LOCUS       sequence-393227-       13663 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  2x35S promoting Glycing Max codon optimized Cas9 with bar plant 
            selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13663)
  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 13663)
  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 MDC123
FEATURES             Location/Qualifiers
     source          1..13663
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    83..107
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     295..317
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     309..326
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     310..326
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        752..1096
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             1157..5260
                     /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             1157..5260
                     /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             5173..5181
                     /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             5267..5287
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      5361..5613
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(5630..5646)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5630..5646)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5643..5665)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5654..5670
                     /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(5678..5684,5685..5702,5703..5708))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5723..5744
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5935..6611
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     CDS             6656..7207
                     /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    7214..7388
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    7466..7490
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             7915..8709
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      8796..9384
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9285..9304
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9538..9555
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    9570..9710
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(9636..9655)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9796..9818
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      10054..10248
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(10314..11387)
                     /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(11816..12445)
                     /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 cacatacaaa tggacgaacg gataaacctt ttcacgccct tttaaatatc cgattattct
       61 aataaacgct cttttctctt aggtttaccc gccaatatat cctgtcaaac actgatagtt
      121 taaactgaag gcgggaaacg acaatctgat ccaagctcaa gctgctctag cattcgccat
      181 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
      241 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
      301 cacgacgttg taaaacgacg gccagtgcca agcttgcatg cctgcaggtc aacatggtgg
      361 agcacgacac acttgtctac tccaaaaata tcaaagatac agtctcagaa gaccaaaggg
      421 caattgagac ttttcaacaa agggtaatat ccggaaacct cctcggattc cattgcccag
      481 ctatctgtca ctttattgtg aagatagtgg aaaaggaagg tggctcctac aaatgccatc
      541 attgcgataa aggaaaggcc atcgttgaag atgcctctgc cgacagtggt cccaaagatg
      601 gacccccacc cacgaggagc atcgtggaaa aagaagacgt tccaaccacg tcttcaaagc
      661 aagtggattg atgtgataac atggtggagc acgacacact tgtctactcc aaaaatatca
      721 aagatacagt ctcagaagac caaagggcaa ttgagacttt tcaacaaagg gtaatatccg
      781 gaaacctcct cggattccat tgcccagcta tctgtcactt tattgtgaag atagtggaaa
      841 aggaaggtgg ctcctacaaa tgccatcatt gcgataaagg aaaggccatc gttgaagatg
      901 cctctgccga cagtggtccc aaagatggac ccccacccac gaggagcatc gtggaaaaag
      961 aagacgttcc aaccacgtct tcaaagcaag tggattgatg tgatatctcc actgacgtaa
     1021 gggatgacgc acaatcccac tatccttcgc aagacccttc ctctatataa ggaagttcat
     1081 ttcatttgga gaggacctcg actctagagg atccccgggt accgggcccc ccctcgaggc
     1141 gcgccaccag gtcgacatgg ataagaagta ctccatcgga ctggatatcg gaactaactc
     1201 cgtgggatgg gctgtgatca ctgatgagta caaggtgcca tccaagaagt tcaaggtgct
     1261 gggaaacact gatagacact ccatcaagaa gaacctgatc ggagctctgc tgttcgattc
     1321 cggagagact gctgaggcta ctagactgaa gagaactgct agaagaagat acactagaag
     1381 aaagaacaga atctgctacc tgcaagagat cttctccaac gagatggcta aggtggatga
     1441 ttccttcttc cacagactgg aggagtcctt cctggtggag gaggataaga agcacgagag
     1501 acacccaatc ttcggaaaca tcgtggatga ggtggcttac cacgagaagt acccaactat
     1561 ctaccacctg agaaagaagc tggtggattc cactgataag gctgatctga gactgatcta
     1621 cctggctctg gctcacatga tcaagttcag aggacacttc ctgatcgagg gagatctgaa
     1681 cccagataac tccgatgtgg ataagctgtt catccaactg gtgcaaactt acaaccaact
     1741 gttcgaggag aacccaatca acgcttccgg agtggatgct aaggctatcc tgtccgctag
     1801 actgtccaag tccagaagac tggagaacct gatcgctcaa ctgccaggag agaagaagaa
     1861 cggactgttc ggaaacctga tcgctctgtc cctgggactg actccaaact tcaagtccaa
     1921 cttcgatctg gctgaggatg ctaagctgca actgtccaag gatacttacg atgatgatct
     1981 ggataacctg ctggctcaaa tcggagatca atacgctgat ctgttcctgg ctgctaagaa
     2041 cctgtccgat gctatcctgc tgtccgatat cctgagagtg aacactgaga tcactaaggc
     2101 tccactgtcc gcttccatga tcaagagata cgatgagcac caccaagatc tgactctgct
     2161 gaaggctctg gtgagacaac aactgccaga gaagtacaag gagatcttct tcgatcaatc
     2221 caagaacgga tacgctggat acatcgatgg aggagcttcc caagaggagt tctacaagtt
     2281 catcaagcca atcctggaga agatggatgg aactgaggag ctgctggtga agctgaacag
     2341 agaggatctg ctgagaaagc aaagaacttt cgataacgga tccatcccac accaaatcca
     2401 cctgggagag ctgcacgcta tcctgagaag acaagaggat ttctacccat tcctgaagga
     2461 taacagagag aagatcgaga agatcctgac tttcagaatc ccatactacg tgggaccact
     2521 ggctagagga aactccagat tcgcttggat gactagaaag tccgaggaga ctatcactcc
     2581 atggaacttc gaggaggtgg tggataaggg agcttccgct caatccttca tcgagagaat
     2641 gactaacttc gataagaacc tgccaaacga gaaggtgctg ccaaagcact ccctgctgta
     2701 cgagtacttc actgtgtaca acgagctgac taaggtgaag tacgtgactg agggaatgag
     2761 aaagccagct ttcctgtccg gagagcaaaa gaaggctatc gtggatctgc tgttcaagac
     2821 taacagaaag gtgactgtga agcaactgaa ggaggattac ttcaagaaga tcgagtgctt
     2881 cgattccgtg gagatctccg gagtggagga tagattcaac gcttccctgg gaacttacca
     2941 cgatctgctg aagatcatca aggataagga tttcctggat aacgaggaga acgaggatat
     3001 cctggaggat atcgtgctga ctctgactct gttcgaggat agagagatga tcgaggagag
     3061 actgaagact tacgctcacc tgttcgatga taaggtgatg aagcaactga agagaagaag
     3121 atacactgga tggggaagac tgtccagaaa gctgatcaac ggaatcagag ataagcaatc
     3181 cggaaagact atcctggatt tcctgaagtc cgatggattc gctaacagaa acttcatgca
     3241 actgatccac gatgattccc tgactttcaa ggaggatatc caaaaggctc aagtgtccgg
     3301 acaaggagat tccctgcacg agcacatcgc taacctggct ggatccccag ctatcaagaa
     3361 gggaatcctg caaactgtga aggtggtgga tgagctggtg aaggtgatgg gaagacacaa
     3421 gccagagaac atcgtgatcg agatggctag agagaaccaa actactcaaa agggacaaaa
     3481 gaactccaga gagagaatga agagaatcga ggagggaatc aaggagctgg gatcccaaat
     3541 cctgaaggag cacccagtgg agaacactca actgcaaaac gagaagctgt acctgtacta
     3601 cctgcaaaac ggaagagata tgtacgtgga tcaagagctg gatatcaaca gactgtccga
     3661 ttacgatgtg gatcacatcg tgccacaatc cttcctgaag gatgattcca tcgataacaa
     3721 ggtgctgact agatccgata agaacagagg aaagtccgat aacgtgccat ccgaggaggt
     3781 ggtgaagaag atgaagaact actggagaca actgctgaac gctaagctga tcactcaaag
     3841 aaagttcgat aacctgacta aggctgagag aggaggactg tccgagctgg ataaggctgg
     3901 attcatcaag agacaactgg tggagactag acaaatcact aagcacgtgg ctcaaatcct
     3961 ggattccaga atgaacacta agtacgatga gaacgataag ctgatcagag aggtgaaggt
     4021 gatcactctg aagtccaagc tggtgtccga tttcagaaag gatttccaat tctacaaggt
     4081 gagagagatc aacaactacc accacgctca cgatgcttac ctgaacgctg tggtgggaac
     4141 tgctctgatc aagaagtacc caaagctgga gtccgagttc gtgtacggag attacaaggt
     4201 gtacgatgtg agaaagatga tcgctaagtc cgagcaagag atcggaaagg ctactgctaa
     4261 gtacttcttc tactccaaca tcatgaactt cttcaagact gagatcactc tggctaacgg
     4321 agagatcaga aagagaccac tgatcgagac taacggagag actggagaga tcgtgtggga
     4381 taagggaaga gatttcgcta ctgtgagaaa ggtgctgtcc atgccacaag tgaacatcgt
     4441 gaagaagact gaggtgcaaa ctggaggatt ctccaaggag tccatcctgc caaagagaaa
     4501 ctccgataag ctgatcgcta gaaagaagga ttgggatcca aagaagtacg gaggattcga
     4561 ttccccaact gtggcttact ccgtgctggt ggtggctaag gtggagaagg gaaagtccaa
     4621 gaagctgaag tccgtgaagg agctgctggg aatcactatc atggagagat cctccttcga
     4681 gaagaaccca atcgatttcc tggaggctaa gggatacaag gaggtgaaga aggatctgat
     4741 catcaagctg ccaaagtact ccctgttcga gctggagaac ggaagaaaga gaatgctggc
     4801 ttccgctgga gagctgcaaa agggaaacga gctggctctg ccatccaagt acgtgaactt
     4861 cctgtacctg gcttcccact acgagaagct gaagggatcc ccagaggata acgagcaaaa
     4921 gcaactgttc gtggagcaac acaagcacta cctggatgag atcatcgagc aaatctccga
     4981 gttctccaag agagtgatcc tggctgatgc taacctggat aaggtgctgt ccgcttacaa
     5041 caagcacaga gataagccaa tcagagagca agctgagaac atcatccacc tgttcactct
     5101 gactaacctg ggagctccag ctgctttcaa gtacttcgat actactatcg atagaaagag
     5161 atacacttcc actaaggagg tgctggatgc tactctgatc caccaatcca tcactggact
     5221 gtacgagact agaatcgatc tgtcccaact gggaggagat ctgcaaccaa agaagaagag
     5281 aaaggtggga ggatgatcta gaccagatta attaactagt tctagagcgg ccgccaccgc
     5341 ggtggagctc gaatttcccc gatcgttcaa acatttggca ataaagtttc ttaagattga
     5401 atcctgttgc cggtcttgcg atgattatca tataatttct gttgaattac gttaagcatg
     5461 taataattaa catgtaatgc atgacgttat ttatgagatg ggtttttatg attagagtcc
     5521 cgcaattata catttaatac gcgatagaaa acaaaatata gcgcgcaaac taggataaat
     5581 tatcgcgcgc ggtgtcatct atgttactag atcgggaatt cgtaatcatg tcatagctgt
     5641 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     5701 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     5761 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     5821 cggggagagg cggtttgcgt attggctaga gcagcttgcc aacatggtgg agcacgacac
     5881 tctcgtctac tccaagaata tcaaagatac agtctcagaa gaccaaaggg ctattgagac
     5941 ttttcaacaa agggtaatat cgggaaacct cctcggattc cattgcccag ctatctgtca
     6001 cttcatcaaa aggacagtag aaaaggaagg tggcacctac aaatgccatc attgcgataa
     6061 aggaaaggct atcgttcaag atgcctctgc cgacagtggt cccaaagatg gacccccacc
     6121 cacgaggagc atcgtggaaa aagaagacgt tccaaccacg tcttcaaagc aagtggattg
     6181 atgtgaacat ggtggagcac gacactctcg tctactccaa gaatatcaaa gatacagtct
     6241 cagaagacca aagggctatt gagacttttc aacaaagggt aatatcggga aacctcctcg
     6301 gattccattg cccagctatc tgtcacttca tcaaaaggac agtagaaaag gaaggtggca
     6361 cctacaaatg ccatcattgc gataaaggaa aggctatcgt tcaagatgcc tctgccgaca
     6421 gtggtcccaa agatggaccc ccacccacga ggagcatcgt ggaaaaagaa gacgttccaa
     6481 ccacgtcttc aaagcaagtg gattgatgtg atatctccac tgacgtaagg gatgacgcac
     6541 aatcccacta tccttcgcaa gacccttcct ctatataagg aagttcattt catttggaga
     6601 ggacacgctg aaatcaccag tctctctcta caaatctatc tctctcgagt ctaccatgag
     6661 cccagaacga cgcccggccg acatccgccg tgccaccgag gcggacatgc cggcggtctg
     6721 caccatcgtc aaccactaca tcgagacaag cacggtcaac ttccgtaccg agccgcagga
     6781 accgcaggag tggacggacg acctcgtccg tctgcgggag cgctatccct ggctcgtcgc
     6841 cgaggtggac ggcgaggtcg ccggcatcgc ctacgcgggc ccctggaagg cacgcaacgc
     6901 ctacgactgg acggccgagt cgaccgtgta cgtctccccc cgccaccagc ggacgggact
     6961 gggctccacg ctctacaccc acctgctgaa gtccctggag gcacagggct tcaagagcgt
     7021 ggtcgctgtc atcgggctgc ccaacgaccc gagcgtgcgc atgcacgagg cgctcggata
     7081 tgccccccgc ggcatgctgc gggcggccgg cttcaagcac gggaactggc atgacgtggg
     7141 tttctggcag ctggacttca gcctgccggt accgccccgt ccggtcctgc ccgtcaccga
     7201 gatttgactc gagtttctcc ataataatgt gtgagtagtt cccagataag ggaattaggg
     7261 ttcctatagg gtttcgctca tgtgttgagc atataagaaa cccttagtat gtatttgtat
     7321 ttgtaaaata cttctatcaa taaaatttct aattcctaaa accaaaatcc agtactaaaa
     7381 tccagatccc ccgaattaat tcggcgttaa ttcagtacat taaaaacgtc cgcaatgtgt
     7441 tattaagttg tctaagcgtc aatttgttta caccacaata tatcctgcca ccagccagcc
     7501 aacagctccc cgaccggcag ctcggcacaa aatcaccact cgatacaggc agcccatcag
     7561 tccgggacgg cgtcagcggg agagccgttg taaggcggca gactttgctc atgttaccga
     7621 tgctattcgg aagaacggca actaagctgc cgggtttgaa acacggatga tctcgcggag
     7681 ggtagcatgt tgattgtaac gatgacagag cgttgctgcc tgtgatcacc gcggtttcaa
     7741 aatcggctcc gtcgatacta tgttatacgc caactttgaa aacaactttg aaaaagctgt
     7801 tttctggtat ttaaggtttt agaatgcaag gaacagtgaa ttggagttcg tcttgttata
     7861 attagcttct tggggtatct ttaaatactg tagaaaagag gaaggaaata ataaatggct
     7921 aaaatgagaa tatcaccgga attgaaaaaa ctgatcgaaa aataccgctg cgtaaaagat
     7981 acggaaggaa tgtctcctgc taaggtatat aagctggtgg gagaaaatga aaacctatat
     8041 ttaaaaatga cggacagccg gtataaaggg accacctatg atgtggaacg ggaaaaggac
     8101 atgatgctat ggctggaagg aaagctgcct gttccaaagg tcctgcactt tgaacggcat
     8161 gatggctgga gcaatctgct catgagtgag gccgatggcg tcctttgctc ggaagagtat
     8221 gaagatgaac aaagccctga aaagattatc gagctgtatg cggagtgcat caggctcttt
     8281 cactccatcg acatatcgga ttgtccctat acgaatagct tagacagccg cttagccgaa
     8341 ttggattact tactgaataa cgatctggcc gatgtggatt gcgaaaactg ggaagaagac
     8401 actccattta aagatccgcg cgagctgtat gattttttaa agacggaaaa gcccgaagag
     8461 gaacttgtct tttcccacgg cgacctggga gacagcaaca tctttgtgaa agatggcaaa
     8521 gtaagtggct ttattgatct tgggagaagc ggcagggcgg acaagtggta tgacattgcc
     8581 ttctgcgtcc ggtcgatcag ggaggatatc ggggaagaac agtatgtcga gctatttttt
     8641 gacttactgg ggatcaagcc tgattgggag aaaataaaat attatatttt actggatgaa
     8701 ttgttttagt acctagaatg catgaccaaa atcccttaac gtgagttttc gttccactga
     8761 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     8821 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     8881 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     8941 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     9001 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     9061 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     9121 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     9181 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     9241 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     9301 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     9361 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     9421 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     9481 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     9541 gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg
     9601 tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag
     9661 ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc
     9721 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     9781 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     9841 gcgcgaggca gggtgccttg atgtgggcgc cggcggtcga gtggcgacgg cgcggcttgt
     9901 ccgcgccctg gtagattgcc tggccgtagg ccagccattt ttgagcggcc agcggccgcg
     9961 ataggccgac gcgaagcggc ggggcgtagg gagcgcagcg accgaagggt aggcgctttt
    10021 tgcagctctt cggctgtgcg ctggccagac agttatgcac aggccaggcg ggttttaaga
    10081 gttttaataa gttttaaaga gttttaggcg gaaaaatcgc cttttttctc ttttatatca
    10141 gtcacttaca tgtgtgaccg gttcccaatg tacggctttg ggttcccaat gtacgggttc
    10201 cggttcccaa tgtacggctt tgggttccca atgtacgtgc tatccacagg aaagagacct
    10261 tttcgacctt tttcccctgc tagggcaatt tgccctagca tctgctccgt acattaggaa
    10321 ccggcggatg cttcgccctc gatcaggttg cggtagcgca tgactaggat cgggccagcc
    10381 tgccccgcct cctccttcaa atcgtactcc ggcaggtcat ttgacccgat cagcttgcgc
    10441 acggtgaaac agaacttctt gaactctccg gcgctgccac tgcgttcgta gatcgtcttg
    10501 aacaaccatc tggcttctgc cttgcctgcg gcgcggcgtg ccaggcggta gagaaaacgg
    10561 ccgatgccgg gatcgatcaa aaagtaatcg gggtgaaccg tcagcacgtc cgggttcttg
    10621 ccttctgtga tctcgcggta catccaatca gctagctcga tctcgatgta ctccggccgc
    10681 ccggtttcgc tctttacgat cttgtagcgg ctaatcaagg cttcaccctc ggataccgtc
    10741 accaggcggc cgttcttggc cttcttcgta cgctgcatgg caacgtgcgt ggtgtttaac
    10801 cgaatgcagg tttctaccag gtcgtctttc tgctttccgc catcggctcg ccggcagaac
    10861 ttgagtacgt ccgcaacgtg tggacggaac acgcggccgg gcttgtctcc cttcccttcc
    10921 cggtatcggt tcatggattc ggttagatgg gaaaccgcca tcagtaccag gtcgtaatcc
    10981 cacacactgg ccatgccggc cggccctgcg gaaacctcta cgtgcccgtc tggaagctcg
    11041 tagcggatca cctcgccagc tcgtcggtca cgcttcgaca gacggaaaac ggccacgtcc
    11101 atgatgctgc gactatcgcg ggtgcccacg tcatagagca tcggaacgaa aaaatctggt
    11161 tgctcgtcgc ccttgggcgg cttcctaatc gacggcgcac cggctgccgg cggttgccgg
    11221 gattctttgc ggattcgatc agcggccgct tgccacgatt caccggggcg tgcttctgcc
    11281 tcgatgcgtt gccgctgggc ggcctgcgcg gccttcaact tctccaccag gtcatcaccc
    11341 agcgccgcgc cgatttgtac cgggccggat ggtttgcgac cgctcacgcc gattcctcgg
    11401 gcttgggggt tccagtgcca ttgcagggcc ggcaggcaac ccagccgctt acgcctggcc
    11461 aaccgcccgt tcctccacac atggggcatt ccacggcgtc ggtgcctggt tgttcttgat
    11521 tttccatgcc gcctccttta gccgctaaaa ttcatctact catttattca tttgctcatt
    11581 tactctggta gctgcgcgat gtattcagat agcagctcgg taatggtctt gccttggcgt
    11641 accgcgtaca tcttcagctt ggtgtgatcc tccgccggca actgaaagtt gacccgcttc
    11701 atggctggcg tgtctgccag gctggccaac gttgcagcct tgctgctgcg tgcgctcgga
    11761 cggccggcac ttagcgtgtt tgtgcttttg ctcattttct ctttacctca ttaactcaaa
    11821 tgagttttga tttaatttca gcggccagcg cctggacctc gcgggcagcg tcgccctcgg
    11881 gttctgattc aagaacggtt gtgccggcgg cggcagtgcc tgggtagctc acgcgctgcg
    11941 tgatacggga ctcaagaatg ggcagctcgt acccggccag cgcctcggca acctcaccgc
    12001 cgatgcgcgt gcctttgatc gcccgcgaca cgacaaaggc cgcttgtagc cttccatccg
    12061 tgacctcaat gcgctgctta accagctcca ccaggtcggc ggtggcccat atgtcgtaag
    12121 ggcttggctg caccggaatc agcacgaagt cggctgcctt gatcgcggac acagccaagt
    12181 ccgccgcctg gggcgctccg tcgatcacta cgaagtcgcg ccggccgatg gccttcacgt
    12241 cgcggtcaat cgtcgggcgg tcgatgccga caacggttag cggttgatct tcccgcacgg
    12301 ccgcccaatc gcgggcactg ccctggggat cggaatcgac taacagaaca tcggccccgg
    12361 cgagttgcag ggcgcgggct agatgggttg cgatggtcgt cttgcctgac ccgcctttct
    12421 ggttaagtac agcgataacc ttcatgcgtt ccccttgcgt atttgtttat ttactcatcg
    12481 catcatatac gcagcgaccg catgacgcaa gctgttttac tcaaatacac atcacctttt
    12541 tagacggcgg cgctcggttt cttcagcggc caagctggcc ggccaggccg ccagcttggc
    12601 atcagacaaa ccggccagga tttcatgcag ccgcacggtt gagacgtgcg cgggcggctc
    12661 gaacacgtac ccggccgcga tcatctccgc ctcgatctct tcggtaatga aaaacggttc
    12721 gtcctggccg tcctggtgcg gtttcatgct tgttcctctt ggcgttcatt ctcggcggcc
    12781 gccagggcgt cggcctcggt caatgcgtcc tcacggaagg caccgcgccg cctggcctcg
    12841 gtgggcgtca cttcctcgct gcgctcaagt gcgcggtaca gggtcgagcg atgcacgcca
    12901 agcagtgcag ccgcctcttt cacggtgcgg ccttcctggt cgatcagctc gcgggcgtgc
    12961 gcgatctgtg ccggggtgag ggtagggcgg gggccaaact tcacgcctcg ggccttggcg
    13021 gcctcgcgcc cgctccgggt gcggtcgatg attagggaac gctcgaactc ggcaatgccg
    13081 gcgaacacgg tcaacaccat gcggccggcc ggcgtggtgg tgtcggccca cggctctgcc
    13141 aggctacgca ggcccgcgcc ggcctcctgg atgcgctcgg caatgtccag taggtcgcgg
    13201 gtgctgcggg ccaggcggtc tagcctggtc actgtcacaa cgtcgccagg gcgtaggtgg
    13261 tcaagcatcc tggccagctc cgggcggtcg cgcctggtgc cggtgatctt ctcggaaaac
    13321 agcttggtgc agccggccgc gtgcagttcg gcccgttggt tggtcaagtc ctggtcgtcg
    13381 gtgctgacgc gggcatagcc cagcaggcca gcggcggcgc tcttgttcat ggcgtaatgt
    13441 ctccggttct agtcgcaagt attctacttt atgcgactaa aacacgcgac aagaaaacgc
    13501 caggaaaagg gcagggcggc agcctgtcgc gtaacttagg acttgtgcga catgtcgttt
    13561 tcagaagacg gctgcactga acgtcagaag ccgactgcac tatagcagcg gaggggttgg
    13621 atcaaagtac tttgatcccg aggggaaccc tgtggttggc atg
//
