LOCUS       sequence-181756-        7480 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Module A, Promoter: EC1.2, Gene: AtCas9, Terminator: HSP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7480)
  AUTHORS   Cermak T, Curtin SJ, Gil-Humanes J, Cegan R, Kono TJY, Konecna E, 
            Belanto JJ, Starker CG, Mathre JW, Greenstein RL, Voytas DF
  TITLE     A multi-purpose toolkit to enable advanced genome engineering in 
            plants.
  JOURNAL   Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 
            10.1105/tpc.16.00922.
  PUBMED    28522548
REFERENCE   2  (bases 1 to 7480)
  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 Cell.
            2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922."
COMMENT     Sequence Label: pMOD_A0108
FEATURES             Location/Qualifiers
     source          1..7480
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      80..668
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     569..588
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     822..839
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             1914..6017
                     /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             1914..6017
                     /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             6030..6050
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      6063..6311
                     /label=HSP terminator
                     /note="efficient transcription terminator from the 
                     Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya 
                     et al., 2010)"
     promoter        6388..6492
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gccttgacgg caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
       61 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
      121 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ctaactcttt
      181 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
      241 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
      301 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
      361 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgctcacagc
      421 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
      481 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
      541 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
      601 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
      661 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
      721 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
      781 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
      841 aagcggaaga gcgcccacct gctcatgatc ggcgcgccac tttgataaat gttcctcgct
      901 gacgtaagaa gacattagta atggttataa tatatagctt tctatgaatg tatggtgaga
      961 aaatgtctgt tcactgattt tgagtttgga ataaaagcat ttgcgtttgg tttatcattg
     1021 cgtttataca aggacagaga tccactgagc tggaatagct taaaaccatt atcagaacaa
     1081 aataaaccat tttttgttaa gaatcagagc atagtaaaca acagaaacaa cctaagagag
     1141 gtaacttgtc caagaagata gctaattata tctattttat aaaagttatc atagtttgta
     1201 agtcacaaaa gatgcaaata acagagaaac taggagactt gagaatatac attcttgtat
     1261 atttgtattc gagattgtga aaatttgacc ataagtttaa attcttaaaa agatatatct
     1321 gatctagatg atggttatag actgtaattt taccacatgt ttaatgatgg atagtgacac
     1381 acatgacaca tcgacaacac tatagcatct tatttagatt acaacatgaa atttttctgt
     1441 aatacatgtc tttgtacata atttaaaagt aattcctaag aaatatattt atacaaggag
     1501 tttaaagaaa acatagcata aagttcaatg agtagtaaaa accatataca gtatatagca
     1561 taaagttcaa tgagtttatt acaaaagcat tggttcactt tctgtaacac gacgttaaac
     1621 cttcgtctcc aataggagcg ctactgattc aacatgccaa tatatactaa atacgtttct
     1681 acagtcaaat gctttaacgt ttcatgatta agtgactatt taccgtcaat cctttcccat
     1741 tcctcccact aatccaactt tttaattact cttaaatcac cactaagctt cgaatccatc
     1801 caaaaccaca atataaaaac agaactctcg taactcaatc atcgcaaaac aaaacaaaac
     1861 aaaacaaaaa ccccaaaaag aaagaataac ctgcaggtag atcgctcgtc gacatggata
     1921 agaagtactc tatcggactc gatatcggaa ctaactctgt gggatgggct gtgatcaccg
     1981 atgagtacaa ggtgccatct aagaagttca aggttctcgg aaacaccgat aggcactcta
     2041 tcaagaaaaa ccttatcggt gctctcctct tcgattctgg tgaaactgct gaggctacca
     2101 gactcaagag aaccgctaga agaaggtaca ccagaagaaa gaacaggatc tgctacctcc
     2161 aagagatctt ctctaacgag atggctaaag tggatgattc attcttccac aggctcgaag
     2221 agtcattcct cgtggaagaa gataagaagc acgagaggca ccctatcttc ggaaacatcg
     2281 ttgatgaggt ggcataccac gagaagtacc ctactatcta ccacctcaga aagaagctcg
     2341 ttgattctac tgataaggct gatctcaggc tcatctacct cgctctcgct cacatgatca
     2401 agttcagagg acacttcctc atcgagggtg atctcaaccc tgataactct gatgtggata
     2461 agttgttcat ccagctcgtg cagacctaca accagctttt cgaagagaac cctatcaacg
     2521 cttcaggtgt ggatgctaag gctatcctct ctgctaggct ctctaagtca agaaggcttg
     2581 agaacctcat tgctcagctc cctggtgaga agaagaacgg acttttcgga aacttgatcg
     2641 ctctctctct cggactcacc cctaacttca agtctaactt cgatctcgct gaggatgcaa
     2701 agctccagct ctcaaaggat acctacgatg atgatctcga taacctcctc gctcagatcg
     2761 gagatcagta cgctgatttg ttcctcgctg ctaagaacct ctctgatgct atcctcctca
     2821 gtgatatcct cagagtgaac accgagatca ccaaggctcc actctcagct tctatgatca
     2881 agagatacga tgagcaccac caggatctca cacttctcaa ggctcttgtt agacagcagc
     2941 tcccagagaa gtacaaagag attttcttcg atcagtctaa gaacggatac gctggttaca
     3001 tcgatggtgg tgcatctcaa gaagagttct acaagttcat caagcctatc ctcgagaaga
     3061 tggatggaac cgaggaactc ctcgtgaagc tcaatagaga ggatcttctc agaaagcaga
     3121 ggaccttcga taacggatct atccctcatc agatccacct cggagagttg cacgctatcc
     3181 ttagaaggca agaggatttc tacccattcc tcaaggataa cagggaaaag attgagaaga
     3241 ttctcacctt cagaatccct tactacgtgg gacctctcgc tagaggaaac tcaagattcg
     3301 cttggatgac cagaaagtct gaggaaacca tcaccccttg gaacttcgaa gaggtggtgg
     3361 ataagggtgc tagtgctcag tctttcatcg agaggatgac caacttcgat aagaaccttc
     3421 caaacgagaa ggtgctccct aagcactctt tgctctacga gtacttcacc gtgtacaacg
     3481 agttgaccaa ggttaagtac gtgaccgagg gaatgaggaa gcctgctttt ttgtcaggtg
     3541 agcaaaagaa ggctatcgtt gatctcttgt tcaagaccaa cagaaaggtg accgtgaagc
     3601 agctcaaaga ggattacttc aagaaaatcg agtgcttcga ttcagttgag atttctggtg
     3661 ttgaggatag gttcaacgca tctctcggaa cctaccacga tctcctcaag atcattaagg
     3721 ataaggattt cttggataac gaggaaaacg aggatatctt ggaggatatc gttcttaccc
     3781 tcaccctctt tgaagataga gagatgattg aagaaaggct caagacctac gctcatctct
     3841 tcgatgataa ggtgatgaag cagttgaaga gaagaagata cactggttgg ggaaggctct
     3901 caagaaagct cattaacgga atcagggata agcagtctgg aaagacaatc cttgatttcc
     3961 tcaagtctga tggattcgct aacagaaact tcatgcagct catccacgat gattctctca
     4021 cctttaaaga ggatatccag aaggctcagg tttcaggaca gggtgatagt ctccatgagc
     4081 atatcgctaa cctcgctgga tctcctgcaa tcaagaaggg aatcctccag actgtgaagg
     4141 ttgtggatga gttggtgaag gtgatgggaa ggcataagcc tgagaacatc gtgatcgaaa
     4201 tggctagaga gaaccagacc actcagaagg gacagaagaa ctctagggaa aggatgaaga
     4261 ggatcgagga aggtatcaaa gagcttggat ctcagatcct caaagagcac cctgttgaga
     4321 acactcagct ccagaatgag aagctctacc tctactacct ccagaacgga agggatatgt
     4381 atgtggatca agagttggat atcaacaggc tctctgatta cgatgttgat catatcgtgc
     4441 cacagtcatt cttgaaggat gattctatcg ataacaaggt gctcaccagg tctgataaga
     4501 acaggggtaa gagtgataac gtgccaagtg aagaggttgt gaagaaaatg aagaactatt
     4561 ggaggcagct cctcaacgct aagctcatca ctcagagaaa gttcgataac ttgactaagg
     4621 ctgagagggg aggactctct gaattggata aggcaggatt catcaagagg cagcttgtgg
     4681 aaaccaggca gatcactaag cacgttgcac agatcctcga ttctaggatg aacaccaagt
     4741 acgatgagaa cgataagttg atcagggaag tgaaggttat caccctcaag tcaaagctcg
     4801 tgtctgattt cagaaaggat ttccaattct acaaggtgag ggaaatcaac aactaccacc
     4861 acgctcacga tgcttacctt aacgctgttg ttggaaccgc tctcatcaag aagtatccta
     4921 agctcgagtc agagttcgtg tacggtgatt acaaggtgta cgatgtgagg aagatgatcg
     4981 ctaagtctga gcaagagatc ggaaaggcta ccgctaagta tttcttctac tctaacatca
     5041 tgaatttctt caagaccgag attaccctcg ctaacggtga gatcagaaag aggccactca
     5101 tcgagacaaa cggtgaaaca ggtgagatcg tgtgggataa gggaagggat ttcgctaccg
     5161 ttagaaaggt gctctctatg ccacaggtga acatcgttaa gaaaaccgag gtgcagaccg
     5221 gtggattctc taaagagtct atcctcccta agaggaactc tgataagctc attgctagga
     5281 agaaggattg ggaccctaag aaatacggtg gtttcgattc tcctaccgtg gcttactctg
     5341 ttctcgttgt ggctaaggtt gagaagggaa agagtaagaa gctcaagtct gttaaggaac
     5401 ttctcggaat cactatcatg gaaaggtcat ctttcgagaa gaacccaatc gatttcctcg
     5461 aggctaaggg atacaaagag gttaagaagg atctcatcat caagctccca aagtactcac
     5521 tcttcgaact cgagaacggt agaaagagga tgctcgcttc tgctggtgag cttcaaaagg
     5581 gaaacgagct tgctctccca tctaagtacg ttaactttct ttacctcgct tctcactacg
     5641 agaagttgaa gggatctcca gaagataacg agcagaagca acttttcgtt gagcagcaca
     5701 agcactactt ggatgagatc atcgagcaga tctctgagtt ctctaaaagg gtgatcctcg
     5761 ctgatgcaaa cctcgataag gtgttgtctg cttacaacaa gcacagagat aagcctatca
     5821 gggaacaggc agagaacatc atccatctct tcacccttac caacctcggt gctcctgctg
     5881 ctttcaagta cttcgataca accatcgata ggaagagata cacctctacc aaagaagtgc
     5941 tcgatgctac cctcatccat cagtctatca ctggactcta cgagactagg atcgatctct
     6001 cacagctcgg tggtgattca agggctgatc ctaagaagaa gaggaaggtt tgacgtcgac
     6061 gatatgaaga tgaagatgaa atatttggtg tgtcaaataa aaagcttgtg tgcttaagtt
     6121 tgtgtttttt tcttggcttg ttgtgttatg aatttgtggc tttttctaat attaaatgaa
     6181 tgtaagatca cattataatg aataaacaaa tgtttctata atccattgtg aatgttttgt
     6241 tggatctctt ctgcagcata taactactgt atgtgctatg gtatggacta tggaatatga
     6301 ttaaagataa gccagagctc tggtgacgga cgtgggcagg tgtgatgaaa agccttaggg
     6361 tcaggtggca cttttcgggg aaatgtgcga tgaaccccta tttgtttatt tttctaaata
     6421 cattcaaata tgtatccgct aatgagacaa taaccctgat aaatgcttca attatattga
     6481 aaaaggaaga gtatgagtat ccaacatttc cgtgtcgccc ttattccctt ttttgcggcc
     6541 ttctgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     6601 cagttgggtg cccgagtggg ttacattgaa ctggatctca acagcggcaa aatcttagag
     6661 agttttcgcc ccgaagaacg cttcccaatg atgagcactt tcaaagttct gctatgtggc
     6721 gcggtgttat cccgtattga tgctgggcaa gagcaactcg gtcgccgcat acactattct
     6781 cagaatgact tggttgagta ttcaccagtc acagaaaagc atcttacgga tggcatgaca
     6841 gtaagagaat tatgtagtgc tgccataacc atgagtgata acactgcggc caacttactt
     6901 ctgacaacga ttggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     6961 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     7021 gacaccacga tgcctgtagc aatggcaaca actctccgca aactattaac tggcgaacta
     7081 cttactcttg cttcacgcca gcaactcatt gactggatgg aggcggacaa agttgcagga
     7141 ccacttctgc gctcggcact tccggctggc tggtttattg ctgataaatc tggagccggt
     7201 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     7261 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     7321 gagataggtg cctcactgat taagcattgg taactgtcag acgaagggaa ataaatagta
     7381 gcccgcctga tgtgcgggct tttttttggt ggtggttacc agtgtcgctt agctaccgta
     7441 ggtcctctta ggcggtccga ttagcgacat agtcatcaat
//
