LOCUS       sequence-91658-e       13943 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cas9 driven by double 35S, GFP for plant selection, I-PpoI site to 
            accept gRNA from pUC gRNA Shuttle.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13943)
  AUTHORS   Jacobs TB, LaFayette PR, Schmitz RJ, Parrott WA
  TITLE     Targeted genome modifications in soybean with CRISPR/Cas9.
  JOURNAL   BMC Biotechnol. 2015 Mar 12;15:16. doi: 10.1186/s12896-015-0131-2.
  PUBMED    25879861
REFERENCE   2  (bases 1 to 13943)
  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: "BMC 
            Biotechnol."; date: "2015-03-12"; volume: "15:16. doi"; pages: " 
            10.1186/s12896-015-0131-2"
COMMENT     Sequence Label: p201G Cas9
FEATURES             Location/Qualifiers
     source          1..13943
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1187..1816
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             2250..3317
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
                     AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
                     DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
                     VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
                     ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
                     RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
                     MRYRNLIEGEASAGS"
     rep_origin      3383..3577
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(3813..3835)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3921..4061
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3976..3995
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4076..4093)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4247..4835)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4327..4346)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4930..5745)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(4930..5745)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     5653..5672
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5737..5756
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     misc_feature    6038..6062
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     protein_bind    6239..6260
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6275..6280,6281..6298,6299..6305)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6313..6329
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     6318..6340
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6337..6353
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6337..6353
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      complement(6409..6661)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(join(6680..7393,7394..7396,7397..7399))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(6725..6746)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     7074..7093
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7333..7354
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     promoter        complement(7445..8121)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     promoter        8375..9051
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     CDS             9097..13200
                     /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             9097..13200
                     /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             9097..13200
                     /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             13213..13233
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      13300..13552
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(13594..13611)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(13594..13610)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13603..13625)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    13806..13830
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
ORIGIN
        1 agtactttga tccaacccct ccgctgctat agtgcagtcg gcttctgacg ttcagtgcag
       61 ccgtcttctg aaaacgacat gtcgcacaag tcctaagtta cgcgacaggc tgccgccctg
      121 cccttttcct ggcgttttct tgtcgcgtgt tttagtcgca taaagtagaa tacttgcgac
      181 tagaaccgga gacattacgc catgaacaag agcgccgccg ctggcctgct gggctatgcc
      241 cgcgtcagca ccgacgacca ggacttgacc aaccaacggg ccgaactgca cgcggccggc
      301 tgcaccaagc tgttttccga gaagatcacc ggcaccaggc gcgaccgccc ggagctggcc
      361 aggatgcttg accacctacg ccctggcgac gttgtgacag tgaccaggct agaccgcctg
      421 gcccgcagca cccgcgacct actggacatt gccgagcgca tccaggaggc cggcgcgggc
      481 ctgcgtagcc tggcagagcc gtgggccgac accaccacgc cggccggccg catggtgttg
      541 accgtgttcg ccggcattgc cgagttcgag cgttccctaa tcatcgaccg cacccggagc
      601 gggcgcgagg ccgccaaggc ccgaggcgtg aagtttggcc cccgccctac cctcaccccg
      661 gcacagatcg cgcacgcccg cgagctgatc gaccaggaag gccgcaccgt gaaagaggcg
      721 gctgcactgc ttggcgtgca tcgctcgacc ctgtaccgcg cacttgagcg cagcgaggaa
      781 gtgacgccca ccgaggccag gcggcgcggt gccttccgtg aggacgcatt gaccgaggcc
      841 gacgccctgg cggccgccga gaatgaacgc caagaggaac aagcatgaaa ccgcaccagg
      901 acggccagga cgaaccgttt ttcattaccg aagagatcga ggcggagatg atcgcggccg
      961 ggtacgtgtt cgagccgccc gcgcacgtct caaccgtgcg gctgcatgaa atcctggccg
     1021 gtttgtctga tgccaagctg gcggcctggc cggccagctt ggccgctgaa gaaaccgagc
     1081 gccgccgtct aaaaaggtga tgtgtatttg agtaaaacag cttgcgtcat gcggtcgctg
     1141 cgtatatgat gcgatgagta aataaacaaa tacgcaaggg gaacgcatga aggttatcgc
     1201 tgtacttaac cagaaaggcg ggtcaggcaa gacgaccatc gcaacccatc tagcccgcgc
     1261 cctgcaactc gccggggccg atgttctgtt agtcgattcc gatccccagg gcagtgcccg
     1321 cgattgggcg gccgtgcggg aagatcaacc gctaaccgtt gtcggcatcg accgcccgac
     1381 gattgaccgc gacgtgaagg ccatcggccg gcgcgacttc gtagtgatcg acggagcgcc
     1441 ccaggcggcg gacttggctg tgtccgcgat caaggcagcc gacttcgtgc tgattccggt
     1501 gcagccaagc ccttacgaca tatgggccac cgccgacctg gtggagctgg ttaagcagcg
     1561 cattgaggtc acggatggaa ggctacaagc ggcctttgtc gtgtcgcggg cgatcaaagg
     1621 cacgcgcatc ggcggtgagg ttgccgaggc gctggccggg tacgagctgc ccattcttga
     1681 gtcccgtatc acgcagcgcg tgagctaccc aggcactgcc gccgccggca caaccgttct
     1741 tgaatcagaa cccgagggcg acgctgcccg cgaggtccag gcgctggccg ctgaaattaa
     1801 atcaaaactc atttgagtta atgaggtaaa gagaaaatga gcaaaagcac aaacacgcta
     1861 agtgccggcc gtccgagcgc acgcagcagc aaggctgcaa cgttggccag cctggcagac
     1921 acgccagcca tgaagcgggt caactttcag ttgccggcgg aggatcacac caagctgaag
     1981 atgtacgcgg tacgccaagg caagaccatt accgagctgc tatctgaata catcgcgcag
     2041 ctaccagagt aaatgagcaa atgaataaat gagtagatga attttagcgg ctaaaggagg
     2101 cggcatggaa aatcaagaac aaccaggcac cgacgccgtg gaatgcccca tgtgtggagg
     2161 aacgggcggt tggccaggcg taagcggctg ggttgtctgc cggccctgca atggcactgg
     2221 aacccccaag cccgaggaat cggcgtgacg gtcgcaaacc atccggcccg gtacaaatcg
     2281 gcgcggcgct gggtgatgac ctggtggaga agttgaaggc cgcgcaggcc gcccagcggc
     2341 aacgcatcga ggcagaagca cgccccggtg aatcgtggca agcggccgct gatcgaatcc
     2401 gcaaagaatc ccggcaaccg ccggcagccg gtgcgccgtc gattaggaag ccgcccaagg
     2461 gcgacgagca accagatttt ttcgttccga tgctctatga cgtgggcacc cgcgatagtc
     2521 gcagcatcat ggacgtggcc gttttccgtc tgtcgaagcg tgaccgacga gctggcgagg
     2581 tgatccgcta cgagcttcca gacgggcacg tagaggtttc cgcagggccg gccggcatgg
     2641 ccagtgtgtg ggattacgac ctggtactga tggcggtttc ccatctaacc gaatccatga
     2701 accgataccg ggaagggaag ggagacaagc ccggccgcgt gttccgtcca cacgttgcgg
     2761 acgtactcaa gttctgccgg cgagccgatg gcggaaagca gaaagacgac ctggtagaaa
     2821 cctgcattcg gttaaacacc acgcacgttg ccatgcagcg tacgaagaag gccaagaacg
     2881 gccgcctggt gacggtatcc gagggtgaag ccttgattag ccgctacaag atcgtaaaga
     2941 gcgaaaccgg gcggccggag tacatcgaga tcgagctagc tgattggatg taccgcgaga
     3001 tcacagaagg caagaacccg gacgtgctga cggttcaccc cgattacttt ttgatcgatc
     3061 ccggcatcgg ccgttttctc taccgcctgg cacgccgcgc cgcaggcaag gcagaagcca
     3121 gatggttgtt caagacgatc tacgaacgca gtggcagcgc cggagagttc aagaagttct
     3181 gtttcaccgt gcgcaagctg atcgggtcaa atgacctgcc ggagtacgat ttgaaggagg
     3241 aggcggggca ggctggcccg atcctagtca tgcgctaccg caacctgatc gagggcgaag
     3301 catccgccgg ttcctaatgt acggagcaga tgctagggca aattgcccta gcaggggaaa
     3361 aaggtcgaaa aggtctcttt cctgtggata gcacgtacat tgggaaccca aagccgtaca
     3421 ttgggaaccg gaacccgtac attgggaacc caaagccgta cattgggaac cggtcacaca
     3481 tgtaagtgac tgatataaaa gagaaaaaag gcgatttttc cgcctaaaac tctttaaaac
     3541 ttattaaaac tcttaaaacc cgcctggcct gtgcataact gtctggccag cgcacagccg
     3601 aagagctgca aaaagcgcct acccttcggt cgctgcgctc cctacgcccc gccgcttcgc
     3661 gtcggcctat cgcggccgct ggccgctcaa aaatggctgg cctacggcca ggcaatctac
     3721 cagggcgcgg acaagccgcg ccgtcgccac tcgaccgccg gcgcccacat caaggcaccc
     3781 tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     3841 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     3901 ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat
     3961 actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     4021 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc
     4081 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     4141 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     4201 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4261 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4321 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4381 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4441 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4501 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4561 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4621 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4681 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4741 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4801 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4861 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg caggatcatg
     4921 aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg
     4981 attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag
     5041 gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc
     5101 aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg
     5161 agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc
     5221 aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat
     5281 tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac
     5341 aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga
     5401 atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag tggtgagtaa
     5461 ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt
     5521 cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg
     5581 tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga
     5641 ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt
     5701 taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac cccttgtatt
     5761 actgtttatg taagcagaca gttttattgt tcatgatctg gatcacaggc agcaacgctc
     5821 tgtcatcgtt acaatcaaca tgctaccctc cgcgagatca tccgtgtttc aaacccggca
     5881 gcttagttgc cgttcttccg aatagcatcg gtaacatgag caaagtctgc cgccttacaa
     5941 cggctctccc gctgacgccg tcccggactg atgggctgcc tgtatcgagt ggtgattttg
     6001 tgccgagctg ccggtcgggg agctgttggc tggctggtgg caggatatat tgtggtgtaa
     6061 acaaattgac gcttagacaa cttaataaca cattgcggac gtttttaatg tactgaatta
     6121 acgccgaatt gctctagcca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     6181 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     6241 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     6301 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gacatgatta
     6361 cgaattcctt gactagaggg taggcgcggc ttaattaaat caattcccga tctagtaaca
     6421 tagatgacac cgcgcgcgat aatttatcct agtttgcgcg ctatattttg ttttctatcg
     6481 cgtattaaat gtataattgc gggactctaa tcataaaaac ccatctcata aataacgtca
     6541 tgcattacat gttaattatt acatgcttaa cgtaattcaa cagaaattat atgataatca
     6601 tcgcaagacc ggcaacagga ttcaatctta agaaacttta ttgccaaatg tttgaacgat
     6661 ctgcagccgg gcggccgctt tacttgtaca gctcgtccat gccgtgagtg atcccggcgg
     6721 cggtcacgaa ctccagcagg accatgtgat cgcgcttctc gttggggtct ttgctcaggg
     6781 cggactgggt gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg
     6841 tgttctgctg gtagtggtcg gcgagctgca cgctgccgtc ctcgatgttg tggcggatct
     6901 tgaagttcac cttgatgccg ttcttctgct tgtcggccat gatatagacg ttgtggctgt
     6961 tgtagttgta ctccagcttg tgccccagga tgttgccgtc ctccttgaag tcgatgccct
     7021 tcagctcgat gcggttcacc agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt
     7081 agttgccgtc gtccttgaag aagatggtgc gctcctggac gtagccttcg ggcatggcgg
     7141 acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg gctgaagcac tgcacgccgt
     7201 aggtgaaggt ggtcacgagg gtgggccagg gcacgggcag cttgccggtg gtgcagatga
     7261 acttcagggt cagcttgccg taggtggcat cgccctcgcc ctcgccggac acgctgaact
     7321 tgtggccgtt tacgtcgccg tccagctcga ccaggatggg caccaccccg gtgaacagct
     7381 cctcgccctt gctcaccatg gatccggtcg agagagatag atttgtagag agagactggt
     7441 gatttcagcg tgtcctctcc aaatgaaatg aacttcctta tatagaggaa gggtcttgcg
     7501 aaggatagtg ggattgtgcg tcatccctta cgtcagtgga gatatcacat caatccactt
     7561 gctttgaaga cgtggttgga acgtcttctt tttccacgat gctcctcgtg ggtgggggtc
     7621 catctttggg accactgtcg gcagaggcat cttgaacgat agcctttcct ttatcgcaat
     7681 gatggcattt gtaggtgcca ccttcctttt ctactgtcct tttgatgaag tgacagatag
     7741 ctgggcaatg gaatccgagg aggtttcccg atattaccct ttgttgaaaa gtctcaatag
     7801 ccctttggtc ttctgagact gtatctttga tattcttgga gtagacgaga gtgtcgtgct
     7861 ccaccatgtt cacatcaatc cacttgcttt gaagacgtgg ttggaacgtc ttctttttcc
     7921 acgatgctcc tcgtgggtgg gggtccatct ttgggaccac tgtcggcaga ggcatcttga
     7981 acgatagcct ttcctttatc gcaatgatgg catttgtagg tgccaccttc cttttctact
     8041 gtccttttga tgaagtgaca gatagctggg caatggaatc cgaggaggtt tcccgatatt
     8101 accctttgtt gaaaagtctc aatagccctt tggtcttctg agactgtatc tttgatattc
     8161 ttggagtaga cgagagtgtc gtgctccacc atgttgggac tagtgagtcg tattacaatt
     8221 catgactctc ttaaggtagc caaaatcgat atctagggat aacagggtaa tcgatcacta
     8281 gaattcggcg cgccgggccc aacatggtgg agcacgacac tctcgtctac tccaagaata
     8341 tcaaagatac agtctcagaa gaccaaaggg ctattgagac ttttcaacaa agggtaatat
     8401 cgggaaacct cctcggattc cattgcccag ctatctgtca cttcatcaaa aggacagtag
     8461 aaaaggaagg tggcacctac aaatgccatc attgcgataa aggaaaggct atcgttcaag
     8521 atgcctctgc cgacagtggt cccaaagatg gacccccacc cacgaggagc atcgtggaaa
     8581 aagaagacgt tccaaccacg tcttcaaagc aagtggattg atgtgaacat ggtggagcac
     8641 gacactctcg tctactccaa gaatatcaaa gatacagtct cagaagacca aagggctatt
     8701 gagacttttc aacaaagggt aatatcggga aacctcctcg gattccattg cccagctatc
     8761 tgtcacttca tcaaaaggac agtagaaaag gaaggtggca cctacaaatg ccatcattgc
     8821 gataaaggaa aggctatcgt tcaagatgcc tctgccgaca gtggtcccaa agatggaccc
     8881 ccacccacga ggagcatcgt ggaaaaagaa gacgttccaa ccacgtcttc aaagcaagtg
     8941 gattgatgtg atatctccac tgacgtaagg gatgacgcac aatcccacta tccttcgcaa
     9001 gacccttcct ctatataagg aagttcattt catttggaga ggacacgctg aaatcaccag
     9061 tctctctcta caaatctatc tctctcgacc gctagcatgg acaagaagta ctccattggg
     9121 ctcgatatcg gcacaaacag cgtcggctgg gccgtcatta cggacgagta caaggtgccg
     9181 agcaaaaaat tcaaagttct gggcaatacc gatcgccaca gcataaagaa gaacctcatt
     9241 ggcgccctcc tgttcgactc cggggagacg gccgaagcca cgcggctcaa aagaacagca
     9301 cggcgcagat atacccgcag aaagaatcgg atctgctacc tgcaggagat ctttagtaat
     9361 gagatggcta aggtggatga ctctttcttc cataggctgg aggagtcctt tttggtggag
     9421 gaggataaaa agcacgagcg ccacccaatc tttggcaata tcgtggacga ggtggcgtac
     9481 catgaaaagt acccaaccat atatcatctg aggaagaagc ttgtagacag tactgataag
     9541 gctgacttgc ggttgatcta tctcgcgctg gcgcatatga tcaaatttcg gggacacttc
     9601 ctcatcgagg gggacctgaa cccagacaac agcgatgtcg acaaactctt tatccaactg
     9661 gttcagactt acaatcagct tttcgaagag aacccgatca acgcatccgg agttgacgcc
     9721 aaagcaatcc tgagcgctag gctgtccaaa tcccggcggc tcgaaaacct catcgcacag
     9781 ctccctgggg agaagaagaa cggcctgttt ggtaatctta tcgccctgtc actcgggctg
     9841 acccccaact ttaaatctaa cttcgacctg gccgaagatg ccaagcttca actgagcaaa
     9901 gacacctacg atgatgatct cgacaatctg ctggcccaga tcggcgacca gtacgcagac
     9961 ctttttttgg cggcaaagaa cctgtcagac gccattctgc tgagtgatat tctgcgagtg
    10021 aacacggaga tcaccaaagc tccgctgagc gctagtatga tcaagcgcta tgatgagcac
    10081 caccaagact tgactttgct gaaggccctt gtcagacagc aactgcctga gaagtacaag
    10141 gaaattttct tcgatcagtc taaaaatggc tacgccggat acattgacgg cggagcaagc
    10201 caggaggaat tttacaaatt tattaagccc atcttggaaa aaatggacgg caccgaggag
    10261 ctgctggtaa agcttaacag agaagatctg ttgcgcaaac agcgcacttt cgacaatgga
    10321 agcatccccc accagattca cctgggcgaa ctgcacgcta tcctcaggcg gcaagaggat
    10381 ttctacccct ttttgaaaga taacagggaa aagattgaga aaatcctcac atttcggata
    10441 ccctactatg taggccccct cgcccgggga aattccagat tcgcgtggat gactcgcaaa
    10501 tcagaagaga ccatcactcc ctggaacttc gaggaagtcg tggataaggg ggcctctgcc
    10561 cagtccttca tcgaaaggat gactaacttt gataaaaatc tgcctaacga aaaggtgctt
    10621 cctaaacact ctctgctgta cgagtacttc acagtttata acgagctcac caaggtcaaa
    10681 tacgtcacag aagggatgag aaagccagca ttcctgtctg gagagcagaa gaaagctatc
    10741 gtggacctcc tcttcaagac gaaccggaaa gttaccgtga aacagctcaa agaagactat
    10801 ttcaaaaaga ttgaatgttt cgactctgtt gaaatcagcg gagtggagga tcgcttcaac
    10861 gcatccctgg gaacgtatca cgatctcctg aaaatcatta aagacaagga cttcctggac
    10921 aatgaggaga acgaggacat tcttgaggac attgtcctca cccttacgtt gtttgaagat
    10981 agggagatga ttgaagaacg cttgaaaact tacgctcatc tcttcgacga caaagtcatg
    11041 aaacagctca agaggcgccg atatacagga tgggggcggc tgtcaagaaa actgatcaat
    11101 gggatccgag acaagcagag tggaaagaca atcctggatt ttcttaagtc cgatggattt
    11161 gccaaccgga acttcatgca gttgatccat gatgactctc tcacctttaa ggaggacatc
    11221 cagaaagcac aagtttctgg ccagggggac agtcttcacg agcacatcgc taatcttgca
    11281 ggtagcccag ctatcaaaaa gggaatactg cagaccgtta aggtcgtgga tgaactcgtc
    11341 aaagtaatgg gaaggcataa gcccgagaat atcgttatcg agatggcccg agagaaccaa
    11401 actacccaga agggacagaa gaacagtagg gaaaggatga agaggattga agagggtata
    11461 aaagaactgg ggtcccaaat ccttaaggaa cacccagttg aaaacaccca gcttcagaat
    11521 gagaagctct acctgtacta cctgcagaac ggcagggaca tgtacgtgga tcaggaactg
    11581 gacatcaatc ggctctccga ctacgacgtg gatcatatcg tgccccagtc ttttctcaaa
    11641 gatgattcta ttgataataa agtgttgaca agatccgata aaaatagagg gaagagtgat
    11701 aacgtcccct cagaagaagt tgtcaagaaa atgaaaaatt attggcggca gctgctgaac
    11761 gccaaactga tcacacaacg gaagttcgat aatctgacta aggctgaacg aggtggcctg
    11821 tctgagttgg ataaagccgg cttcatcaaa aggcagcttg ttgagacacg ccagatcacc
    11881 aagcacgtgg cccaaattct cgattcacgc atgaacacca agtacgatga aaatgacaaa
    11941 ctgattcgag aggtgaaagt tattactctg aagtctaagc tggtctcaga tttcagaaag
    12001 gactttcagt tttataaggt gagagagatc aacaattacc accatgcgca tgatgcctac
    12061 ctgaatgcag tggtaggcac tgcacttatc aaaaaatatc ccaagcttga atctgaattt
    12121 gtttacggag actataaagt gtacgatgtt aggaaaatga tcgcaaagtc tgagcaggaa
    12181 ataggcaagg ccaccgctaa gtacttcttt tacagcaata ttatgaattt tttcaagacc
    12241 gagattacac tggccaatgg agagattcgg aagcgaccac ttatcgaaac aaacggagaa
    12301 acaggagaaa tcgtgtggga caagggtagg gatttcgcga cagtccggaa ggtcctgtcc
    12361 atgccgcagg tgaacatcgt taaaaagacc gaagtacaga ccggaggctt ctccaaggaa
    12421 agtatcctcc cgaaaaggaa cagcgacaag ctgatcgcac gcaaaaaaga ttgggacccc
    12481 aagaaatacg gcggattcga ttctcctaca gtcgcttaca gtgtactggt tgtggccaaa
    12541 gtggagaaag ggaagtctaa aaaactcaaa agcgtcaagg aactgctggg catcacaatc
    12601 atggagcgat caagcttcga aaaaaacccc atcgactttc tcgaggcgaa aggatataaa
    12661 gaggtcaaaa aagacctcat cattaagctt cccaagtact ctctctttga gcttgaaaac
    12721 ggccggaaac gaatgctcgc tagtgcgggc gagctgcaga aaggtaacga gctggcactg
    12781 ccctctaaat acgttaattt cttgtatctg gccagccact atgaaaagct caaagggtct
    12841 cccgaagata atgagcagaa gcagctgttc gtggaacaac acaaacacta ccttgatgag
    12901 atcatcgagc aaataagcga attctccaaa agagtgatcc tcgccgacgc taacctcgat
    12961 aaggtgcttt ctgcttacaa taagcacagg gataagccca tcagggagca ggcagaaaac
    13021 attatccact tgtttactct gaccaacttg ggcgcgcctg cagccttcaa gtacttcgac
    13081 accaccatag acagaaagcg gtacacctct acaaaggagg tcctggacgc cacactgatt
    13141 catcagtcaa ttacggggct ctatgaaaca agaatcgacc tctctcagct cggtggagac
    13201 agcagggctg accccaagaa gaagaggaag gtgtgaccgc ggatatctgc agaagcttac
    13261 gcgtcgacgt ctcgaggccg gccgagctcg aatttccccg atcgttcaaa catttggcaa
    13321 taaagtttct taagattgaa tcctgttgcc ggtcttgcga tgattatcat ataatttctg
    13381 ttgaattacg ttaagcatgt aataattaac atgtaatgca tgacgttatt tatgagatgg
    13441 gtttttatga ttagagtccc gcaattatac atttaatacg cgatagaaaa caaaatatag
    13501 cgcgcaaact aggataaatt atcgcgcgcg gtgtcatcta tgttactaga tcgggaattg
    13561 atatcgtagg gataacaggg taataagctt ggcactggcc gtcgttttac aacgtcgtga
    13621 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
    13681 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
    13741 tggcgaatga gcttgagctt ggatcagatt gtcgtttccc gccttcagtt taaactatca
    13801 gtgtttgaca ggatatattg gcgggtaaac ctaagagaaa agagcgttta ttagaataac
    13861 ggatatttaa aagggcgtga aaaggtttat ccgttcgtcc atttgtatgt gcatgccaac
    13921 cacagggttc ccctcgggat caa
//
