LOCUS       sequence-74310-e       10591 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 2 construct.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10591)
  AUTHORS   Belhaj K, Chaparro-Garcia A, Kamoun S, Nekrasov V
  TITLE     Plant genome editing made easy: targeted mutagenesis in model and 
            crop plants using the CRISPR/Cas system.
  JOURNAL   Plant Methods. 2013 Oct 11;9(1):39.
  PUBMED    24112467
REFERENCE   2  (bases 1 to 10591)
  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 
            Methods."; date: "2013-10-11"; volume: "9(1)"; pages: "39"
COMMENT     Sequence Label: 
            pAGM4723::AtU6p::sgRNA2-2x35S-5′UTR::Cas9::NOST-AtU6p::sgRNA1
FEATURES             Location/Qualifiers
     source          1..10591
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        22..96
                     /label=AtU6 promoter
                     /note="consensus promoter from the three Arabidopsis U6 
                     snRNA genes (Waibel and Filipowicz, 1990; Nekrasov et al., 
                     2013)"
     misc_RNA        117..192
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(204..228)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        662..1007
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     950..970
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             1098..5201
                     /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             1098..5201
                     /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             1098..5201
                     /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             5214..5234
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      5250..5502
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        5529..5603
                     /label=AtU6 promoter
                     /note="consensus promoter from the three Arabidopsis U6 
                     snRNA genes (Waibel and Filipowicz, 1990; Nekrasov et al., 
                     2013)"
     misc_RNA        5624..5699
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(5711..5735)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    5804..5828
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             6108..6737
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             7171..8238
                     /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      8304..8498
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     promoter        8706..8810
                     /gene="bla"
                     /label=AmpR promoter
     CDS             8811..9605
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      9776..10364
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10265..10284
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    10500..10524
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 gtgccgaatt cggatccgga gtgatcaaaa gtcccacatc gatcaggtga tatatagcag
       61 cttagtttat ataatgatag agtcgacata gcgattgtca agatgtttgc ttgcaagttt
      121 tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca
      181 ccgagtcggt gctttttttc tagacccagc tttcttgtac aaagttggca ttacgctgca
      241 agaattcaag cttggaggtc aacatggtgg agcacgacac tctggtctac tccaaaaatg
      301 tcaaagatac agtctcagaa gatcaaaggg ctattgagac ttttcaacaa aggataattt
      361 cgggaaacct cctcggattc cattgcccag ctatctgtca cttcatcgaa aggacagtag
      421 aaaaggaagg tggctcctac aaatgccatc attgcgataa aggaaaggct atcattcaag
      481 atctctctgc cgacagtggt cccaaagatg gacccccacc cacgaggagc atcgtggaaa
      541 aagaagaggt tccaaccacg tctacaaagc aagtggattg atgtgataac atggtggagc
      601 acgacactct ggtctactcc aaaaatgtca aagatacagt ctcagaagat caaagggcta
      661 ttgagacttt tcaacaaagg ataatttcgg gaaacctcct cggattccat tgcccagcta
      721 tctgtcactt catcgaaagg acagtagaaa aggaaggtgg ctcctacaaa tgccatcatt
      781 gcgataaagg aaaggctatc attcaagatc tctctgccga cagtggtccc aaagatggac
      841 ccccacccac gaggagcatc gtggaaaaag aagaggttcc aaccacgtct acaaagcaag
      901 tggattgatg tgacatctcc actgacgtaa gggatgacgc acaatcccac tatccttcgc
      961 aagacccttc ctctatataa ggaagttcat ttcatttgga gaggacacgc tcgagtataa
     1021 gagctcattt ttacaacaat taccaacaac aacaaacaac aaacaacatt acaattacat
     1081 ttacaattat cgatacaatg gacaagaagt actccattgg gctcgatatc ggcacaaaca
     1141 gcgtcggctg ggccgtcatt acggacgagt acaaggtgcc gagcaaaaaa ttcaaagttc
     1201 tgggcaatac cgatcgccac agcataaaga agaacctcat tggcgccctc ctgttcgact
     1261 ccggggagac ggccgaagcc acgcggctca aaagaacagc acggcgcaga tatacccgca
     1321 gaaagaatcg gatctgctac ctgcaggaga tctttagtaa tgagatggct aaggtggatg
     1381 actctttctt ccataggctg gaggagtcct ttttggtgga ggaggataaa aagcacgagc
     1441 gccacccaat ctttggcaat atcgtggacg aggtggcgta ccatgaaaag tacccaacca
     1501 tatatcatct gaggaagaag cttgtagaca gtactgataa ggctgacttg cggttgatct
     1561 atctcgcgct ggcgcatatg atcaaatttc ggggacactt cctcatcgag ggggacctga
     1621 acccagacaa cagcgatgtc gacaaactct ttatccaact ggttcagact tacaatcagc
     1681 ttttcgaaga gaacccgatc aacgcatccg gagttgacgc caaagcaatc ctgagcgcta
     1741 ggctgtccaa atcccggcgg ctcgaaaacc tcatcgcaca gctccctggg gagaagaaga
     1801 acggcctgtt tggtaatctt atcgccctgt cactcgggct gacccccaac tttaaatcta
     1861 acttcgacct ggccgaagat gccaagcttc aactgagcaa agacacctac gatgatgatc
     1921 tcgacaatct gctggcccag atcggcgacc agtacgcaga cctttttttg gcggcaaaga
     1981 acctgtcaga cgccattctg ctgagtgata ttctgcgagt gaacacggag atcaccaaag
     2041 ctccgctgag cgctagtatg atcaagcgct atgatgagca ccaccaagac ttgactttgc
     2101 tgaaggccct tgtcagacag caactgcctg agaagtacaa ggaaattttc ttcgatcagt
     2161 ctaaaaatgg ctacgccgga tacattgacg gcggagcaag ccaggaggaa ttttacaaat
     2221 ttattaagcc catcttggaa aaaatggacg gcaccgagga gctgctggta aagcttaaca
     2281 gagaagatct gttgcgcaaa cagcgcactt tcgacaatgg aagcatcccc caccagattc
     2341 acctgggcga actgcacgct atcctcaggc ggcaagagga tttctacccc tttttgaaag
     2401 ataacaggga aaagattgag aaaatcctca catttcggat accctactat gtaggccccc
     2461 tcgcccgggg aaattccaga ttcgcgtgga tgactcgcaa atcagaagag actatcactc
     2521 cctggaactt cgaggaagtc gtggataagg gggcctctgc ccagtccttc atcgaaagga
     2581 tgactaactt tgataaaaat ctgcctaacg aaaaggtgct tcctaaacac tctctgctgt
     2641 acgagtactt cacagtttat aacgagctca ccaaggtcaa atacgtcaca gaagggatga
     2701 gaaagccagc attcctgtct ggagagcaga agaaagctat cgtggacctc ctcttcaaga
     2761 cgaaccggaa agttaccgtg aaacagctca aagaagatta tttcaaaaag attgaatgtt
     2821 tcgactctgt tgaaatcagc ggagtggagg atcgcttcaa cgcatccctg ggaacgtatc
     2881 acgatctcct gaaaatcatt aaagacaagg acttcctgga caatgaggag aacgaggaca
     2941 ttcttgagga cattgtcctc acccttacgt tgtttgaaga tagggagatg attgaagaac
     3001 gcttgaaaac ttacgctcat ctcttcgacg acaaagtcat gaaacagctc aagaggcgcc
     3061 gatatacagg atgggggcgg ctgtcaagaa aactgatcaa tgggatccga gacaagcaga
     3121 gtggaaagac aatcctggat tttcttaagt ccgatggatt tgccaaccgg aacttcatgc
     3181 agttgatcca tgatgactct ctcaccttta aggaggacat ccagaaagca caagtttctg
     3241 gccaggggga cagtctccac gagcacatcg ctaatcttgc aggtagccca gctatcaaaa
     3301 agggaatact gcagaccgtt aaggtcgtgg atgaactcgt caaagtaatg ggaaggcata
     3361 agcccgagaa tatcgttatc gagatggccc gagagaacca aactacccag aagggacaga
     3421 agaacagtag ggaaaggatg aagaggattg aagagggtat aaaagaactg gggtcccaaa
     3481 tccttaagga acacccagtt gaaaacaccc agcttcagaa tgagaagctc tacctgtact
     3541 acctgcagaa cggcagggac atgtacgtgg atcaggaact ggacatcaat cggctctccg
     3601 actacgacgt ggatcatatc gtgccccagt cttttctcaa agatgattct attgataata
     3661 aagtgttgac aagatccgat aaaaatagag ggaagagtga taacgtcccc tcagaagaag
     3721 ttgtcaagaa aatgaaaaat tattggcggc agctgctgaa cgccaaactg atcacacaac
     3781 ggaagttcga taatctgact aaggctgaac gaggtggcct gtctgagttg gataaagccg
     3841 gcttcatcaa aaggcagctt gttgagacac gccagatcac caagcacgtg gcccaaattc
     3901 tcgattcacg catgaacacc aagtacgatg aaaatgacaa actgattcga gaggtgaaag
     3961 ttattactct gaagtctaag ctggtttcag atttcagaaa ggactttcag ttttataagg
     4021 tgagagagat caacaattac caccatgcgc atgatgccta cctgaatgca gtggtaggca
     4081 ctgcacttat caaaaaatat cccaagcttg aatctgaatt tgtttacgga gactataaag
     4141 tgtacgatgt taggaaaatg atcgcaaagt ctgagcagga aataggcaag gccaccgcta
     4201 agtacttctt ttacagcaat attatgaatt ttttcaagac cgagattaca ctggccaatg
     4261 gagagattcg gaagcgacca cttatcgaaa caaacggaga aacaggagaa atcgtgtggg
     4321 acaagggtag ggatttcgcg acagtccgga aggtcctgtc catgccgcag gtgaacatcg
     4381 ttaaaaagac cgaagtacag accggaggct tctccaagga aagtatcctc ccgaaaagga
     4441 acagcgacaa gctgatcgca cgcaaaaaag attgggaccc caagaaatac ggcggattcg
     4501 attctcctac agtcgcttac agtgtactgg ttgtggccaa agtggagaaa gggaagtcta
     4561 aaaaactcaa aagcgtcaag gaactgctgg gcatcacaat catggagcga tcaagcttcg
     4621 aaaaaaaccc catcgacttt ctcgaggcga aaggatataa agaggtcaaa aaagacctca
     4681 tcattaagct tcccaagtac tctctctttg agcttgaaaa cggccggaaa cgaatgctcg
     4741 ctagtgcggg cgagctgcag aaaggtaacg agctggcact gccctctaaa tacgttaatt
     4801 tcttgtatct ggccagccac tatgaaaagc tcaaaggatc tcccgaagat aatgagcaga
     4861 agcagctgtt cgtggaacaa cacaaacact accttgatga gatcatcgag caaataagcg
     4921 aattctccaa aagagtgatc ctcgccgacg ctaacctcga taaggtgctt tctgcttaca
     4981 ataagcacag ggataagccc atcagggagc aggcagaaaa cattatccac ttgtttactc
     5041 tgaccaactt gggcgcgcct gcagccttca agtacttcga caccaccata gacagaaagc
     5101 ggtacacctc tacaaaggag gtcctggacg ccacactgat tcatcagtca attacggggc
     5161 tctatgaaac aagaatcgac ctctctcagc tcggtggaga cagcagggct gaccccaaga
     5221 agaagaggaa ggtgtgagct tgtcaagcag atcgttcaaa catttggcaa taaagtttct
     5281 taagattgaa tcctgttgcc ggtcttgcga tgattatcat ataatttctg ttgaattacg
     5341 ttaagcatgt aataattaac atgtaatgca tgacgttatt tatgagatgg gtttttatga
     5401 ttagagtccc gcaattatac atttaatacg cgatagaaaa caaaatatag cgcgcaaact
     5461 aggataaatt atcgcgcgcg gtgtcatcta tgttactaga tcgacgctac tagaattcga
     5521 gctcggagtg atcaaaagtc ccacatcgat caggtgatat atagcagctt agtttatata
     5581 atgatagagt cgacatagcg attgccgtta atttgagagt ccagttttag agctagaaat
     5641 agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct
     5701 ttttttctag acccagcttt cttgtacaaa gttggcatta cgctttacga ggatgcacat
     5761 gtgaccgagg gacacgaagt gatccgttta aactatcagt gtttgacagg atatattggc
     5821 gggtaaacct aagagaaaag agcgtttatt agaataatcg gatatttaaa agggcgtgaa
     5881 aaggtttatc cgttcgtcca tttgtatgtg ccagccgtgc ggctgcatga aatcctggcc
     5941 ggtttgtctg atgccaagct ggcggcctgg ccggccagct tggccgctga agaaaccgag
     6001 cgccgccgtc taaaaaggtg atgtgtattt gagtaaaaca gcttgcgtca tgcggtcgct
     6061 gcgtatatga tgcgatgagt aaataaacaa atacgcaagg ggaacgcatg aaggttatcg
     6121 ctgtacttaa ccagaaaggc gggtcaggca agacgaccat cgcaacccat ctagcccgcg
     6181 ccctgcaact cgccggggcc gatgttctgt tagtcgattc cgatccccag ggcagtgccc
     6241 gcgattgggc ggccgtgcgg gaagatcaac cgctaaccgt tgtcggcatc gaccgcccga
     6301 cgattgaccg cgacgtgaag gccatcggcc ggcgcgactt cgtagtgatc gacggagcgc
     6361 cccaggcggc ggacttggct gtgtccgcga tcaaggcagc cgacttcgtg ctgattccgg
     6421 tgcagccaag cccttacgac atatgggcca ccgccgacct ggtggagctg gttaagcagc
     6481 gcattgaggt cacggatgga aggctacaag cggcctttgt cgtgtcgcgg gcgatcaaag
     6541 gcacgcgcat cggcggtgag gttgccgagg cgctggccgg gtacgagctg cccattcttg
     6601 agtcccgtat cacgcagcgc gtgagctacc caggcactgc cgccgccggc acaaccgttc
     6661 ttgaatcaga acccgagggc gacgctgccc gcgaggtcca ggcgctggcc gctgaaatta
     6721 aatcaaaact catttgagtt aatgaggtaa agagaaaatg agcaaaagca caaacacgct
     6781 aagtgccggc cgtccgagcg cacgcagcag caaggctgca acgttggcca gcctggcaga
     6841 cacgccagcc atgaagcggg tcaactttca gttgccggcg gaggatcaca ccaagctgaa
     6901 gatgtacgcg gtacgccaag gcaagaccat taccgagctg ctatctgaat acatcgcgca
     6961 gctaccagag taaatgagca aatgaataaa tgagtagatg aattttagcg gctaaaggag
     7021 gcggcatgga aaatcaagaa caaccaggca ccgacgccgt ggaatgcccc atgtgtggag
     7081 gaacgggcgg ttggccaggc gtaagcggct gggttgtctg ccggccctgc aatggcactg
     7141 gaacccccaa gcccgaggaa tcggcgtgac ggtcgcaaac catccggccc ggtacaaatc
     7201 ggcgcggcgc tgggtgatga cctggtggag aagttgaagg ccgcgcaggc cgcccagcgg
     7261 caacgcatcg aggcagaagc acgccccggt gaatcgtggc aagcggccgc tgatcgaatc
     7321 cgcaaagaat cccggcaacc gccggcagcc ggtgcgccgt cgattaggaa gccgcccaag
     7381 ggcgacgagc aaccagattt tttcgttccg atgctctatg acgtgggcac ccgcgatagt
     7441 cgcagcatca tggacgtggc cgttttccgt ctgtcgaagc gtgaccgacg agctggcgag
     7501 gtgatccgct acgagcttcc agacgggcac gtagaggttt ccgcagggcc ggccggcatg
     7561 gccagtgtgt gggattacga cctggtactg atggcggttt cccatctaac cgaatccatg
     7621 aaccgatacc gggaagggaa gggagacaag cccggccgcg tgttccgtcc acacgttgcg
     7681 gacgtactca agttctgccg gcgagccgat ggcggaaagc agaaagacga cctggtagaa
     7741 acctgcattc ggttaaacac cacgcacgtt gccatgcagc gtacgaagaa ggccaagaac
     7801 ggccgcctgg tgacggtatc cgagggtgaa gccttgatta gccgctacaa gatcgtaaag
     7861 agcgaaaccg ggcggccgga gtacatcgag atcgagctag ctgattggat gtaccgcgag
     7921 atcacagaag gcaagaaccc ggacgtgctg acggttcacc ccgattactt tttgatcgat
     7981 cccggcatcg gccgttttct ctaccgcctg gcacgccgcg ccgcaggcaa ggcagaagcc
     8041 agatggttgt tcaagacgat ctacgaacgc agtggcagcg ccggagagtt caagaagttc
     8101 tgtttcaccg tgcgcaagct gatcgggtca aatgacctgc cggagtacga tttgaaggag
     8161 gaggcggggc aggctggccc gatcctagtc atgcgctacc gcaacctgat cgagggcgaa
     8221 gcatccgccg gttcctaatg tacggagcag atgctagggc aaattgccct agcaggggaa
     8281 aaaggtcgaa aaagcttctt tcctgtggat agcacgtaca ttgggaaccc aaagccgtac
     8341 attgggaacc ggaacccgta cattgggaac ccaaagccgt acattgggaa ccggtcacac
     8401 atgtaagtga ctgatataaa agagaaaaaa ggcgattttt ccgcctaaaa ctctttaaaa
     8461 cttattaaaa ctcttaaaac ccgcctggcc tgtgcataac tgtctggcca gcgcacagcc
     8521 gaacagctgc aaaaagcgcc tacccttcgg tcgctgcgct ccctacgccc cgccgcttcg
     8581 cgtcggccta tcgcggccgc tggccgctca aaaatggctg gcctacggcc aggcaatcta
     8641 ccagggcgcg gacaagccgc gccgtcgcca ctcgaccgcc ggcgcccaca tcaaggctcc
     8701 gagtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     8761 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atggctaaaa
     8821 tgagaatatc accggaattg aaaaaactga tcgaaaaata ccgctgcgta aaagatacgg
     8881 aaggaatgtc tcctgctaag gtatataagc tggtgggaga aaatgaaaac ctatatttaa
     8941 aaatgacgga cagccggtat aaagggacca cctatgatgt ggaacgggaa aaggacatga
     9001 tgctatggct ggaaggaaag ctgcctgttc caaaggtcct gcactttgaa cggcatgatg
     9061 gctggagcaa tctgctcatg agtgaggccg atggcgtcct ttgctcggaa gagtatgaag
     9121 atgaacaaag ccctgaaaag attatcgagc tgtatgcgga gtgcatcagg ctctttcact
     9181 ccatcgacat atcggattgt ccctatacga atagcttaga cagccgctta gccgaattgg
     9241 attacttact gaataacgat ctggccgatg tggattgcga aaactgggaa gaggacactc
     9301 catttaaaga tccgcgcgag ctgtatgatt ttttaaagac ggaaaagccc gaagaggaac
     9361 ttgtcttttc ccacggcgac ctgggagaca gcaacatctt tgtgaaagat ggcaaagtaa
     9421 gtggctttat tgatcttggg agaagcggca gggcggacaa gtggtatgac attgccttct
     9481 gcgtccggtc gctcagggag gatatcgggg aagaacagta tgtcgagcta ttttttgact
     9541 tactggggat caagcctgat tgggagaaaa taaaatatta tattttactg gatgaattgt
     9601 tttagctgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     9661 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     9721 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     9781 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     9841 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     9901 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     9961 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
    10021 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
    10081 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
    10141 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
    10201 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
    10261 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
    10321 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
    10381 cctttttacg gttcctgctc ggatctgttg gaccggacag tagtcatggt tgatgggctg
    10441 cctgtatcga gtggtgattt tgtgccgagc tgccggtcgg ggagctgttg gctggctggt
    10501 ggcaggatat attgtggtgt aaacaaattg acgcttagac aacttaataa cacattgcgg
    10561 acgtttttaa tgtactgggg ttgaacactc t
//
