LOCUS       sequence-169048-       13204 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  G10 promoting Glycing Max codon optimized Cas9 (N and C terminus 
            NLS) with bar plant selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13204)
  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 13204)
  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: G10 Cas9 MDC123
FEATURES             Location/Qualifiers
     source          1..13204
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    5..145
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(71..90)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     231..253
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      489..683
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(749..1822)
                     /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(2251..2880)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     misc_feature    4181..4205
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     4393..4415
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4407..4424
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4408..4424
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4689..4735
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             4761..4781
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4788..8891
                     /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             4788..8891
                     /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             8804..8812
                     /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             8898..8918
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      8957..9209
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(9269..9285)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9269..9285)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9282..9304)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9293..9309
                     /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(9317..9323,9324..9341,9342..9347))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9362..9383
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        9574..10250
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     CDS             10295..10846
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     polyA_signal    10853..11027
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    11105..11129
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             11554..12348
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      12435..13023
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12924..12943
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     13177..13194
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt
       61 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc
      121 gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg
      181 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
      241 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagggtg ccttgatgtg
      301 ggcgccggcg gtcgagtggc gacggcgcgg cttgtccgcg ccctggtaga ttgcctggcc
      361 gtaggccagc catttttgag cggccagcgg ccgcgatagg ccgacgcgaa gcggcggggc
      421 gtagggagcg cagcgaccga agggtaggcg ctttttgcag ctcttcggct gtgcgctggc
      481 cagacagtta tgcacaggcc aggcgggttt taagagtttt aataagtttt aaagagtttt
      541 aggcggaaaa atcgcctttt ttctctttta tatcagtcac ttacatgtgt gaccggttcc
      601 caatgtacgg ctttgggttc ccaatgtacg ggttccggtt cccaatgtac ggctttgggt
      661 tcccaatgta cgtgctatcc acaggaaaga gaccttttcg acctttttcc cctgctaggg
      721 caatttgccc tagcatctgc tccgtacatt aggaaccggc ggatgcttcg ccctcgatca
      781 ggttgcggta gcgcatgact aggatcgggc cagcctgccc cgcctcctcc ttcaaatcgt
      841 actccggcag gtcatttgac ccgatcagct tgcgcacggt gaaacagaac ttcttgaact
      901 ctccggcgct gccactgcgt tcgtagatcg tcttgaacaa ccatctggct tctgccttgc
      961 ctgcggcgcg gcgtgccagg cggtagagaa aacggccgat gccgggatcg atcaaaaagt
     1021 aatcggggtg aaccgtcagc acgtccgggt tcttgccttc tgtgatctcg cggtacatcc
     1081 aatcagctag ctcgatctcg atgtactccg gccgcccggt ttcgctcttt acgatcttgt
     1141 agcggctaat caaggcttca ccctcggata ccgtcaccag gcggccgttc ttggccttct
     1201 tcgtacgctg catggcaacg tgcgtggtgt ttaaccgaat gcaggtttct accaggtcgt
     1261 ctttctgctt tccgccatcg gctcgccggc agaacttgag tacgtccgca acgtgtggac
     1321 ggaacacgcg gccgggcttg tctcccttcc cttcccggta tcggttcatg gattcggtta
     1381 gatgggaaac cgccatcagt accaggtcgt aatcccacac actggccatg ccggccggcc
     1441 ctgcggaaac ctctacgtgc ccgtctggaa gctcgtagcg gatcacctcg ccagctcgtc
     1501 ggtcacgctt cgacagacgg aaaacggcca cgtccatgat gctgcgacta tcgcgggtgc
     1561 ccacgtcata gagcatcgga acgaaaaaat ctggttgctc gtcgcccttg ggcggcttcc
     1621 taatcgacgg cgcaccggct gccggcggtt gccgggattc tttgcggatt cgatcagcgg
     1681 ccgcttgcca cgattcaccg gggcgtgctt ctgcctcgat gcgttgccgc tgggcggcct
     1741 gcgcggcctt caacttctcc accaggtcat cacccagcgc cgcgccgatt tgtaccgggc
     1801 cggatggttt gcgaccgctc acgccgattc ctcgggcttg ggggttccag tgccattgca
     1861 gggccggcag gcaacccagc cgcttacgcc tggccaaccg cccgttcctc cacacatggg
     1921 gcattccacg gcgtcggtgc ctggttgttc ttgattttcc atgccgcctc ctttagccgc
     1981 taaaattcat ctactcattt attcatttgc tcatttactc tggtagctgc gcgatgtatt
     2041 cagatagcag ctcggtaatg gtcttgcctt ggcgtaccgc gtacatcttc agcttggtgt
     2101 gatcctccgc cggcaactga aagttgaccc gcttcatggc tggcgtgtct gccaggctgg
     2161 ccaacgttgc agccttgctg ctgcgtgcgc tcggacggcc ggcacttagc gtgtttgtgc
     2221 ttttgctcat tttctcttta cctcattaac tcaaatgagt tttgatttaa tttcagcggc
     2281 cagcgcctgg acctcgcggg cagcgtcgcc ctcgggttct gattcaagaa cggttgtgcc
     2341 ggcggcggca gtgcctgggt agctcacgcg ctgcgtgata cgggactcaa gaatgggcag
     2401 ctcgtacccg gccagcgcct cggcaacctc accgccgatg cgcgtgcctt tgatcgcccg
     2461 cgacacgaca aaggccgctt gtagccttcc atccgtgacc tcaatgcgct gcttaaccag
     2521 ctccaccagg tcggcggtgg cccatatgtc gtaagggctt ggctgcaccg gaatcagcac
     2581 gaagtcggct gccttgatcg cggacacagc caagtccgcc gcctggggcg ctccgtcgat
     2641 cactacgaag tcgcgccggc cgatggcctt cacgtcgcgg tcaatcgtcg ggcggtcgat
     2701 gccgacaacg gttagcggtt gatcttcccg cacggccgcc caatcgcggg cactgccctg
     2761 gggatcggaa tcgactaaca gaacatcggc cccggcgagt tgcagggcgc gggctagatg
     2821 ggttgcgatg gtcgtcttgc ctgacccgcc tttctggtta agtacagcga taaccttcat
     2881 gcgttcccct tgcgtatttg tttatttact catcgcatca tatacgcagc gaccgcatga
     2941 cgcaagctgt tttactcaaa tacacatcac ctttttagac ggcggcgctc ggtttcttca
     3001 gcggccaagc tggccggcca ggccgccagc ttggcatcag acaaaccggc caggatttca
     3061 tgcagccgca cggttgagac gtgcgcgggc ggctcgaaca cgtacccggc cgcgatcatc
     3121 tccgcctcga tctcttcggt aatgaaaaac ggttcgtcct ggccgtcctg gtgcggtttc
     3181 atgcttgttc ctcttggcgt tcattctcgg cggccgccag ggcgtcggcc tcggtcaatg
     3241 cgtcctcacg gaaggcaccg cgccgcctgg cctcggtggg cgtcacttcc tcgctgcgct
     3301 caagtgcgcg gtacagggtc gagcgatgca cgccaagcag tgcagccgcc tctttcacgg
     3361 tgcggccttc ctggtcgatc agctcgcggg cgtgcgcgat ctgtgccggg gtgagggtag
     3421 ggcgggggcc aaacttcacg cctcgggcct tggcggcctc gcgcccgctc cgggtgcggt
     3481 cgatgattag ggaacgctcg aactcggcaa tgccggcgaa cacggtcaac accatgcggc
     3541 cggccggcgt ggtggtgtcg gcccacggct ctgccaggct acgcaggccc gcgccggcct
     3601 cctggatgcg ctcggcaatg tccagtaggt cgcgggtgct gcgggccagg cggtctagcc
     3661 tggtcactgt cacaacgtcg ccagggcgta ggtggtcaag catcctggcc agctccgggc
     3721 ggtcgcgcct ggtgccggtg atcttctcgg aaaacagctt ggtgcagccg gccgcgtgca
     3781 gttcggcccg ttggttggtc aagtcctggt cgtcggtgct gacgcgggca tagcccagca
     3841 ggccagcggc ggcgctcttg ttcatggcgt aatgtctccg gttctagtcg caagtattct
     3901 actttatgcg actaaaacac gcgacaagaa aacgccagga aaagggcagg gcggcagcct
     3961 gtcgcgtaac ttaggacttg tgcgacatgt cgttttcaga agacggctgc actgaacgtc
     4021 agaagccgac tgcactatag cagcggaggg gttggatcaa agtactttga tcccgagggg
     4081 aaccctgtgg ttggcatgca catacaaatg gacgaacgga taaacctttt cacgcccttt
     4141 taaatatccg attattctaa taaacgctct tttctcttag gtttacccgc caatatatcc
     4201 tgtcaaacac tgatagttta aactgaaggc gggaaacgac aatctgatcc aagctcaagc
     4261 tgctctagca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     4321 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     4381 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgccaag cttgcatgcc
     4441 tgcaggtcga ctctagagga tccccgggta ccgggccccc cctcgaggcg cgccaccagg
     4501 tcgacatagt ttaaactgaa ggcgggaaac gacaatctga tccaagctca agctaagctt
     4561 gcatgcctgc aggatatcgt ggatccaagc ttgccacgtg ccgccacgtg ccgccacgtg
     4621 ccgccacgtg cctctagagg atccatctcc actgacgtaa gggatgacgc acaatcccac
     4681 tatccttcgc aagacccttc ctctatataa ggaagttcat ttcatttgga gaggacacgc
     4741 tgcttatgtc cagagctgat ccaaagaaga agagaaaggt ggtcgacatg gataagaagt
     4801 actccatcgg actggatatc ggaactaact ccgtgggatg ggctgtgatc actgatgagt
     4861 acaaggtgcc atccaagaag ttcaaggtgc tgggaaacac tgatagacac tccatcaaga
     4921 agaacctgat cggagctctg ctgttcgatt ccggagagac tgctgaggct actagactga
     4981 agagaactgc tagaagaaga tacactagaa gaaagaacag aatctgctac ctgcaagaga
     5041 tcttctccaa cgagatggct aaggtggatg attccttctt ccacagactg gaggagtcct
     5101 tcctggtgga ggaggataag aagcacgaga gacacccaat cttcggaaac atcgtggatg
     5161 aggtggctta ccacgagaag tacccaacta tctaccacct gagaaagaag ctggtggatt
     5221 ccactgataa ggctgatctg agactgatct acctggctct ggctcacatg atcaagttca
     5281 gaggacactt cctgatcgag ggagatctga acccagataa ctccgatgtg gataagctgt
     5341 tcatccaact ggtgcaaact tacaaccaac tgttcgagga gaacccaatc aacgcttccg
     5401 gagtggatgc taaggctatc ctgtccgcta gactgtccaa gtccagaaga ctggagaacc
     5461 tgatcgctca actgccagga gagaagaaga acggactgtt cggaaacctg atcgctctgt
     5521 ccctgggact gactccaaac ttcaagtcca acttcgatct ggctgaggat gctaagctgc
     5581 aactgtccaa ggatacttac gatgatgatc tggataacct gctggctcaa atcggagatc
     5641 aatacgctga tctgttcctg gctgctaaga acctgtccga tgctatcctg ctgtccgata
     5701 tcctgagagt gaacactgag atcactaagg ctccactgtc cgcttccatg atcaagagat
     5761 acgatgagca ccaccaagat ctgactctgc tgaaggctct ggtgagacaa caactgccag
     5821 agaagtacaa ggagatcttc ttcgatcaat ccaagaacgg atacgctgga tacatcgatg
     5881 gaggagcttc ccaagaggag ttctacaagt tcatcaagcc aatcctggag aagatggatg
     5941 gaactgagga gctgctggtg aagctgaaca gagaggatct gctgagaaag caaagaactt
     6001 tcgataacgg atccatccca caccaaatcc acctgggaga gctgcacgct atcctgagaa
     6061 gacaagagga tttctaccca ttcctgaagg ataacagaga gaagatcgag aagatcctga
     6121 ctttcagaat cccatactac gtgggaccac tggctagagg aaactccaga ttcgcttgga
     6181 tgactagaaa gtccgaggag actatcactc catggaactt cgaggaggtg gtggataagg
     6241 gagcttccgc tcaatccttc atcgagagaa tgactaactt cgataagaac ctgccaaacg
     6301 agaaggtgct gccaaagcac tccctgctgt acgagtactt cactgtgtac aacgagctga
     6361 ctaaggtgaa gtacgtgact gagggaatga gaaagccagc tttcctgtcc ggagagcaaa
     6421 agaaggctat cgtggatctg ctgttcaaga ctaacagaaa ggtgactgtg aagcaactga
     6481 aggaggatta cttcaagaag atcgagtgct tcgattccgt ggagatctcc ggagtggagg
     6541 atagattcaa cgcttccctg ggaacttacc acgatctgct gaagatcatc aaggataagg
     6601 atttcctgga taacgaggag aacgaggata tcctggagga tatcgtgctg actctgactc
     6661 tgttcgagga tagagagatg atcgaggaga gactgaagac ttacgctcac ctgttcgatg
     6721 ataaggtgat gaagcaactg aagagaagaa gatacactgg atggggaaga ctgtccagaa
     6781 agctgatcaa cggaatcaga gataagcaat ccggaaagac tatcctggat ttcctgaagt
     6841 ccgatggatt cgctaacaga aacttcatgc aactgatcca cgatgattcc ctgactttca
     6901 aggaggatat ccaaaaggct caagtgtccg gacaaggaga ttccctgcac gagcacatcg
     6961 ctaacctggc tggatcccca gctatcaaga agggaatcct gcaaactgtg aaggtggtgg
     7021 atgagctggt gaaggtgatg ggaagacaca agccagagaa catcgtgatc gagatggcta
     7081 gagagaacca aactactcaa aagggacaaa agaactccag agagagaatg aagagaatcg
     7141 aggagggaat caaggagctg ggatcccaaa tcctgaagga gcacccagtg gagaacactc
     7201 aactgcaaaa cgagaagctg tacctgtact acctgcaaaa cggaagagat atgtacgtgg
     7261 atcaagagct ggatatcaac agactgtccg attacgatgt ggatcacatc gtgccacaat
     7321 ccttcctgaa ggatgattcc atcgataaca aggtgctgac tagatccgat aagaacagag
     7381 gaaagtccga taacgtgcca tccgaggagg tggtgaagaa gatgaagaac tactggagac
     7441 aactgctgaa cgctaagctg atcactcaaa gaaagttcga taacctgact aaggctgaga
     7501 gaggaggact gtccgagctg gataaggctg gattcatcaa gagacaactg gtggagacta
     7561 gacaaatcac taagcacgtg gctcaaatcc tggattccag aatgaacact aagtacgatg
     7621 agaacgataa gctgatcaga gaggtgaagg tgatcactct gaagtccaag ctggtgtccg
     7681 atttcagaaa ggatttccaa ttctacaagg tgagagagat caacaactac caccacgctc
     7741 acgatgctta cctgaacgct gtggtgggaa ctgctctgat caagaagtac ccaaagctgg
     7801 agtccgagtt cgtgtacgga gattacaagg tgtacgatgt gagaaagatg atcgctaagt
     7861 ccgagcaaga gatcggaaag gctactgcta agtacttctt ctactccaac atcatgaact
     7921 tcttcaagac tgagatcact ctggctaacg gagagatcag aaagagacca ctgatcgaga
     7981 ctaacggaga gactggagag atcgtgtggg ataagggaag agatttcgct actgtgagaa
     8041 aggtgctgtc catgccacaa gtgaacatcg tgaagaagac tgaggtgcaa actggaggat
     8101 tctccaagga gtccatcctg ccaaagagaa actccgataa gctgatcgct agaaagaagg
     8161 attgggatcc aaagaagtac ggaggattcg attccccaac tgtggcttac tccgtgctgg
     8221 tggtggctaa ggtggagaag ggaaagtcca agaagctgaa gtccgtgaag gagctgctgg
     8281 gaatcactat catggagaga tcctccttcg agaagaaccc aatcgatttc ctggaggcta
     8341 agggatacaa ggaggtgaag aaggatctga tcatcaagct gccaaagtac tccctgttcg
     8401 agctggagaa cggaagaaag agaatgctgg cttccgctgg agagctgcaa aagggaaacg
     8461 agctggctct gccatccaag tacgtgaact tcctgtacct ggcttcccac tacgagaagc
     8521 tgaagggatc cccagaggat aacgagcaaa agcaactgtt cgtggagcaa cacaagcact
     8581 acctggatga gatcatcgag caaatctccg agttctccaa gagagtgatc ctggctgatg
     8641 ctaacctgga taaggtgctg tccgcttaca acaagcacag agataagcca atcagagagc
     8701 aagctgagaa catcatccac ctgttcactc tgactaacct gggagctcca gctgctttca
     8761 agtacttcga tactactatc gatagaaaga gatacacttc cactaaggag gtgctggatg
     8821 ctactctgat ccaccaatcc atcactggac tgtacgagac tagaatcgat ctgtcccaac
     8881 tgggaggaga tctgcaacca aagaagaaga gaaaggtggg aggatgatct agaccagatt
     8941 aattaagaat ttccccgatc gttcaaacat ttggcaataa agtttcttaa gattgaatcc
     9001 tgttgccggt cttgcgatga ttatcatata atttctgttg aattacgtta agcatgtaat
     9061 aattaacatg taatgcatga cgttatttat gagatgggtt tttatgatta gagtcccgca
     9121 attatacatt taatacgcga tagaaaacaa aatatagcgc gcaaactagg ataaattatc
     9181 gcgcgcggtg tcatctatgt tactagatcg gttaattaac tagttctaga gcggccgcca
     9241 ccgcggtgga gctcgaattc gtaatcatgt catagctgtt tcctgtgtga aattgttatc
     9301 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     9361 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     9421 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     9481 ttggctagag cagcttgcca acatggtgga gcacgacact ctcgtctact ccaagaatat
     9541 caaagataca gtctcagaag accaaagggc tattgagact tttcaacaaa gggtaatatc
     9601 gggaaacctc ctcggattcc attgcccagc tatctgtcac ttcatcaaaa ggacagtaga
     9661 aaaggaaggt ggcacctaca aatgccatca ttgcgataaa ggaaaggcta tcgttcaaga
     9721 tgcctctgcc gacagtggtc ccaaagatgg acccccaccc acgaggagca tcgtggaaaa
     9781 agaagacgtt ccaaccacgt cttcaaagca agtggattga tgtgaacatg gtggagcacg
     9841 acactctcgt ctactccaag aatatcaaag atacagtctc agaagaccaa agggctattg
     9901 agacttttca acaaagggta atatcgggaa acctcctcgg attccattgc ccagctatct
     9961 gtcacttcat caaaaggaca gtagaaaagg aaggtggcac ctacaaatgc catcattgcg
    10021 ataaaggaaa ggctatcgtt caagatgcct ctgccgacag tggtcccaaa gatggacccc
    10081 cacccacgag gagcatcgtg gaaaaagaag acgttccaac cacgtcttca aagcaagtgg
    10141 attgatgtga tatctccact gacgtaaggg atgacgcaca atcccactat ccttcgcaag
    10201 acccttcctc tatataagga agttcatttc atttggagag gacacgctga aatcaccagt
    10261 ctctctctac aaatctatct ctctcgagtc taccatgagc ccagaacgac gcccggccga
    10321 catccgccgt gccaccgagg cggacatgcc ggcggtctgc accatcgtca accactacat
    10381 cgagacaagc acggtcaact tccgtaccga gccgcaggaa ccgcaggagt ggacggacga
    10441 cctcgtccgt ctgcgggagc gctatccctg gctcgtcgcc gaggtggacg gcgaggtcgc
    10501 cggcatcgcc tacgcgggcc cctggaaggc acgcaacgcc tacgactgga cggccgagtc
    10561 gaccgtgtac gtctcccccc gccaccagcg gacgggactg ggctccacgc tctacaccca
    10621 cctgctgaag tccctggagg cacagggctt caagagcgtg gtcgctgtca tcgggctgcc
    10681 caacgacccg agcgtgcgca tgcacgaggc gctcggatat gccccccgcg gcatgctgcg
    10741 ggcggccggc ttcaagcacg ggaactggca tgacgtgggt ttctggcagc tggacttcag
    10801 cctgccggta ccgccccgtc cggtcctgcc cgtcaccgag atttgactcg agtttctcca
    10861 taataatgtg tgagtagttc ccagataagg gaattagggt tcctataggg tttcgctcat
    10921 gtgttgagca tataagaaac ccttagtatg tatttgtatt tgtaaaatac ttctatcaat
    10981 aaaatttcta attcctaaaa ccaaaatcca gtactaaaat ccagatcccc cgaattaatt
    11041 cggcgttaat tcagtacatt aaaaacgtcc gcaatgtgtt attaagttgt ctaagcgtca
    11101 atttgtttac accacaatat atcctgccac cagccagcca acagctcccc gaccggcagc
    11161 tcggcacaaa atcaccactc gatacaggca gcccatcagt ccgggacggc gtcagcggga
    11221 gagccgttgt aaggcggcag actttgctca tgttaccgat gctattcgga agaacggcaa
    11281 ctaagctgcc gggtttgaaa cacggatgat ctcgcggagg gtagcatgtt gattgtaacg
    11341 atgacagagc gttgctgcct gtgatcaccg cggtttcaaa atcggctccg tcgatactat
    11401 gttatacgcc aactttgaaa acaactttga aaaagctgtt ttctggtatt taaggtttta
    11461 gaatgcaagg aacagtgaat tggagttcgt cttgttataa ttagcttctt ggggtatctt
    11521 taaatactgt agaaaagagg aaggaaataa taaatggcta aaatgagaat atcaccggaa
    11581 ttgaaaaaac tgatcgaaaa ataccgctgc gtaaaagata cggaaggaat gtctcctgct
    11641 aaggtatata agctggtggg agaaaatgaa aacctatatt taaaaatgac ggacagccgg
    11701 tataaaggga ccacctatga tgtggaacgg gaaaaggaca tgatgctatg gctggaagga
    11761 aagctgcctg ttccaaaggt cctgcacttt gaacggcatg atggctggag caatctgctc
    11821 atgagtgagg ccgatggcgt cctttgctcg gaagagtatg aagatgaaca aagccctgaa
    11881 aagattatcg agctgtatgc ggagtgcatc aggctctttc actccatcga catatcggat
    11941 tgtccctata cgaatagctt agacagccgc ttagccgaat tggattactt actgaataac
    12001 gatctggccg atgtggattg cgaaaactgg gaagaagaca ctccatttaa agatccgcgc
    12061 gagctgtatg attttttaaa gacggaaaag cccgaagagg aacttgtctt ttcccacggc
    12121 gacctgggag acagcaacat ctttgtgaaa gatggcaaag taagtggctt tattgatctt
    12181 gggagaagcg gcagggcgga caagtggtat gacattgcct tctgcgtccg gtcgatcagg
    12241 gaggatatcg gggaagaaca gtatgtcgag ctattttttg acttactggg gatcaagcct
    12301 gattgggaga aaataaaata ttatatttta ctggatgaat tgttttagta cctagaatgc
    12361 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
    12421 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
    12481 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
    12541 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
    12601 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
    12661 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
    12721 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
    12781 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
    12841 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
    12901 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
    12961 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
    13021 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
    13081 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
    13141 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
    13201 gaag
//
