LOCUS       sequence-272863-       15673 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  STU2.0 SpyCas9 RZ system for plant genome editing.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15673)
  AUTHORS   Tang X, Ren Q, Yang L, Bao Y, Zhong Z, He Y, Liu S, Qi C, Liu B, 
            Wang Y, Sretenovic S, Zhang Y, Zheng X, Zhang T, Qi Y, Zhang Y
  TITLE     Single transcript unit CRISPR 2.0 systems for robust Cas9 and Cas12a
            mediated plant genome editing.
  JOURNAL   Plant Biotechnol J. 2019 Jul;17(7):1431-1445. doi: 
            10.1111/pbi.13068. Epub 2019 Jan 17.
  PUBMED    30582653
REFERENCE   2  (bases 1 to 15673)
  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"; doi: "10.1111/pbi.13068";
            journalName: "Plant Biotechnol J"; date: "2019-07"; volume: "17"; 
            issue: "7"; pages: "1431-1445"
COMMENT     Sequence Label: pGEL029
FEATURES             Location/Qualifiers
     source          1..15673
                     /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             complement(6250..6555)
                     /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(6352..6371)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     misc_RNA        6882..6957
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      7020..7268
                     /label=HSP terminator
                     /note="efficient transcription terminator from the 
                     Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya 
                     et al., 2010)"
     misc_feature    7289..7313
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             8614..9243
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             9672..10745
                     /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      10811..11005
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(11241..11263)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    11339..11479
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     11394..11413
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(11494..11511)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11665..12253)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11745..11764)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(12340..13134)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    13559..13583
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    13661..13835
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     promoter        complement(14925..15601)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     primer_bind     complement(14968..14988)
                     /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 gaaggtttga ggatctaaaa
     6181 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa awaramammr
     6241 rarraracct tatattcccc agaacatcag gttaatggcg tttttgatgt cattttcgcg
     6301 gtggctgaga tcagccactt cttccccgat aacggaaacc ggcacactgg ccatatcggt
     6361 ggtcatcatg cgccagcttt catccccgat atgcaccacc gggtaaagtt cacgggagac
     6421 tttatctgac agcagacgtg cactggccag ggggatcacc atccgtcgcc cgggcgtgtc
     6481 aataatatca ctctgtacat ccacaaacag acgataacgg ctctctcttt tataggtgta
     6541 aaccttaaac tgcatttcac cagcccctgt tctcgtcagc aaaagagccg ttcatttcaa
     6601 taaaccgggc gacctcagcc atcccttcct gattttccgc tttccagcgt tcggcacgca
     6661 gacgacgggc ttcattctgc atggttgtgc ttaccagacc ggagatattg acatcatata
     6721 tgccttgagc aactgatagc tgtcgctgtc aactgtcact gtaatacgct gcttcatagc
     6781 atacctcttt ttgacatact tcgggtatac atatcagtat atattcttat accgcaaaaa
     6841 tcagcgcgca aatacgcata ctgttatctg gcttggtctc agttttagag ctagaaatag
     6901 caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt
     6961 tgtcaccgga tgtgttttcc ggtctgatga gtccgtgagg acgaaacaaa agcgagctat
     7021 atgaagatga agatgaaata tttggtgtgt caaataaaaa gcttgtgtgc ttaagtttgt
     7081 gtttttttct tggcttgttg tgttatgaat ttgtggcttt ttctaatatt aaatgaatgt
     7141 aagatcacat tataatgaat aaacaaatgt ttctataatc cattgtgaat gttttgttgg
     7201 atctcttctg cagcatataa ctactgtatg tgctatggta tggactatgg aatatgatta
     7261 aagataagga gctcaaacta tcagtgtttg acaggatata ttggcgggta aacctaagag
     7321 aaaagagcgt ttattagaat aatcggatat ttaaaagggc gtgaaaaggt ttatccgttc
     7381 gtccatttgt atgtgcatgc caaccacagg gttcccctcg ggatcaaagt actttgatcc
     7441 aacccctccg ctgctatagt gcagtcggct tctgacgttc agtgcagccg tcttctgaaa
     7501 acgacatgtc gcacaagtcc taagttacgc gacaggctgc cgccctgccc ttttcctggc
     7561 gttttcttgt cgcgtgtttt agtcgcataa agtagaatac ttgcgactag aaccggagac
     7621 attacgccat gaacaagagc gccgccgctg gcctgctggg ctatgcccgc gtcagcaccg
     7681 acgaccagga cttgaccaac caacgggccg aactgcacgc ggccggctgc accaagctgt
     7741 tttccgagaa gatcaccggc accaggcgcg accgcccgga gctggccagg atgcttgacc
     7801 acctacgccc tggcgacgtt gtgacagtga ccaggctaga ccgcctggcc cgcagcaccc
     7861 gcgacctact ggacattgcc gagcgcatcc aggaggccgg cgcgggcctg cgtagcctgg
     7921 cagagccgtg ggccgacacc accacgccgg ccggccgcat ggtgttgacc gtgttcgccg
     7981 gcattgccga gttcgagcgt tccctaatca tcgaccgcac ccggagcggg cgcgaggccg
     8041 ccaaggcccg aggcgtgaag tttggccccc gccctaccct caccccggca cagatcgcgc
     8101 acgcccgcga gctgatcgac caggaaggcc gcaccgtgaa agaggcggct gcactgcttg
     8161 gcgtgcatcg ctcgaccctg taccgcgcac ttgagcgcag cgaggaagtg acgcccaccg
     8221 aggccaggcg gcgcggtgcc ttccgtgagg acgcattgac cgaggccgac gccctggcgg
     8281 ccgccgagaa tgaacgccaa gaggaacaag catgaaaccg caccaggacg gccaggacga
     8341 accgtttttc attaccgaag agatcgaggc ggagatgatc gcggccgggt acgtgttcga
     8401 gccgcccgcg cacggctcaa ccgtgcggct gcatgaaatc ctggccggtt tgtctgatgc
     8461 caagctggcg gcctggccgg ccagcttggc cgctgaagaa accgagcgcc gccgtctaaa
     8521 aaggtgatgt gtatttgagt aaaacagctt gcgtcatgcg gtcgctgcgt atatgatgcg
     8581 atgagtaaat aaacaaatac gcaaggggaa cgcatgaagg ttatcgctgt acttaaccag
     8641 aaaggcgggt caggcaagac gaccatcgca acccatctag cccgcgccct gcaactcgcc
     8701 ggggccgatg ttctgttagt cgattccgat ccccagggca gtgcccgcga ttgggcggcc
     8761 gtgcgggaag atcaaccgct aaccgttgtc ggcatcgacc gcccgacgat tgaccgcgac
     8821 gtgaaggcca tcggccggcg cgacttcgta gtgatcgacg gagcgcccca ggcggcggac
     8881 ttggctgtgt ccgcgatcaa ggcagccgac ttcgtgctga ttccggtgca gccaagccct
     8941 tacgacatat gggccaccgc cgacctggtg gagctggtta agcagcgcat tgaggtcacg
     9001 gatggaaggc tacaagcggc ctttgtcgtg tcgcgggcga tcaaaggcac gcgcatcggc
     9061 ggtgaggttg ccgaggcgct ggccgggtac gagctgccca ttcttgagtc ccgtatcacg
     9121 cagcgcgtga gctacccagg cactgccgcc gccggcacaa ccgttcttga atcagaaccc
     9181 gagggcgacg ctgcccgcga ggtccaggcg ctggccgctg aaattaaatc aaaactcatt
     9241 tgagttaatg aggtaaagag aaaatgagca aaagcacaaa cacgctaagt gccggccgtc
     9301 cgagcgcacg cagcagcaag gctgcaacgt tggccagcct ggcagacacg ccagccatga
     9361 agcgggtcaa ctttcagttg ccggcggagg atcacaccaa gctgaagatg tacgcggtac
     9421 gccaaggcaa gaccattacc gagctgctat ctgaatacat cgcgcagcta ccagagtaaa
     9481 tgagcaaatg aataaatgag tagatgaatt ttagcggcta aaggaggcgg catggaaaat
     9541 caagaacaac caggcaccga cgccgtggaa tgccccatgt gtggaggaac gggcggttgg
     9601 ccaggcgtaa gcggctgggt tgcctgccgg ccctgcaatg gcactggaac ccccaagccc
     9661 gaggaatcgg cgtgagcggt cgcaaaccat ccggcccggt acaaatcggc gcggcgctgg
     9721 gtgatgacct ggtggagaag ttgaaggccg cgcaggccgc ccagcggcaa cgcatcgagg
     9781 cagaagcacg ccccggtgaa tcgtggcaag cggccgctga tcgaatccgc aaagaatccc
     9841 ggcaaccgcc ggcagccggt gcgccgtcga ttaggaagcc gcccaagggc gacgagcaac
     9901 cagatttttt cgttccgatg ctctatgacg tgggcacccg cgatagtcgc agcatcatgg
     9961 acgtggccgt tttccgtctg tcgaagcgtg accgacgagc tggcgaggtg atccgctacg
    10021 agcttccaga cgggcacgta gaggtttccg cagggccggc cggcatggcc agtgtgtggg
    10081 attacgacct ggtactgatg gcggtttccc atctaaccga atccatgaac cgataccggg
    10141 aagggaaggg agacaagccc ggccgcgtgt tccgtccaca cgttgcggac gtactcaagt
    10201 tctgccggcg agccgatggc ggaaagcaga aagacgacct ggtagaaacc tgcattcggt
    10261 taaacaccac gcacgttgcc atgcagcgta cgaagaaggc caagaacggc cgcctggtga
    10321 cggtatccga gggtgaagcc ttgattagcc gctacaagat cgtaaagagc gaaaccgggc
    10381 ggccggagta catcgagatc gagctagctg attggatgta ccgcgagatc acagaaggca
    10441 agaacccgga cgtgctgacg gttcaccccg attacttttt gatcgatccc ggcatcggcc
    10501 gttttctcta ccgcctggca cgccgcgccg caggcaaggc agaagccaga tggttgttca
    10561 agacgatcta cgaacgcagt ggcagcgccg gagagttcaa gaagttctgt ttcaccgtgc
    10621 gcaagctgat cgggtcaaat gacctgccgg agtacgattt gaaggaggag gcggggcagg
    10681 ctggcccgat cctagtcatg cgctaccgca acctgatcga gggcgaagca tccgccggtt
    10741 cctaatgtac ggagcagatg ctagggcaaa ttgccctagc aggggaaaaa ggtcgaaaag
    10801 gcctctttcc tgtggatagc acgtacattg ggaacccaaa gccgtacatt gggaaccgga
    10861 acccgtacat tgggaaccca aagccgtaca ttgggaaccg gtcacacatg taagtgactg
    10921 atataaaaga gaaaaaaggc gatttttccg cctaaaactc tttaaaactt attaaaactc
    10981 ttaaaacccg cctggcctgt gcataactgt ctggccagcg cacagccgaa gagctgcaaa
    11041 aagcgcctac ccttcggtcg ctgcgctccc tacgccccgc cgcttcgcgt cggcctatcg
    11101 cggccgctgg ccgctcaaaa atggctggcc tacggccagg caatctacca gggcgcggac
    11161 aagccgcgcc gtcgccactc gaccgccggc gcccacatca aggcaccctg cctcgcgcgt
    11221 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggaaacggt cacagcttgt
    11281 ctgtaagcgg atgccgggag cagacaagcg cgtcagcggg tgttggcggg tgtcggggcg
    11341 cagccatgac ccagtcacgt agcgatagcg gagtgtatac tggcttaact atgcggcatc
    11401 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
    11461 gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
    11521 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
    11581 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
    11641 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
    11701 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
    11761 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
    11821 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
    11881 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
    11941 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
    12001 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
    12061 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
    12121 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
    12181 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
    12241 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
    12301 aaactcacgt taagggattt tggtcatgca ttctaggtac taaaacaatt catccagtaa
    12361 aatataatat tttattttct cccaatcagg cttgatcccc agtaagtcaa aaaatagctc
    12421 gacatactgt tcttccccga tatcctccct gatcgaccgg acgcagaagg caatgtcata
    12481 ccacttgtcc gccctgccgc ttctcccaag atcaataaag ccacttactt tgccatcttt
    12541 cacaaagatg ttgctgtctc ccaggtcgcc gtgggaaaag acaagttcct cttcgggctt
    12601 ttccgtcttt aaaaaatcat acagctcgcg cggatcttta aatggagtgt cttcttccca
    12661 gttttcgcaa tccacatcgg ccagatcgtt attcagtaag taatccaatt cggctaagcg
    12721 gctgtctaag ctattcgtat agggacaatc cgatatgtcg atggagtgaa agagcctgat
    12781 gcactccgca tacagctcga taatcttttc agggctttgt tcatcttcat actcttccga
    12841 gcaaaggacg ccatcggcct cactcatgag cagattgctc cagccatcat gccgttcaaa
    12901 gtgcaggacc tttggaacag gcagctttcc ttccagccat agcatcatgt ccttttcccg
    12961 ttccacatca taggtggtcc ctttataccg gctgtccgtc atttttaaat ataggttttc
    13021 attttctccc accagcttat ataccttagc aggagacatt ccttccgtat cttttacgca
    13081 gcggtatttt tcgatcagtt ttttcaattc cggtgatatt ctcattttag ccatttatta
    13141 tttccttcct cttttctaca gtatttaaag ataccccaag aagctaatta taacaagacg
    13201 aactccaatt cactgttcct tgcattctaa aaccttaaat accagaaaac agctttttca
    13261 aagttgtttt caaagttggc gtataacata gtatcgacgg agccgatttt gaaaccgcgg
    13321 tgatcacagg cagcaacgct ctgtcatcgt tacaatcaac atgctaccct ccgcgagatc
    13381 atccgtgttt caaacccggc agcttagttg ccgttcttcc gaatagcatc ggtaacatga
    13441 gcaaagtctg ccgccttaca acggctctcc cgctgacgcc gtcccggact gatgggctgc
    13501 ctgtatcgag tggtgatttt gtgccgagct gccggtcggg gagctgttgg ctggctggtg
    13561 gcaggatata ttgtggtgta aacaaattga cgcttagaca acttaataac acattgcgga
    13621 cgtttttaat gtactgaatt aacgccgaat taattcgggg gatctggatt ttagtactgg
    13681 attttggttt taggaattag aaattttatt gatagaagta ttttacaaat acaaatacat
    13741 actaagggtt tcttatatgc tcaacacatg agcgaaaccc tataggaacc ctaattccct
    13801 tatctgggaa ctactcacac attattatgg agaaaataga gcttgtcgat cgttaattaa
    13861 tcacctccaa gatcccggac gagtgctggg gcgtcggttt ccactatcgg cgagtacttc
    13921 tacacagcca tcggtccaga cggccgcgct tctgcgggcg atttgtgtac gcccgacagt
    13981 cccggctccg gatcggacga ttgcgtcgca tcgaccctgc gcccaagctg catcatcgaa
    14041 attgccgtca accaagctct gatagagttg gtcaagacca atgcggagca tatacgcccg
    14101 gagccgcggc gatcctgcaa gctccggatg cctccgctcg aagtagcgcg tctgctgctc
    14161 catacaagcc aaccacggcc tccagaagaa gatgttggcg acctcgtatt gggaatcccc
    14221 gaacatcgcc tcgctccagt caatgaccgc tgttatgcgg ccattgtccg tcaggacatt
    14281 gttggagccg aaatccgcgt gcacgaggtg ccggacttcg gggcagtcct cggcccaaag
    14341 catcagctca tcgagagcct gcgcgacgga cgcactgacg gtgtcgtcca tcacagtttg
    14401 ccagtgatac acatggggat cagcaatcgc gcatatgaaa tcacgccatg tagtgtattg
    14461 accgattcct tgcggtccga atgggccgaa cccgctcgtc tggctaagat cggccgcagc
    14521 gatcgcatcc atggcctccg cgaccggctg cagaacagcg ggcagttcgg tttcaggcag
    14581 gtcttgcaac gtgacaccct gtgcacggcg ggagatgcaa taggtcaggc tctcgctgaa
    14641 ttccccaatg tcaagcactt ccggaatcgg gagcgcggcc gatgcaaagt gccgataaac
    14701 ataacgatct ttgtagaaac catcggcgca gctatttacc cgcaggacat atccacgccc
    14761 tcctacatcg aagctgaaag cacgagattc ttcgccctcc gagagctgca tcaggtcgga
    14821 cacgctgtcg aacttttcga tcagaaactt ctcgacagac gtcgcggtga gttcaggctt
    14881 tttcatccta ggagagatag atttgtagag agagactggt gatttcagcg tgtcctctcc
    14941 aaatgaaatg aacttcctta tatagaggaa gggtcttgcg aaggatagtg ggattgtgcg
    15001 tcatccctta cgtcagtgga gatatcacat caatccactt gctttgaaga cgtggttgga
    15061 acgtcttctt tttccacgat gctcctcgtg ggtgggggtc catctttggg accactgtcg
    15121 gcagaggcat cttgaacgat agcctttcct ttatcgcaat gatggcattt gtaggtgcca
    15181 ccttcctttt ctactgtcct tttgatgaag tgacagatag ctgggcaatg gaatccgagg
    15241 aggtttcccg atattaccct ttgttgaaaa gtctcaatag ccctttggtc ttctgagact
    15301 gtatctttga tattcttgga gtagacgaga gtgtcgtgct ccaccatgtt cacatcaatc
    15361 cacttgcttt gaagacgtgg ttggaacgtc ttctttttcc acgatgctcc tcgtgggtgg
    15421 gggtccatct ttgggaccac tgtcggcaga ggcatcttga acgatagcct ttcctttatc
    15481 gcaatgatgg catttgtagg tgccaccttc cttttctact gtccttttga tgaagtgaca
    15541 gatagctggg caatggaatc cgaggaggtt tcccgatatt accctttgtt gaaaagtctc
    15601 aatagccctt tggtcttctg agactgtatc tttgatattc ttggagtaga cgagagtgtc
    15661 gtgctccacc atg
//
