LOCUS       sequence-255725-       15911 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For genome editing in plant by SpCas9 wild type.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15911)
  AUTHORS   Zhong Z, Sretenovic S, Ren Q, Yang L, Bao Y, Qi C, Yuan M, He Y, Liu
            S, Liu X, Wang J, Huang L, Wang Y, Baby D, Wang D, Zhang T, Qi Y, 
            Zhang Y
  TITLE     Improving Plant Genome Editing with High-Fidelity xCas9 and 
            Non-canonical PAM-Targeting Cas9-NG.
  JOURNAL   Mol Plant. 2019 Mar 27. pii: S1674-2052(19)30124-8. doi: 
            10.1016/j.molp.2019.03.011.
  PUBMED    30928637
REFERENCE   2  (bases 1 to 15911)
  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: "Mol Plant. 
            2019 Mar 27. pii: S1674-2052(19)30124-8. doi: 
            10.1016/j.molp.2019.03.011."
COMMENT     Sequence Label: pGEL062
FEATURES             Location/Qualifiers
     source          1..15911
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        8..2006
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
     CDS             2031..6134
                     /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             2031..6134
                     /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             6147..6167
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6180..6200
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6213..6233
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      6244..6492
                     /label=HSP terminator
                     /note="efficient transcription terminator from the 
                     Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya 
                     et al., 2010)"
     CDS             complement(6754..7059)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(6856..6875)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     misc_RNA        7386..7461
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(7473..7497)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    7527..7551
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             8852..9481
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             9910..10983
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      11049..11243
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(11479..11501)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    11577..11717
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     11632..11651
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(11732..11749)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11903..12491)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11983..12002)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(12578..13372)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    13797..13821
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    13899..14073
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     promoter        complement(15163..15839)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     primer_bind     complement(15206..15226)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
ORIGIN
        1 ggcgcgcctg cagtgcagcg tgacccggtc gtgcccctct cttgagataa ggtgagcatt
       61 gcatgtctaa gttataaaaa attaccacat attttttttg tcacacttgt ttgaagtgca
      121 gtttatctat ctttatacat atatttaaac tttactctac gaataatata atctatagta
      181 ctacaataat atcagtgttt tagagaatca tataaatgaa cagttagaca tggtctaaag
      241 gacaattgag tattttgaca acaggactct acagttttat ctttttagtg tgcatgtgtt
      301 ctcctttttt tttgcaaata gcttcaccta tataatactt catccatttt attagtacat
      361 ccatttaggg tttagggtta atggttttta tagactaatt tttttagtac atctatttta
      421 ttctatttta gcctctaaat taagaaaact aaaactctat tttagttttt ttatttaata
      481 atttagatat aaaatagaat aaaataaagt gactaaaaat taaacaaata ccctttaaga
      541 aattaaaaaa actaaggaaa catttttctt gtttcgagta gataatgcca gcctgttaaa
      601 cgccgtcgac gagtctaacg gacaccaacc agcgaaccag cagcgtcgcg tcgggccaag
      661 cgaagcagac ggcacggcat ctctgtcgct gcctctggac ccctctcgag agttccgctc
      721 caccgttgga cttgctccgc tgtcggcatc cagaaattgc gtggcggagc ggcagacgtg
      781 agccggcacg gcaggcggcc tcctcctcct ctcacggcac cggcagctac gggggattcc
      841 tttcccaccg ctccttcgct ttcccttcct cgcccgccgt aataaataga caccccctcc
      901 acaccctctt tccccaacct cgtgttgttc ggagcgcaca cacacacaac cagaactccc
      961 ccaaatccac ccgtcggcac ctccgcttca aggtacgccg ctcgtcctcc cccccccccc
     1021 ctctctacct tctcaagatc ggcgttccgg tccatggtta gggcccggta gttctacttc
     1081 tgttcatgtt tgtgttagat ccgtgtttgt gttagatccg tgctactagc gttcgtacac
     1141 ggatgcgacc tgtacgtcag acacgttctg attgctaact tgccagtgtt tctctttggg
     1201 gaatcctggg atggctctag ccgttccgca gacgggatcg atttcatgat tttttttgtt
     1261 tcgttgcata gggtttggtt tgcccttttc ctttatttca atatatgccg tgcacttgtt
     1321 tgtcgggtca tcttttcatg cttttttttg tcttggttgt gatgatgtgg tctggttggg
     1381 cggtcgttca agatcggagt agaattaatt ctgtttcaaa ctacctggtg gatttattaa
     1441 ttttggatct gtatgtgtgt gccatacata ttcatagtta cgaattgaag atgatggatg
     1501 gaaatatcga tctaggatag gtatacatgt tgatgcgggt tttactgatg catatacaga
     1561 gatgcttttt gttcgcttgg ttgtgatgat gtggtgtggt tgggcggtcg ttcattcgtt
     1621 caagatcgga gtagaatact gtttcaaact acctggtgta tttattaatt ttggaactgt
     1681 atgtgtgtgt catacatctt catagttacg agtttaagat ggatggaaat atcgatctag
     1741 gataggtata catgttgatg tgggttttac tgatgcatat acatgatggc atatgcagca
     1801 tctattcata tgctctaacc ttgagtacct atctattata ataaacaagt atgttttata
     1861 attattttga tcttgatata cttggatgat ggcatatgca gcagctatat gtggattttt
     1921 ttagccctgc cttcatacgc tatttatttg cttggtactg tttcttttgt cgatgctcac
     1981 cctgttgttt ggtgttactt ctgcagcctg caggtagatc gctcgtcgac atggataaga
     2041 agtactctat cggactcgat atcggaacta actctgtggg atgggctgtg atcaccgatg
     2101 agtacaaggt gccatctaag aagttcaagg ttctcggaaa caccgatagg cactctatca
     2161 agaaaaacct tatcggtgct ctcctcttcg attctggtga aactgctgag gctaccagac
     2221 tcaagagaac cgctagaaga aggtacacca gaagaaagaa caggatctgc tacctccaag
     2281 agatcttctc taacgagatg gctaaagtgg atgattcatt cttccacagg ctcgaagagt
     2341 cattcctcgt ggaagaagat aagaagcacg agaggcaccc tatcttcgga aacatcgttg
     2401 atgaggtggc ataccacgag aagtacccta ctatctacca cctcagaaag aagctcgttg
     2461 attctactga taaggctgat ctcaggctca tctacctcgc tctcgctcac atgatcaagt
     2521 tcagaggaca cttcctcatc gagggtgatc tcaaccctga taactctgat gtggataagt
     2581 tgttcatcca gctcgtgcag acctacaacc agcttttcga agagaaccct atcaacgctt
     2641 caggtgtgga tgctaaggct atcctctctg ctaggctctc taagtcaaga aggcttgaga
     2701 acctcattgc tcagctccct ggtgagaaga agaacggact tttcggaaac ttgatcgctc
     2761 tctctctcgg actcacccct aacttcaagt ctaacttcga tctcgctgag gatgcaaagc
     2821 tccagctctc aaaggatacc tacgatgatg atctcgataa cctcctcgct cagatcggag
     2881 atcagtacgc tgatttgttc ctcgctgcta agaacctctc tgatgctatc ctcctcagtg
     2941 atatcctcag agtgaacacc gagatcacca aggctccact ctcagcttct atgatcaaga
     3001 gatacgatga gcaccaccag gatctcacac ttctcaaggc tcttgttaga cagcagctcc
     3061 cagagaagta caaagagatt ttcttcgatc agtctaagaa cggatacgct ggttacatcg
     3121 atggtggtgc atctcaagaa gagttctaca agttcatcaa gcctatcctc gagaagatgg
     3181 atggaaccga ggaactcctc gtgaagctca atagagagga tcttctcaga aagcagagga
     3241 ccttcgataa cggatctatc cctcatcaga tccacctcgg agagttgcac gctatcctta
     3301 gaaggcaaga ggatttctac ccattcctca aggataacag ggaaaagatt gagaagattc
     3361 tcaccttcag aatcccttac tacgtgggac ctctcgctag aggaaactca agattcgctt
     3421 ggatgaccag aaagtctgag gaaaccatca ccccttggaa cttcgaagag gtggtggata
     3481 agggtgctag tgctcagtct ttcatcgaga ggatgaccaa cttcgataag aaccttccaa
     3541 acgagaaggt gctccctaag cactctttgc tctacgagta cttcaccgtg tacaacgagt
     3601 tgaccaaggt taagtacgtg accgagggaa tgaggaagcc tgcttttttg tcaggtgagc
     3661 aaaagaaggc tatcgttgat ctcttgttca agaccaacag aaaggtgacc gtgaagcagc
     3721 tcaaagagga ttacttcaag aaaatcgagt gcttcgattc agttgagatt tctggtgttg
     3781 aggataggtt caacgcatct ctcggaacct accacgatct cctcaagatc attaaggata
     3841 aggatttctt ggataacgag gaaaacgagg atatcttgga ggatatcgtt cttaccctca
     3901 ccctctttga agatagagag atgattgaag aaaggctcaa gacctacgct catctcttcg
     3961 atgataaggt gatgaagcag ttgaagagaa gaagatacac tggttgggga aggctctcaa
     4021 gaaagctcat taacggaatc agggataagc agtctggaaa gacaatcctt gatttcctca
     4081 agtctgatgg attcgctaac agaaacttca tgcagctcat ccacgatgat tctctcacct
     4141 ttaaagagga tatccagaag gctcaggttt caggacaggg tgatagtctc catgagcata
     4201 tcgctaacct cgctggatct cctgcaatca agaagggaat cctccagact gtgaaggttg
     4261 tggatgagtt ggtgaaggtg atgggaaggc ataagcctga gaacatcgtg atcgaaatgg
     4321 ctagagagaa ccagaccact cagaagggac agaagaactc tagggaaagg atgaagagga
     4381 tcgaggaagg tatcaaagag cttggatctc agatcctcaa agagcaccct gttgagaaca
     4441 ctcagctcca gaatgagaag ctctacctct actacctcca gaacggaagg gatatgtatg
     4501 tggatcaaga gttggatatc aacaggctct ctgattacga tgttgatcat atcgtgccac
     4561 agtcattctt gaaggatgat tctatcgata acaaggtgct caccaggtct gataagaaca
     4621 ggggtaagag tgataacgtg ccaagtgaag aggttgtgaa gaaaatgaag aactattgga
     4681 ggcagctcct caacgctaag ctcatcactc agagaaagtt cgataacttg actaaggctg
     4741 agaggggagg actctctgaa ttggataagg caggattcat caagaggcag cttgtggaaa
     4801 ccaggcagat cactaagcac gttgcacaga tcctcgattc taggatgaac accaagtacg
     4861 atgagaacga taagttgatc agggaagtga aggttatcac cctcaagtca aagctcgtgt
     4921 ctgatttcag aaaggatttc caattctaca aggtgaggga aatcaacaac taccaccacg
     4981 ctcacgatgc ttaccttaac gctgttgttg gaaccgctct catcaagaag tatcctaagc
     5041 tcgagtcaga gttcgtgtac ggtgattaca aggtgtacga tgtgaggaag atgatcgcta
     5101 agtctgagca agagatcgga aaggctaccg ctaagtattt cttctactct aacatcatga
     5161 atttcttcaa gaccgagatt accctcgcta acggtgagat cagaaagagg ccactcatcg
     5221 agacaaacgg tgaaacaggt gagatcgtgt gggataaggg aagggatttc gctaccgtta
     5281 gaaaggtgct ctctatgcca caggtgaaca tcgttaagaa aaccgaggtg cagaccggtg
     5341 gattctctaa agagtctatc ctccctaaga ggaactctga taagctcatt gctaggaaga
     5401 aggattggga ccctaagaaa tacggtggtt tcgattctcc taccgtggct tactctgttc
     5461 tcgttgtggc taaggttgag aagggaaaga gtaagaagct caagtctgtt aaggaacttc
     5521 tcggaatcac tatcatggaa aggtcatctt tcgagaagaa cccaatcgat ttcctcgagg
     5581 ctaagggata caaagaggtt aagaaggatc tcatcatcaa gctcccaaag tactcactct
     5641 tcgaactcga gaacggtaga aagaggatgc tcgcttctgc tggtgagctt caaaagggaa
     5701 acgagcttgc tctcccatct aagtacgtta actttcttta cctcgcttct cactacgaga
     5761 agttgaaggg atctccagaa gataacgagc agaagcaact tttcgttgag cagcacaagc
     5821 actacttgga tgagatcatc gagcagatct ctgagttctc taaaagggtg atcctcgctg
     5881 atgcaaacct cgataaggtg ttgtctgctt acaacaagca cagagataag cctatcaggg
     5941 aacaggcaga gaacatcatc catctcttca cccttaccaa cctcggtgct cctgctgctt
     6001 tcaagtactt cgatacaacc atcgatagga agagatacac ctctaccaaa gaagtgctcg
     6061 atgctaccct catccatcag tctatcactg gactctacga gactaggatc gatctctcac
     6121 agctcggtgg tgattcaagg gctgatccta agaagaagag gaaggtttct agggcagatc
     6181 caaagaagaa gcggaaggtg tctagagctg atccgaagaa gaagcgcaag gtgtgactcg
     6241 agatatgaag atgaagatga aatatttggt gtgtcaaata aaaagcttgt gtgcttaagt
     6301 ttgtgttttt ttcttggctt gttgtgttat gaatttgtgg ctttttctaa tattaaatga
     6361 atgtaagatc acattataat gaataaacaa atgtttctat aatccattgt gaatgttttg
     6421 ttggatctct tctgcagcat ataactactg tatgtgctat ggtatggact atggaatatg
     6481 attaaagata agaggatccg cggatcatga accaacggcc tggctgtatt tggtggttgt
     6541 gtagggagat ggggagaaga aaagcccgat tctcttcgct gtgatgggct ggatgcatgc
     6601 gggggagcgg gaggcccaag tacgtgcacg gtgagcggcc cacagggcga gtgtgagcgc
     6661 gagaggcggg aggaacagtt tagtaccaca ttgcccagct aactcgaacg cgaccaactt
     6721 ataaacccgc gcgctgtcgc ttgtgtggag accttatatt ccccagaaca tcaggttaat
     6781 ggcgtttttg atgtcatttt cgcggtggct gagatcagcc acttcttccc cgataacgga
     6841 aaccggcaca ctggccatat cggtggtcat catgcgccag ctttcatccc cgatatgcac
     6901 caccgggtaa agttcacggg agactttatc tgacagcaga cgtgcactgg ccagggggat
     6961 caccatccgt cgcccgggcg tgtcaataat atcactctgt acatccacaa acagacgata
     7021 acggctctct cttttatagg tgtaaacctt aaactgcatt tcaccagccc ctgttctcgt
     7081 cagcaaaaga gccgttcatt tcaataaacc gggcgacctc agccatccct tcctgatttt
     7141 ccgctttcca gcgttcggca cgcagacgac gggcttcatt ctgcatggtt gtgcttacca
     7201 gaccggagat attgacatca tatatgcctt gagcaactga tagctgtcgc tgtcaactgt
     7261 cactgtaata cgctgcttca tagcatacct ctttttgaca tacttcgggt atacatatca
     7321 gtatatattc ttataccgca aaaatcagcg cgcaaatacg catactgtta tctggcttgg
     7381 tctcagtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa
     7441 aaagtggcac cgagtcggtg ctttttttct agacccagct ttcttgtaca aagttggcat
     7501 tactgcgagc tcaaactatc agtgtttgac aggatatatt ggcgggtaaa cctaagagaa
     7561 aagagcgttt attagaataa tcggatattt aaaagggcgt gaaaaggttt atccgttcgt
     7621 ccatttgtat gtgcatgcca accacagggt tcccctcggg atcaaagtac tttgatccaa
     7681 cccctccgct gctatagtgc agtcggcttc tgacgttcag tgcagccgtc ttctgaaaac
     7741 gacatgtcgc acaagtccta agttacgcga caggctgccg ccctgccctt ttcctggcgt
     7801 tttcttgtcg cgtgttttag tcgcataaag tagaatactt gcgactagaa ccggagacat
     7861 tacgccatga acaagagcgc cgccgctggc ctgctgggct atgcccgcgt cagcaccgac
     7921 gaccaggact tgaccaacca acgggccgaa ctgcacgcgg ccggctgcac caagctgttt
     7981 tccgagaaga tcaccggcac caggcgcgac cgcccggagc tggccaggat gcttgaccac
     8041 ctacgccctg gcgacgttgt gacagtgacc aggctagacc gcctggcccg cagcacccgc
     8101 gacctactgg acattgccga gcgcatccag gaggccggcg cgggcctgcg tagcctggca
     8161 gagccgtggg ccgacaccac cacgccggcc ggccgcatgg tgttgaccgt gttcgccggc
     8221 attgccgagt tcgagcgttc cctaatcatc gaccgcaccc ggagcgggcg cgaggccgcc
     8281 aaggcccgag gcgtgaagtt tggcccccgc cctaccctca ccccggcaca gatcgcgcac
     8341 gcccgcgagc tgatcgacca ggaaggccgc accgtgaaag aggcggctgc actgcttggc
     8401 gtgcatcgct cgaccctgta ccgcgcactt gagcgcagcg aggaagtgac gcccaccgag
     8461 gccaggcggc gcggtgcctt ccgtgaggac gcattgaccg aggccgacgc cctggcggcc
     8521 gccgagaatg aacgccaaga ggaacaagca tgaaaccgca ccaggacggc caggacgaac
     8581 cgtttttcat taccgaagag atcgaggcgg agatgatcgc ggccgggtac gtgttcgagc
     8641 cgcccgcgca cggctcaacc gtgcggctgc atgaaatcct ggccggtttg tctgatgcca
     8701 agctggcggc ctggccggcc agcttggccg ctgaagaaac cgagcgccgc cgtctaaaaa
     8761 ggtgatgtgt atttgagtaa aacagcttgc gtcatgcggt cgctgcgtat atgatgcgat
     8821 gagtaaataa acaaatacgc aaggggaacg catgaaggtt atcgctgtac ttaaccagaa
     8881 aggcgggtca ggcaagacga ccatcgcaac ccatctagcc cgcgccctgc aactcgccgg
     8941 ggccgatgtt ctgttagtcg attccgatcc ccagggcagt gcccgcgatt gggcggccgt
     9001 gcgggaagat caaccgctaa ccgttgtcgg catcgaccgc ccgacgattg accgcgacgt
     9061 gaaggccatc ggccggcgcg acttcgtagt gatcgacgga gcgccccagg cggcggactt
     9121 ggctgtgtcc gcgatcaagg cagccgactt cgtgctgatt ccggtgcagc caagccctta
     9181 cgacatatgg gccaccgccg acctggtgga gctggttaag cagcgcattg aggtcacgga
     9241 tggaaggcta caagcggcct ttgtcgtgtc gcgggcgatc aaaggcacgc gcatcggcgg
     9301 tgaggttgcc gaggcgctgg ccgggtacga gctgcccatt cttgagtccc gtatcacgca
     9361 gcgcgtgagc tacccaggca ctgccgccgc cggcacaacc gttcttgaat cagaacccga
     9421 gggcgacgct gcccgcgagg tccaggcgct ggccgctgaa attaaatcaa aactcatttg
     9481 agttaatgag gtaaagagaa aatgagcaaa agcacaaaca cgctaagtgc cggccgtccg
     9541 agcgcacgca gcagcaaggc tgcaacgttg gccagcctgg cagacacgcc agccatgaag
     9601 cgggtcaact ttcagttgcc ggcggaggat cacaccaagc tgaagatgta cgcggtacgc
     9661 caaggcaaga ccattaccga gctgctatct gaatacatcg cgcagctacc agagtaaatg
     9721 agcaaatgaa taaatgagta gatgaatttt agcggctaaa ggaggcggca tggaaaatca
     9781 agaacaacca ggcaccgacg ccgtggaatg ccccatgtgt ggaggaacgg gcggttggcc
     9841 aggcgtaagc ggctgggttg cctgccggcc ctgcaatggc actggaaccc ccaagcccga
     9901 ggaatcggcg tgagcggtcg caaaccatcc ggcccggtac aaatcggcgc ggcgctgggt
     9961 gatgacctgg tggagaagtt gaaggccgcg caggccgccc agcggcaacg catcgaggca
    10021 gaagcacgcc ccggtgaatc gtggcaagcg gccgctgatc gaatccgcaa agaatcccgg
    10081 caaccgccgg cagccggtgc gccgtcgatt aggaagccgc ccaagggcga cgagcaacca
    10141 gattttttcg ttccgatgct ctatgacgtg ggcacccgcg atagtcgcag catcatggac
    10201 gtggccgttt tccgtctgtc gaagcgtgac cgacgagctg gcgaggtgat ccgctacgag
    10261 cttccagacg ggcacgtaga ggtttccgca gggccggccg gcatggccag tgtgtgggat
    10321 tacgacctgg tactgatggc ggtttcccat ctaaccgaat ccatgaaccg ataccgggaa
    10381 gggaagggag acaagcccgg ccgcgtgttc cgtccacacg ttgcggacgt actcaagttc
    10441 tgccggcgag ccgatggcgg aaagcagaaa gacgacctgg tagaaacctg cattcggtta
    10501 aacaccacgc acgttgccat gcagcgtacg aagaaggcca agaacggccg cctggtgacg
    10561 gtatccgagg gtgaagcctt gattagccgc tacaagatcg taaagagcga aaccgggcgg
    10621 ccggagtaca tcgagatcga gctagctgat tggatgtacc gcgagatcac agaaggcaag
    10681 aacccggacg tgctgacggt tcaccccgat tactttttga tcgatcccgg catcggccgt
    10741 tttctctacc gcctggcacg ccgcgccgca ggcaaggcag aagccagatg gttgttcaag
    10801 acgatctacg aacgcagtgg cagcgccgga gagttcaaga agttctgttt caccgtgcgc
    10861 aagctgatcg ggtcaaatga cctgccggag tacgatttga aggaggaggc ggggcaggct
    10921 ggcccgatcc tagtcatgcg ctaccgcaac ctgatcgagg gcgaagcatc cgccggttcc
    10981 taatgtacgg agcagatgct agggcaaatt gccctagcag gggaaaaagg tcgaaaaggc
    11041 ctctttcctg tggatagcac gtacattggg aacccaaagc cgtacattgg gaaccggaac
    11101 ccgtacattg ggaacccaaa gccgtacatt gggaaccggt cacacatgta agtgactgat
    11161 ataaaagaga aaaaaggcga tttttccgcc taaaactctt taaaacttat taaaactctt
    11221 aaaacccgcc tggcctgtgc ataactgtct ggccagcgca cagccgaaga gctgcaaaaa
    11281 gcgcctaccc ttcggtcgct gcgctcccta cgccccgccg cttcgcgtcg gcctatcgcg
    11341 gccgctggcc gctcaaaaat ggctggccta cggccaggca atctaccagg gcgcggacaa
    11401 gccgcgccgt cgccactcga ccgccggcgc ccacatcaag gcaccctgcc tcgcgcgttt
    11461 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gaaacggtca cagcttgtct
    11521 gtaagcggat gccgggagca gacaagcgcg tcagcgggtg ttggcgggtg tcggggcgca
    11581 gccatgaccc agtcacgtag cgatagcgga gtgtatactg gcttaactat gcggcatcag
    11641 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
    11701 gaaaataccg catcaggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
    11761 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
    11821 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
    11881 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
    11941 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
    12001 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
    12061 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
    12121 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
    12181 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
    12241 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
    12301 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
    12361 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
    12421 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
    12481 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
    12541 actcacgtta agggattttg gtcatgcatt ctaggtacta aaacaattca tccagtaaaa
    12601 tataatattt tattttctcc caatcaggct tgatccccag taagtcaaaa aatagctcga
    12661 catactgttc ttccccgata tcctccctga tcgaccggac gcagaaggca atgtcatacc
    12721 acttgtccgc cctgccgctt ctcccaagat caataaagcc acttactttg ccatctttca
    12781 caaagatgtt gctgtctccc aggtcgccgt gggaaaagac aagttcctct tcgggctttt
    12841 ccgtctttaa aaaatcatac agctcgcgcg gatctttaaa tggagtgtct tcttcccagt
    12901 tttcgcaatc cacatcggcc agatcgttat tcagtaagta atccaattcg gctaagcggc
    12961 tgtctaagct attcgtatag ggacaatccg atatgtcgat ggagtgaaag agcctgatgc
    13021 actccgcata cagctcgata atcttttcag ggctttgttc atcttcatac tcttccgagc
    13081 aaaggacgcc atcggcctca ctcatgagca gattgctcca gccatcatgc cgttcaaagt
    13141 gcaggacctt tggaacaggc agctttcctt ccagccatag catcatgtcc ttttcccgtt
    13201 ccacatcata ggtggtccct ttataccggc tgtccgtcat ttttaaatat aggttttcat
    13261 tttctcccac cagcttatat accttagcag gagacattcc ttccgtatct tttacgcagc
    13321 ggtatttttc gatcagtttt ttcaattccg gtgatattct cattttagcc atttattatt
    13381 tccttcctct tttctacagt atttaaagat accccaagaa gctaattata acaagacgaa
    13441 ctccaattca ctgttccttg cattctaaaa ccttaaatac cagaaaacag ctttttcaaa
    13501 gttgttttca aagttggcgt ataacatagt atcgacggag ccgattttga aaccgcggtg
    13561 atcacaggca gcaacgctct gtcatcgtta caatcaacat gctaccctcc gcgagatcat
    13621 ccgtgtttca aacccggcag cttagttgcc gttcttccga atagcatcgg taacatgagc
    13681 aaagtctgcc gccttacaac ggctctcccg ctgacgccgt cccggactga tgggctgcct
    13741 gtatcgagtg gtgattttgt gccgagctgc cggtcgggga gctgttggct ggctggtggc
    13801 aggatatatt gtggtgtaaa caaattgacg cttagacaac ttaataacac attgcggacg
    13861 tttttaatgt actgaattaa cgccgaatta attcggggga tctggatttt agtactggat
    13921 tttggtttta ggaattagaa attttattga tagaagtatt ttacaaatac aaatacatac
    13981 taagggtttc ttatatgctc aacacatgag cgaaacccta taggaaccct aattccctta
    14041 tctgggaact actcacacat tattatggag aaaatagagc ttgtcgatcg ttaattaatc
    14101 acctccaaga tcccggacga gtgctggggc gtcggtttcc actatcggcg agtacttcta
    14161 cacagccatc ggtccagacg gccgcgcttc tgcgggcgat ttgtgtacgc ccgacagtcc
    14221 cggctccgga tcggacgatt gcgtcgcatc gaccctgcgc ccaagctgca tcatcgaaat
    14281 tgccgtcaac caagctctga tagagttggt caagaccaat gcggagcata tacgcccgga
    14341 gccgcggcga tcctgcaagc tccggatgcc tccgctcgaa gtagcgcgtc tgctgctcca
    14401 tacaagccaa ccacggcctc cagaagaaga tgttggcgac ctcgtattgg gaatccccga
    14461 acatcgcctc gctccagtca atgaccgctg ttatgcggcc attgtccgtc aggacattgt
    14521 tggagccgaa atccgcgtgc acgaggtgcc ggacttcggg gcagtcctcg gcccaaagca
    14581 tcagctcatc gagagcctgc gcgacggacg cactgacggt gtcgtccatc acagtttgcc
    14641 agtgatacac atggggatca gcaatcgcgc atatgaaatc acgccatgta gtgtattgac
    14701 cgattccttg cggtccgaat gggccgaacc cgctcgtctg gctaagatcg gccgcagcga
    14761 tcgcatccat ggcctccgcg accggctgca gaacagcggg cagttcggtt tcaggcaggt
    14821 cttgcaacgt gacaccctgt gcacggcggg agatgcaata ggtcaggctc tcgctgaatt
    14881 ccccaatgtc aagcacttcc ggaatcggga gcgcggccga tgcaaagtgc cgataaacat
    14941 aacgatcttt gtagaaacca tcggcgcagc tatttacccg caggacatat ccacgccctc
    15001 ctacatcgaa gctgaaagca cgagattctt cgccctccga gagctgcatc aggtcggaca
    15061 cgctgtcgaa cttttcgatc agaaacttct cgacagacgt cgcggtgagt tcaggctttt
    15121 tcatcctagg agagatagat ttgtagagag agactggtga tttcagcgtg tcctctccaa
    15181 atgaaatgaa cttccttata tagaggaagg gtcttgcgaa ggatagtggg attgtgcgtc
    15241 atcccttacg tcagtggaga tatcacatca atccacttgc tttgaagacg tggttggaac
    15301 gtcttctttt tccacgatgc tcctcgtggg tgggggtcca tctttgggac cactgtcggc
    15361 agaggcatct tgaacgatag cctttccttt atcgcaatga tggcatttgt aggtgccacc
    15421 ttccttttct actgtccttt tgatgaagtg acagatagct gggcaatgga atccgaggag
    15481 gtttcccgat attacccttt gttgaaaagt ctcaatagcc ctttggtctt ctgagactgt
    15541 atctttgata ttcttggagt agacgagagt gtcgtgctcc accatgttca catcaatcca
    15601 cttgctttga agacgtggtt ggaacgtctt ctttttccac gatgctcctc gtgggtgggg
    15661 gtccatcttt gggaccactg tcggcagagg catcttgaac gatagccttt cctttatcgc
    15721 aatgatggca tttgtaggtg ccaccttcct tttctactgt ccttttgatg aagtgacaga
    15781 tagctgggca atggaatccg aggaggtttc ccgatattac cctttgttga aaagtctcaa
    15841 tagccctttg gtcttctgag actgtatctt tgatattctt ggagtagacg agagtgtcgt
    15901 gctccaccat g
//
