cttgttcaag
     2101 accaacagaa aggtgaccgt gaagcagctc aaagaggatt acttcaagaa aatcgagtgc
     2161 ttcgattcag ttgagatttc tggtgttgag gataggttca acgcatctct cggaacctac
     2221 cacgatctcc tcaagatcat taaggataag gatttcttgg ataacgagga aaacgaggat
     2281 atcttggagg atatcgttct taccctcacc ctctttgaag atagagagat gattgaagaa
     2341 aggctcaaga cctacgctca tctcttcgat gataaggtga tgaagcagtt gaagagaaga
     2401 agatacactg gttggggaag gctctcaaga aagctcatta acggaatcag ggataagcag
     2461 tctggaaaga caatccttga tttcctcaag tctgatggat tcgctaacag aaacttcatg
     2521 cagctcatcc acgatgattc tctcaccttt aaagaggata tccagaaggc tcaggtttca
     2581 ggacagggtg atagtctcca tgagcatatc gctaacctcg ctggatctcc tgcaatcaag
     2641 aagggaatcc tccagactgt gaaggttgtg gatgagttgg tgaaggtgat gggaaggcat
     2701 aagcctgaga acatcgtgat cgaaatggct agagagaacc agaccactca gaagggacag
     2761 aagaactcta gggaaaggat gaagaggatc gaggaaggta tcaaagagct tggatctcag
     2821 atcctcaaag agcaccctgt tgagaacact cagctccaga atgagaagct ctacctctac
     2881 tacctccaga acggaaggga tatgtatgtg gatcaagagt tggatatcaa caggctctct
     2941 gattacgatg ttgatcatat cgtgccacag tcattcttga aggatgattc tatcgataac
     3001 aaggtgctca ccaggtctga taagaacagg ggtaagagtg ataacgtgcc aagtgaagag
     3061 gttgtgaaga aaatgaagaa ctattggagg cagctcctca acgctaagct catcactcag
     3121 agaaagttcg ataacttgac taaggctgag aggggaggac tctctgaatt ggataaggca
     3181 ggattcatca agaggcagct tgtggaaacc aggcagatca ctaagcacgt tgcacagatc
     3241 ctcgattcta ggatgaacac caagtacgat gagaacgata agttgatcag ggaagtgaag
     3301 gttatcaccc tcaagtcaaa gctcgtgtct gatttcagaa aggatttcca attctacaag
     3361 gtgagggaaa tcaacaacta ccaccacgct cacgatgctt accttaacgc tgttgttgga
     3421 accgctctca tcaagaagta tcctaagctc gagtcagagt tcgtgtacgg tgattacaag
     3481 gtgtacgatg tgaggaagat gatcgctaag tctgagcaag agatcggaaa ggctaccgct
     3541 aagtatttct tctactctaa catcatgaat ttcttcaaga ccgagattac cctcgctaac
     3601 ggtgagatca gaaagaggcc actcatcgag acaaacggtg aaacaggtga gatcgtgtgg
     3661 gataagggaa gggatttcgc taccgttaga aaggtgctct ctatgccaca ggtgaacatc
     3721 gttaagaaaa ccgaggtgca gaccggtgga ttctctaaag agtctatcct ccctaagagg
     3781 aactctgata agctcattgc taggaagaag gattgggacc ctaagaaata cggtggtttc
     3841 gattctccta ccgtggctta ctctgttctc gttgtggcta aggttgagaa gggaaagagt
     3901 aagaagctca agtctgttaa ggaacttctc ggaatcacta tcatggaaag gtcatctttc
     3961 gagaagaacc caatcgattt cctcgaggct aagggataca aagaggttaa gaaggatctc
     4021 atcatcaagc tcccaaagta ctcactcttc gaactcgaga acggtagaaa gaggatgctc
     4081 gcttctgctg gtgagcttca aaagggaaac gagcttgctc tcccatctaa gtacgttaac
     4141 tttctttacc tcgcttctca ctacgagaag ttgaagggat ctccagaaga taacgagcag
     4201 aagcaacttt tcgttgagca gcacaagcac tacttggatg agatcatcga gcagatctct
     4261 gagttctcta aaagggtgat cctcgctgat gcaaacctcg ataaggtgtt gtctgcttac
     4321 aacaagcaca gagataagcc tatcagggaa caggcagaga acatcatcca tctcttcacc
     4381 cttaccaacc tcggtgctcc tgctgctttc aagtacttcg atacaaccat cgataggaag
     4441 agatacacct ctaccaaaga agtgctcgat gctaccctca tccatcagtc tatcactgga
     4501 ctctacgaga ctaggatcga tctctcacag ctcggtggtg attcaagggc tgatcctaag
     4561 aagaagagga aggtttgaga cgcaggcctc ccagctttcg tccgtatcat cggtttcgac
     4621 aacgttcgtc aagttcaatg catcagtttc attgcccaca caccagaatc ctactaagtt
     4681 tgagtattat ggcattggaa aagctgtttt cttctatcat ttgttctgct tgtaatttac
     4741 tgtgttcttt cagtttttgt tttcggacat caaaatgcaa atggatggat aagagttaat
     4801 aaatgatatg gtccttttgt tcattctcaa attattatta tctgttgttt ttactttaat
     4861 gggttgaatt taagtaagaa aggaactaac agtgtgatat taaggtgcaa tgttagacat
     4921 ataaaacagt ctttcacctc tctttggtta tgtcttgaat tggtttgttt cttcacttat
     4981 ctgtgtaatc aagtttacta tgagtctatg atcaagtaat tatgcaatca agttaagtac
     5041 agtataggct ttcagcgaga ccggtggt
//
