LOCUS       sequence-182762-       13776 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  2x35S promoting Glycing Max codon optimized Cas9 with hygromycin 
            plant selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 13776)
  AUTHORS   Michno JM, Wang X, Liu J, Curtin SJ, Kono TJ, Stupar RM
  TITLE     CRISPR/Cas mutagenesis of soybean and Medicago truncatula using a 
            new web-tool and a modified Cas9 enzyme.
  JOURNAL   GM Crops Food. 2015;6(4):243-52. doi: 10.1080/21645698.2015.1106063.
  PUBMED    26479970
REFERENCE   2  (bases 1 to 13776)
  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: "GM Crops 
            Food."; date: "2015"; volume: "6(4):243-52. doi"; pages: " 
            10.1080/21645698.2015.1106063"
COMMENT     Sequence Label: Cas9 MDC32
FEATURES             Location/Qualifiers
     source          1..13776
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        411..755
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             816..4919
                     /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             816..4919
                     /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"
     RBS             4832..4840
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             4926..4946
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(5277..5293)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5277..5293)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5290..5312)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5301..5317
                     /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)."
     promoter        complement(join(5325..5331,5332..5349,5350..5355))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5370..5391
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5582..6258
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     CDS             6325..7350
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     polyA_signal    7391..7565
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    7643..7667
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             8092..8886
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      8973..9561
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9462..9481
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9715..9732
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    9747..9887
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(9813..9832)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9973..9995
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      10231..10425
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(10491..11564)
                     /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"
     CDS             complement(11993..12622)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
ORIGIN
        1 ctgcaggtca acatggtgga gcacgacaca cttgtctact ccaaaaatat caaagataca
       61 gtctcagaag accaaagggc aattgagact tttcaacaaa gggtaatatc cggaaacctc
      121 ctcggattcc attgcccagc tatctgtcac tttattgtga agatagtgga aaaggaaggt
      181 ggctcctaca aatgccatca ttgcgataaa ggaaaggcca tcgttgaaga tgcctctgcc
      241 gacagtggtc ccaaagatgg acccccaccc acgaggagca tcgtggaaaa agaagacgtt
      301 ccaaccacgt cttcaaagca agtggattga tgtgataaca tggtggagca cgacacactt
      361 gtctactcca aaaatatcaa agatacagtc tcagaagacc aaagggcaat tgagactttt
      421 caacaaaggg taatatccgg aaacctcctc ggattccatt gcccagctat ctgtcacttt
      481 attgtgaaga tagtggaaaa ggaaggtggc tcctacaaat gccatcattg cgataaagga
      541 aaggccatcg ttgaagatgc ctctgccgac agtggtccca aagatggacc cccacccacg
      601 aggagcatcg tggaaaaaga agacgttcca accacgtctt caaagcaagt ggattgatgt
      661 gatatctcca ctgacgtaag ggatgacgca caatcccact atccttcgca agacccttcc
      721 tctatataag gaagttcatt tcatttggag aggacctcga ctctagagga tccccgggta
      781 ccgggccccc cctcgaggcg cgccaccagg tcgacatgga taagaagtac tccatcggac
      841 tggatatcgg aactaactcc gtgggatggg ctgtgatcac tgatgagtac aaggtgccat
      901 ccaagaagtt caaggtgctg ggaaacactg atagacactc catcaagaag aacctgatcg
      961 gagctctgct gttcgattcc ggagagactg ctgaggctac tagactgaag agaactgcta
     1021 gaagaagata cactagaaga aagaacagaa tctgctacct gcaagagatc ttctccaacg
     1081 agatggctaa ggtggatgat tccttcttcc acagactgga ggagtccttc ctggtggagg
     1141 aggataagaa gcacgagaga cacccaatct tcggaaacat cgtggatgag gtggcttacc
     1201 acgagaagta cccaactatc taccacctga gaaagaagct ggtggattcc actgataagg
     1261 ctgatctgag actgatctac ctggctctgg ctcacatgat caagttcaga ggacacttcc
     1321 tgatcgaggg agatctgaac ccagataact ccgatgtgga taagctgttc atccaactgg
     1381 tgcaaactta caaccaactg ttcgaggaga acccaatcaa cgcttccgga gtggatgcta
     1441 aggctatcct gtccgctaga ctgtccaagt ccagaagact ggagaacctg atcgctcaac
     1501 tgccaggaga gaagaagaac ggactgttcg gaaacctgat cgctctgtcc ctgggactga
     1561 ctccaaactt caagtccaac ttcgatctgg ctgaggatgc taagctgcaa ctgtccaagg
     1621 atacttacga tgatgatctg gataacctgc tggctcaaat cggagatcaa tacgctgatc
     1681 tgttcctggc tgctaagaac ctgtccgatg ctatcctgct gtccgatatc ctgagagtga
     1741 acactgagat cactaaggct ccactgtccg cttccatgat caagagatac gatgagcacc
     1801 accaagatct gactctgctg aaggctctgg tgagacaaca actgccagag aagtacaagg
     1861 agatcttctt cgatcaatcc aagaacggat acgctggata catcgatgga ggagcttccc
     1921 aagaggagtt ctacaagttc atcaagccaa tcctggagaa gatggatgga actgaggagc
     1981 tgctggtgaa gctgaacaga gaggatctgc tgagaaagca aagaactttc gataacggat
     2041 ccatcccaca ccaaatccac ctgggagagc tgcacgctat cctgagaaga caagaggatt
     2101 tctacccatt cctgaaggat aacagagaga agatcgagaa gatcctgact ttcagaatcc
     2161 catactacgt gggaccactg gctagaggaa actccagatt cgcttggatg actagaaagt
     2221 ccgaggagac tatcactcca tggaacttcg aggaggtggt ggataaggga gcttccgctc
     2281 aatccttcat cgagagaatg actaacttcg ataagaacct gccaaacgag aaggtgctgc
     2341 caaagcactc cctgctgtac gagtacttca ctgtgtacaa cgagctgact aaggtgaagt
     2401 acgtgactga gggaatgaga aagccagctt tcctgtccgg agagcaaaag aaggctatcg
     2461 tggatctgct gttcaagact aacagaaagg tgactgtgaa gcaactgaag gaggattact
     2521 tcaagaagat cgagtgcttc gattccgtgg agatctccgg agtggaggat agattcaacg
     2581 cttccctggg aacttaccac gatctgctga agatcatcaa ggataaggat ttcctggata
     2641 acgaggagaa cgaggatatc ctggaggata tcgtgctgac tctgactctg ttcgaggata
     2701 gagagatgat cgaggagaga ctgaagactt acgctcacct gttcgatgat aaggtgatga
     2761 agcaactgaa gagaagaaga tacactggat ggggaagact gtccagaaag ctgatcaacg
     2821 gaatcagaga taagcaatcc ggaaagacta tcctggattt cctgaagtcc gatggattcg
     2881 ctaacagaaa cttcatgcaa ctgatccacg atgattccct gactttcaag gaggatatcc
     2941 aaaaggctca agtgtccgga caaggagatt ccctgcacga gcacatcgct aacctggctg
     3001 gatccccagc tatcaagaag ggaatcctgc aaactgtgaa ggtggtggat gagctggtga
     3061 aggtgatggg aagacacaag ccagagaaca tcgtgatcga gatggctaga gagaaccaaa
     3121 ctactcaaaa gggacaaaag aactccagag agagaatgaa gagaatcgag gagggaatca
     3181 aggagctggg atcccaaatc ctgaaggagc acccagtgga gaacactcaa ctgcaaaacg
     3241 agaagctgta cctgtactac ctgcaaaacg gaagagatat gtacgtggat caagagctgg
     3301 atatcaacag actgtccgat tacgatgtgg atcacatcgt gccacaatcc ttcctgaagg
     3361 atgattccat cgataacaag gtgctgacta gatccgataa gaacagagga aagtccgata
     3421 acgtgccatc cgaggaggtg gtgaagaaga tgaagaacta ctggagacaa ctgctgaacg
     3481 ctaagctgat cactcaaaga aagttcgata acctgactaa ggctgagaga ggaggactgt
     3541 ccgagctgga taaggctgga ttcatcaaga gacaactggt ggagactaga caaatcacta
     3601 agcacgtggc tcaaatcctg gattccagaa tgaacactaa gtacgatgag aacgataagc
     3661 tgatcagaga ggtgaaggtg atcactctga agtccaagct ggtgtccgat ttcagaaagg
     3721 atttccaatt ctacaaggtg agagagatca acaactacca ccacgctcac gatgcttacc
     3781 tgaacgctgt ggtgggaact gctctgatca agaagtaccc aaagctggag tccgagttcg
     3841 tgtacggaga ttacaaggtg tacgatgtga gaaagatgat cgctaagtcc gagcaagaga
     3901 tcggaaaggc tactgctaag tacttcttct actccaacat catgaacttc ttcaagactg
     3961 agatcactct ggctaacgga gagatcagaa agagaccact gatcgagact aacggagaga
     4021 ctggagagat cgtgtgggat aagggaagag atttcgctac tgtgagaaag gtgctgtcca
     4081 tgccacaagt gaacatcgtg aagaagactg aggtgcaaac tggaggattc tccaaggagt
     4141 ccatcctgcc aaagagaaac tccgataagc tgatcgctag aaagaaggat tgggatccaa
     4201 agaagtacgg aggattcgat tccccaactg tggcttactc cgtgctggtg gtggctaagg
     4261 tggagaaggg aaagtccaag aagctgaagt ccgtgaagga gctgctggga atcactatca
     4321 tggagagatc ctccttcgag aagaacccaa tcgatttcct ggaggctaag ggatacaagg
     4381 aggtgaagaa ggatctgatc atcaagctgc caaagtactc cctgttcgag ctggagaacg
     4441 gaagaaagag aatgctggct tccgctggag agctgcaaaa gggaaacgag ctggctctgc
     4501 catccaagta cgtgaacttc ctgtacctgg cttcccacta cgagaagctg aagggatccc
     4561 cagaggataa cgagcaaaag caactgttcg tggagcaaca caagcactac ctggatgaga
     4621 tcatcgagca aatctccgag ttctccaaga gagtgatcct ggctgatgct aacctggata
     4681 aggtgctgtc cgcttacaac aagcacagag ataagccaat cagagagcaa gctgagaaca
     4741 tcatccacct gttcactctg actaacctgg gagctccagc tgctttcaag tacttcgata
     4801 ctactatcga tagaaagaga tacacttcca ctaaggaggt gctggatgct actctgatcc
     4861 accaatccat cactggactg tacgagacta gaatcgatct gtcccaactg ggaggagatc
     4921 tgcaaccaaa gaagaagaga aaggtgggag gatgatctag accagattaa ttaactagtt
     4981 ctagagcggc cgccaccgcg gtggagctcg aatttccccg atcgttcaaa catttggcaa
     5041 taaagtttct taagattgaa tcctgttgcc ggtcttgcga tgattatcat ataatttctg
     5101 ttgaattacg ttaagcatgt aataattaac atgtaatgca tgacgttatt tatgagatgg
     5161 gtttttatga ttagagtccc gcaattatac atttaatacg cgatagaaaa caaaatatag
     5221 cgcgcaaact aggataaatt atcgcgcgcg gtgtcatcta tgttactcgt aatcatgtca
     5281 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     5341 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     5401 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     5461 caacgcgcgg ggagaggcgg tttgcgtatt ggctagagca gcttgccaac atggtggagc
     5521 acgacactct cgtctactcc aagaatatca aagatacagt ctcagaagac caaagggcta
     5581 ttgagacttt tcaacaaagg gtaatatcgg gaaacctcct cggattccat tgcccagcta
     5641 tctgtcactt catcaaaagg acagtagaaa aggaaggtgg cacctacaaa tgccatcatt
     5701 gcgataaagg aaaggctatc gttcaagatg cctctgccga cagtggtccc aaagatggac
     5761 ccccacccac gaggagcatc gtggaaaaag aagacgttcc aaccacgtct tcaaagcaag
     5821 tggattgatg tgaacatggt ggagcacgac actctcgtct actccaagaa tatcaaagat
     5881 acagtctcag aagaccaaag ggctattgag acttttcaac aaagggtaat atcgggaaac
     5941 ctcctcggat tccattgccc agctatctgt cacttcatca aaaggacagt agaaaaggaa
     6001 ggtggcacct acaaatgcca tcattgcgat aaaggaaagg ctatcgttca agatgcctct
     6061 gccgacagtg gtcccaaaga tggaccccca cccacgagga gcatcgtgga aaaagaagac
     6121 gttccaacca cgtcttcaaa gcaagtggat tgatgtgata tctccactga cgtaagggat
     6181 gacgcacaat cccactatcc ttcgcaagac ccttcctcta tataaggaag ttcatttcat
     6241 ttggagagga cacgctgaaa tcaccagtct ctctctacaa atctatctct ctcgagcttt
     6301 cgcagatccg gggggcaatg agatatgaaa aagcctgaac tcaccgcgac gtctgtcgag
     6361 aagtttctga tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc ggagggcgaa
     6421 gaatctcgtg ctttcagctt cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc
     6481 tgcgccgatg gtttctacaa agatcgttat gtttatcggc actttgcatc ggccgcgctc
     6541 ccgattccgg aagtgcttga cattggggag tttagcgaga gcctgaccta ttgcatctcc
     6601 cgccgttcac agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc cgctgttcta
     6661 caaccggtcg cggaggctat ggatgcgatc gctgcggccg atcttagcca gacgagcggg
     6721 ttcggcccat tcggaccgca aggaatcggt caatacacta catggcgtga tttcatatgc
     6781 gcgattgctg atccccatgt gtatcactgg caaactgtga tggacgacac cgtcagtgcg
     6841 tccgtcgcgc aggctctcga tgagctgatg ctttgggccg aggactgccc cgaagtccgg
     6901 cacctcgtgc acgcggattt cggctccaac aatgtcctga cggacaatgg ccgcataaca
     6961 gcggtcattg actggagcga ggcgatgttc ggggattccc aatacgaggt cgccaacatc
     7021 ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt cgagcggagg
     7081 catccggagc ttgcaggatc gccacgactc cgggcgtata tgctccgcat tggtcttgac
     7141 caactctatc agagcttggt tgacggcaat ttcgatgatg cagcttgggc gcagggtcga
     7201 tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc gtacacaaat cgcccgcaga
     7261 agcgcggccg tctggaccga tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc
     7321 cccagcactc gtccgagggc aaagaaatag agtagatgcc gaccgggatc tgtcgatcga
     7381 caagctcgag tttctccata ataatgtgtg agtagttccc agataaggga attagggttc
     7441 ctatagggtt tcgctcatgt gttgagcata taagaaaccc ttagtatgta tttgtatttg
     7501 taaaatactt ctatcaataa aatttctaat tcctaaaacc aaaatccagt actaaaatcc
     7561 agatcccccg aattaattcg gcgttaattc agtacattaa aaacgtccgc aatgtgttat
     7621 taagttgtct aagcgtcaat ttgtttacac cacaatatat cctgccacca gccagccaac
     7681 agctccccga ccggcagctc ggcacaaaat caccactcga tacaggcagc ccatcagtcc
     7741 gggacggcgt cagcgggaga gccgttgtaa ggcggcagac tttgctcatg ttaccgatgc
     7801 tattcggaag aacggcaact aagctgccgg gtttgaaaca cggatgatct cgcggagggt
     7861 agcatgttga ttgtaacgat gacagagcgt tgctgcctgt gatcaccgcg gtttcaaaat
     7921 cggctccgtc gatactatgt tatacgccaa ctttgaaaac aactttgaaa aagctgtttt
     7981 ctggtattta aggttttaga atgcaaggaa cagtgaattg gagttcgtct tgttataatt
     8041 agggaattgg ggtatcttta aatactgtag aaaagaggaa ggaaataata aatggctaaa
     8101 atgagaatat caccggaatt gaaaaaactg atcgaaaaat accgctgcgt aaaagatacg
     8161 gaaggaatgt ctcctgctaa ggtatataag ctggtgggag aaaatgaaaa cctatattta
     8221 aaaatgacgg acagccggta taaagggacc acctatgatg tggaacggga aaaggacatg
     8281 atgctatggc tggaaggaaa gctgcctgtt ccaaaggtcc tgcactttga acggcatgat
     8341 ggctggagca atctgctcat gagtgaggcc gatggcgtcc tttgctcgga agagtatgaa
     8401 gatgaacaaa gccctgaaaa gattatcgag ctgtatgcgg agtgcatcag gctctttcac
     8461 tccatcgaca tatcggattg tccctatacg aatagcttag acagccgctt agccgaattg
     8521 gattacttac tgaataacga tctggccgat gtggattgcg aaaactggga agaagacact
     8581 ccatttaaag atccgcgcga gctgtatgat tttttaaaga cggaaaagcc cgaagaggaa
     8641 cttgtctttt cccacggcga cctgggagac agcaacatct ttgtgaaaga tggcaaagta
     8701 agtggcttta ttgatcttgg gagaagcggc agggcggaca agtggtatga cattgccttc
     8761 tgcgtccggt cgatcaggga ggatatcggg gaagaacagt atgtcgagct attttttgac
     8821 ttactgggga tcaagcctga ttgggagaaa ataaaatatt atattttact ggatgaattg
     8881 ttttagtacc tagaatgcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     8941 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     9001 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     9061 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     9121 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     9181 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     9241 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     9301 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     9361 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     9421 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     9481 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     9541 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     9601 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     9661 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     9721 tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc
     9781 ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     9841 agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc
     9901 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     9961 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
    10021 cgaggcaggg tgccttgatg tgggcgccgg cggtcgagtg gcgacggcgc ggcttgtccg
    10081 cgccctggta gattgcctgg ccgtaggcca gccatttttg agcggccagc ggccgcgata
    10141 ggccgacgcg aagcggcggg gcgtagggag cgcagcgacc gaagggtagg cgctttttgc
    10201 agctcttcgg ctgtgcgctg gccagacagt tatgcacagg ccaggcgggt tttaagagtt
    10261 ttaataagtt ttaaagagtt ttaggcggaa aaatcgcctt ttttctcttt tatatcagtc
    10321 acttacatgt gtgaccggtt cccaatgtac ggctttgggt tcccaatgta cgggttccgg
    10381 ttcccaatgt acggctttgg gttcccaatg tacgtgctat ccacaggaaa gagacctttt
    10441 cgaccttttt cccctgctag ggcaatttgc cctagcatct gctccgtaca ttaggaaccg
    10501 gcggatgctt cgccctcgat caggttgcgg tagcgcatga ctaggatcgg gccagcctgc
    10561 cccgcctcct ccttcaaatc gtactccggc aggtcatttg acccgatcag cttgcgcacg
    10621 gtgaaacaga acttcttgaa ctctccggcg ctgccactgc gttcgtagat cgtcttgaac
    10681 aaccatctgg cttctgcctt gcctgcggcg cggcgtgcca ggcggtagag aaaacggccg
    10741 atgccgggat cgatcaaaaa gtaatcgggg tgaaccgtca gcacgtccgg gttcttgcct
    10801 tctgtgatct cgcggtacat ccaatcagct agctcgatct cgatgtactc cggccgcccg
    10861 gtttcgctct ttacgatctt gtagcggcta atcaaggctt caccctcgga taccgtcacc
    10921 aggcggccgt tcttggcctt cttcgtacgc tgcatggcaa cgtgcgtggt gtttaaccga
    10981 atgcaggttt ctaccaggtc gtctttctgc tttccgccat cggctcgccg gcagaacttg
    11041 agtacgtccg caacgtgtgg acggaacacg cggccgggct tgtctccctt cccttcccgg
    11101 tatcggttca tggattcggt tagatgggaa accgccatca gtaccaggtc gtaatcccac
    11161 acactggcca tgccggccgg ccctgcggaa acctctacgt gcccgtctgg aagctcgtag
    11221 cggatcacct cgccagctcg tcggtcacgc ttcgacagac ggaaaacggc cacgtccatg
    11281 atgctgcgac tatcgcgggt gcccacgtca tagagcatcg gaacgaaaaa atctggttgc
    11341 tcgtcgccct tgggcggctt cctaatcgac ggcgcaccgg ctgccggcgg ttgccgggat
    11401 tctttgcgga ttcgatcagc ggccgcttgc cacgattcac cggggcgtgc ttctgcctcg
    11461 atgcgttgcc gctgggcggc ctgcgcggcc ttcaacttct ccaccaggtc atcacccagc
    11521 gccgcgccga tttgtaccgg gccggatggt ttgcgaccgc tcacgccgat tcctcgggct
    11581 tgggggttcc agtgccattg cagggccggc aggcaaccca gccgcttacg cctggccaac
    11641 cgcccgttcc tccacacatg gggcattcca cggcgtcggt gcctggttgt tcttgatttt
    11701 ccatgccgcc tcctttagcc gctaaaattc atctactcat ttattcattt gctcatttac
    11761 tctggtagct gcgcgatgta ttcagatagc agctcggtaa tggtcttgcc ttggcgtacc
    11821 gcgtacatct tcagcttggt gtgatcctcc gccggcaact gaaagttgac ccgcttcatg
    11881 gctggcgtgt ctgccaggct ggccaacgtt gcagccttgc tgctgcgtgc gctcggacgg
    11941 ccggcactta gcgtgtttgt gcttttgctc attttctctt tacctcatta actcaaatga
    12001 gttttgattt aatttcagcg gccagcgcct ggacctcgcg ggcagcgtcg ccctcgggtt
    12061 ctgattcaag aacggttgtg ccggcggcgg cagtgcctgg gtagctcacg cgctgcgtga
    12121 tacgggactc aagaatgggc agctcgtacc cggccagcgc ctcggcaacc tcaccgccga
    12181 tgcgcgtgcc tttgatcgcc cgcgacacga caaaggccgc ttgtagcctt ccatccgtga
    12241 cctcaatgcg ctgcttaacc agctccacca ggtcggcggt ggcccatatg tcgtaagggc
    12301 ttggctgcac cggaatcagc acgaagtcgg ctgccttgat cgcggacaca gccaagtccg
    12361 ccgcctgggg cgctccgtcg atcactacga agtcgcgccg gccgatggcc ttcacgtcgc
    12421 ggtcaatcgt cgggcggtcg atgccgacaa cggttagcgg ttgatcttcc cgcacggccg
    12481 cccaatcgcg ggcactgccc tggggatcgg aatcgactaa cagaacatcg gccccggcga
    12541 gttgcagggc gcgggctaga tgggttgcga tggtcgtctt gcctgacccg cctttctggt
    12601 taagtacagc gataaccttc atgcgttccc cttgcgtatt tgtttattta ctcatcgcat
    12661 catatacgca gcgaccgcat gacgcaagct gttttactca aatacacatc acctttttag
    12721 acggcggcgc tcggtttctt cagcggccaa gctggccggc caggccgcca gcttggcatc
    12781 agacaaaccg gccaggattt catgcagccg cacggttgag acgtgcgcgg gcggctcgaa
    12841 cacgtacccg gccgcgatca tctccgcctc gatctcttcg gtaatgaaaa acggttcgtc
    12901 ctggccgtcc tggtgcggtt tcatgcttgt tcctcttggc gttcattctc ggcggccgcc
    12961 agggcgtcgg cctcggtcaa tgcgtcctca cggaaggcac cgcgccgcct ggcctcggtg
    13021 ggcgtcactt cctcgctgcg ctcaagtgcg cggtacaggg tcgagcgatg cacgccaagc
    13081 agtgcagccg cctctttcac ggtgcggcct tcctggtcga tcagctcgcg ggcgtgcgcg
    13141 atctgtgccg gggtgagggt agggcggggg ccaaacttca cgcctcgggc cttggcggcc
    13201 tcgcgcccgc tccgggtgcg gtcgatgatt agggaacgct cgaactcggc aatgccggcg
    13261 aacacggtca acaccatgcg gccggccggc gtggtggtgt cggcccacgg ctctgccagg
    13321 ctacgcaggc ccgcgccggc ctcctggatg cgctcggcaa tgtccagtag gtcgcgggtg
    13381 ctgcgggcca ggcggtctag cctggtcact gtcacaacgt cgccagggcg taggtggtca
    13441 agcatcctgg ccagctccgg gcggtcgcgc ctggtgccgg tgatcttctc ggaaaacagc
    13501 ttggtgcagc cggccgcgtg cagttcggcc cgttggttgg tcaagtcctg gtcgtcggtg
    13561 ctgacgcggg catagcccag caggccagcg gcggcgctct tgttcatggc gtaatgtctc
    13621 cggttctagt cgcaagtatt ctactttatg cgactaaaac acgcgacaag aaaacgccag
    13681 gaaaagggca gggcggcagc ctgtcgcgta acttaggact tgtgcgacat gtcgttttca
    13741 gaagacggct gcactgaacg tcagaagccg actgca
//
