LOCUS       sequence-250173-       12096 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Gateway pENTR-plasmid to generate a binary construct for genome 
            editing of the wheat homolog of a susceptibility gene of O. sativa. 
            Encodes wheat optimized Cas9 and gene specific sgRNA modules..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12096)
  TITLE     Genome editing in wheat
REFERENCE   2  (bases 1 to 12096)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pENTRY4_ Dja6_2
FEATURES             Location/Qualifiers
     source          1..12096
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      103..189
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      281..308
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     316..339
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    358..457
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      2327..2336
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2339..2359
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      2408..2417
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2414..6517
                     /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             2414..6517
                     /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             6521..6547
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6527..6553
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     CDS             6566..6586
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6593..6613
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6629..6685
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             join(7157..7159,7160..7162,7163..7876)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     misc_RNA        8502..8577
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(8589..8613)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_RNA        9026..9101
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(9113..9137)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_RNA        9550..9625
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(9637..9661)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_RNA        10074..10149
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(10160..10184)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(10288..10307)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(10289..10307)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    complement(10325..10424)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(10536..10555)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             10547..11356
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPGDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(10614..10633)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      11446..12034
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11935..11954
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagca tggatctcgg
       61 ggacgtctaa ctactaagcg agagtaggga actgccaggc atcaaataaa acgaaaggct
      121 cagtcggagg actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
      181 aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg agggtggcgg
      241 gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat cctgacggat
      301 ggcctttttg cgtttctaca aactcttcct gttagttagt tacttaagct cgggccccaa
      361 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc
      421 ttttttataa tgccaacttt gtacaaaaaa gcaggctcca ccatggtaca aagtggtgaa
      481 gcttgcatgc ctgcagtgca gcgtgacccg gtcgtgcccc tctctagaga taatgagcat
      541 tgcatgtcta agttataaaa aattaccaca tatttttttt gtcacacttg tttgaagtgc
      601 agtttatcta tctttataca tatatttaaa ctttactcta cgaataatat aatctatagt
      661 actacaataa tatcagtgtt ttagagaatc atataaatga acagttagac atggtctaaa
      721 ggacaattga gtattttgac aacaggactc tacagtttta tctttttagt gtgcatgtgt
      781 tctccttttt ttttgcaaat agcttcacct atataatact tcatccattt tattagtaca
      841 tccatttagg gtttagggtt aatggttttt atagactaat ttttttagta catctatttt
      901 attctatttt agcctctaaa ttaagaaaac taaaactcta ttttagtttt tttatttaat
      961 aatttagata taaaatagaa taaaataaag tgactaaaaa ttaaacaaat accctttaag
     1021 aaattaaaaa aactaaggaa acatttttct tgtttcgagt agataatgcc agcctgttaa
     1081 acgccgtcga cgagtctaac ggacaccaac cagcgaacca gcagcgtcgc gtcgggccaa
     1141 gcgaagcaga cggcacggca tctctgtcgc tgcctctgga cccctctcga gagttccgct
     1201 ccaccgttgg acttgctccg ctgtcggcat ccagaaattg cgtggcggag cggcagacgt
     1261 gagccggcac ggcaggcggc ctcctcctcc tctcacggca ccggcagcta cgggggattc
     1321 ctttcccacc gctccttcgc tttcccttcc tcgcccgccg taataaatag acaccccctc
     1381 cacaccctct ttccccaacc tcgtgttgtt cggagcgcac acacacacaa ccagatctcc
     1441 cccaaatcca cccgtcggca cctccgcttc aaggtacgcc gctcgtcctc cccccccccc
     1501 cctctctacc ttctctagat cggcgttccg gtccatggtt agggcccggt agttctactt
     1561 ctgttcatgt ttgtgttaga tccgtgtttg tgttagatcc gtgctgctag cgttcgtaca
     1621 cggatgcgac ctgtacgtca gacacgttct gattgctaac ttgccagtgt ttctctttgg
     1681 ggaatcctgg gatggctcta gccgttccgc agacgggatc gatttcatga ttttttttgt
     1741 ttcgttgcat agggtttggt ttgccctttt cctttatttc aatatatgcc gtgcacttgt
     1801 ttgtcgggtc atcttttcat gctttttttt gtcttggttg tgatgatgtg gtgtggttgg
     1861 gcggtcgttc attcgttcta gatcggagta gaatactgtt tcaaactacc tggtgtattt
     1921 attaattttg gaactgtatg tgtgtgtcat acatcttcat agttacgagt ttaagatgga
     1981 tggaaatatc gatctaggat aggtatacat gttgatgtgg gttttactga tgcatataca
     2041 tgatggcata tgcagcatct attcatatgc tctaaccttg agtacctatc tattataata
     2101 aacaagtatg ttttataatt attttgatct tgatatactt ggatgatggc atatgcagca
     2161 gctatatgtg gattttttta gccctgcctt catacgctat ttatttgctt ggtactgttt
     2221 cttttgtcga tgctcaccct gttgtttggt gttacttctg caggtcgact ctagaggatc
     2281 tggaattccc ggggagaatc tggagctctc gagatttctc acgcgtgcca ccatggggcc
     2341 gaagaagaag aggaaggtgg gtagggtgtg tcggattagc agccttcggt acaggggtcc
     2401 tggcattgcc accatggaca agaagtactc tatcgggttg gatataggaa cgaatagtgt
     2461 gggttgggct gtcatcactg acgagtacaa ggtgccatcg aagaaattta aagtgctggg
     2521 caacaccgac aggcactcca tcaagaagaa cctaataggt gcgttgctct ttgattccgg
     2581 ggagacagcc gaagcgaccc gcctcaagcg cacagcaaga aggcgctata cgcgtagaaa
     2641 gaacaggatc tgctacctac aagagatctt ttccaacgaa atggcgaaag tggatgactc
     2701 cttcttccat cggctcgagg aaagcttcct cgtagaagag gacaagaagc acgaacgcca
     2761 tccgatattc gggaacattg tcgacgaagt cgcataccac gagaagtatc ctaccatcta
     2821 tcacctgagg aagaagctcg tagatagcac tgacaaggct gatttgcgcc tcatctattt
     2881 ggccctggcc catatgatta agttccgtgg gcacttcctc atcgaaggcg accttaatcc
     2941 ggacaattcc gatgttgaca aactcttcat ccaactggtc cagacataca atcagctctt
     3001 tgaagagaac cccatcaacg cgtcaggagt tgatgcgaag gcaatcttga gtgctcggct
     3061 gtcgaaaagc cggaggctcg aaaatctgat agcccaactg ccaggcgaaa agaagaacgg
     3121 attgtttggc aaccttattg cccttagtct agggctcacc cctaacttca agtctaactt
     3181 tgacctggcc gaggatgcca agctccaact gtccaaagac acgtacgacg acgatctcga
     3241 caatctgctt gcccaaatcg gcgaccagta tgcggacctt ttccttgcag cgaagaactt
     3301 gtcagatgca atcctcttgt ccgatattct gcgcgttaac actgagatta ccaaggctcc
     3361 tctttcggca tccatgatca aacggtacga cgaacatcac caggatctca cgctcctcaa
     3421 ggctctcgtc agacaacagc tgccggagaa gtacaaagag atcttcttcg atcaatcaaa
     3481 gaacgggtat gctgggtaca tcgacggtgg cgcgtcacag gaggagttct acaagtttat
     3541 caagcccatt ctcgagaaga tggatggcac tgaagagctc ctggttaagc tcaatcgcga
     3601 ggatctactc cgcaagcagc gtaccttcga caatggaagc attccacacc aaatccatct
     3661 gggcgagctt cacgccatac ttcgtaggca agaggacttt tacccattcc tcaaagacaa
     3721 ccgggagaaa atcgagaaga tcctcacatt cagaatcccc tactatgtgg gcccattggc
     3781 aaggggcaac tctaggttcg cgtggatgac tagaaaatcg gaggaaacga ttaccccatg
     3841 gaacttcgag gaggtggtgg acaagggcgc ttctgcccaa agcttcatcg agaggatgac
     3901 caacttcgac aagaatctgc caaacgagaa ggtcctcccc aaacattccc tactgtacga
     3961 gtacttcacc gtctacaacg agctgaccaa ggtcaagtac gttactgagg gtatgaggaa
     4021 accggccttt ctgtcaggag agcagaagaa ggccatagtg gatctccttt tcaagaccaa
     4081 tcgcaaggtc acagtcaagc agctaaagga ggactacttt aagaaaatcg agtgctttga
     4141 ctctgtggag ataagcggcg ttgaggatcg cttcaacgcc tcgctgggaa cgtaccacga
     4201 cttgctcaag atcattaagg ataaagactt cctcgataac gaagagaacg aggacatctt
     4261 ggaggacatc gttctgacgc ttactctctt cgaggaccgc gagatgatcg aagagaggct
     4321 caagacgtat gcgcacctct ttgacgacaa agtgatgaag cagctgaagc gcaggcgcta
     4381 cacaggttgg ggcaggctga gcaggaagct gatcaatggc attcgcgata agcagtcggg
     4441 caagaccatc ctcgacttcc tcaagtccga tgggttcgca aacaggaact ttatgcagct
     4501 aatccacgat gactccttga ccttcaagga ggacattcag aaggctcagg tttcaggcca
     4561 aggcgattcg ctccacgagc atatcgcgaa tttggccgga tctccagcca tcaagaaggg
     4621 tatacttcaa acagtgaagg ttgtcgatga gctggtcaaa gtgatgggta gacataagcc
     4681 cgagaatatc gtgattgaga tggctcggga gaaccaaacc acacagaagg ggcagaagaa
     4741 ctccagggaa aggatgaaga gaatcgaaga aggcatcaag gagctcggat cacaaattct
     4801 caaggagcat ccggtcgaaa acacccaact gcaaaacgag aagctctacc tctactacct
     4861 ccaaaacgga cgcgatatgt acgtggacca ggagctcgac atcaatcgcc tatctgacta
     4921 cgatgtcgat catatagtac cacaaagctt cctcaaagac gactctatcg acaacaaggt
     4981 gctgaccagg tccgacaaga accgcgggaa gtcagacaac gtgccttctg aggaagtcgt
     5041 caagaagatg aagaactact ggaggcaact cctcaatgca aagctcatta cacaaaggaa
     5101 gttcgacaac cttaccaagg cggagagagg cggcctcagt gagctagata aggccggatt
     5161 tatcaagcgc caactggttg agacacggca aatcaccaag cacgttgcac agatcctgga
     5221 ctcacgcatg aacacaaaat acgatgagaa tgacaagcta atccgggaag tcaaagttat
     5281 tactctgaaa tctaaactgg tgtccgactt tcggaaggac ttccagttct ataaggtccg
     5341 tgagatcaac aactaccatc acgcccatga tgcctatctc aatgctgtgg ttggcactgc
     5401 actcatcaag aaatatccca agcttgagtc ggagttcgtc tacggtgact acaaggttta
     5461 tgacgtaagg aagatgattg cgaagagtga gcaggaaatc ggcaaggcca ccgccaagta
     5521 cttcttttac agcaacataa tgaacttctt caagactgag atcaccctgg ccaatggaga
     5581 gatcagaaaa cgtccgctga tagagactaa cggagaaact ggtgagatag tgtgggacaa
     5641 ggggagagac tttgctactg tccggaaggt gctaagcatg ccacaagtca acatcgtcaa
     5701 gaaaacggaa gtccaaaccg gcggcttctc gaaagagtca atccttccga agcgcaatag
     5761 cgataagctt atagcccgta agaaagactg ggatccaaag aagtacggtg gctttgacag
     5821 tcctacagtg gcgtactccg tgctggttgt tgccaaggtg gaaaagggga agtcgaagaa
     5881 gctgaagagc gtaaaggagt tgctcgggat taccattatg gagcgttcca gctttgagaa
     5941 gaatcccatt gatttcttgg aggcgaaagg atacaaggaa gtcaagaaag atctcatcat
     6001 aaagttgccg aaatacagct tgttcgagct ggagaatggg agaaagagga tgcttgccag
     6061 cgcaggcgag ttgcaaaagg ggaacgagct ggcactccca tccaagtacg tcaacttcct
     6121 gtatctcgcc tcacactatg agaaacttaa gggttcccca gaggacaatg agcaaaagca
     6181 gctgtttgtg gagcaacaca agcactacct cgatgagatc atcgaacaga tcagtgagtt
     6241 ctcgaagagg gtgattcttg ctgacgctaa ccttgacaag gtgttgtccg cctacaacaa
     6301 gcatcgggac aaacccatcc gggagcaggc agaaaacatc atacacttgt tcacgttgac
     6361 gaaccttggc gctcctgcgg cgttcaaata ttttgacacg acgattgata ggaagcgcta
     6421 taccagcacc aaagaagtac tcgatgccac cctaattcac cagtctatta caggcctgta
     6481 cgaaacgagg atcgacttgt cgcaactagg aggagatgcc tatccgtatg acgtgccaga
     6541 ttacgcctcc cttgggtctg gttcccctaa gaagaagcgg aaagtggagg atcccaagaa
     6601 gaagcgcaaa gtcgacggcg gatccggagc cacgaacttt tccctgctca agcaagctgg
     6661 tgatgtggag gagaatcctg gcccagctcc aggctcaacg tccatggcga aggcagataa
     6721 gaaaccagcg gagaagaaac cggcagagaa aactccggca gtcgaaccag cagcagcggc
     6781 agagaagaaa ccaaaagccg gaaagaaact ccccaaggaa ccagccgtcg ccggagacaa
     6841 gaagaagaag agatcaaaga agaacgttga gacatacaag atctacatct tcaaggtgtt
     6901 gaagcaagtt catccagaca tcggaatctc aagcaaagcg atgggaatta tgaacagttt
     6961 catcaacgat atttttgaga agctcgctgg tgaatcttcg aagctggcga ggtacaacaa
     7021 gaagccgacg attacttcca gggagattca gactgcggtt agacttgtgt tgcccggtga
     7081 gttggcgaaa catgctgtgt ctgaagggac taaggcggtt acgaagttta cgagttctgc
     7141 tcagggcgcc tctttcatgg tgtccaaagg agaggagctc tttacaggag ttgtgcccat
     7201 tttggtcgag ttggatggcg acgtaaatgg ccacaagttc agcgtgagag gcgaaggtga
     7261 aggtgatgcc accaatggca agcttaccct caagttcatc tgtaccactg ggaaactgcc
     7321 ggttccatgg ccaacgttgg tcaccactct cacatatggt gttcagtgct tttcgaggta
     7381 tccggaccac atgaaacgcc atgacttctt caagtcagcc atgcctgagg ggtatgtcca
     7441 agaacgcacc atcagcttta aggacgacgg aacgtacaag acacgtgcgg aggtcaagtt
     7501 cgagggagat acactggtga acaggatcga actcaaaggg atcgacttca aagaggacgg
     7561 caacatactg gggcataagc tcgagtacaa cttcaattct cacaacgtct acattactgc
     7621 cgacaagcag aagaacggca ttaaggcgaa cttcaaaatc cggcataatg tggaggatgg
     7681 tagtgtccaa ctggctgacc actaccagca gaacacgccg ataggcgatg gcccagtgct
     7741 tctgcccgac aatcactacc tgtcaaccca atccgttctg tctaaggacc ctaacgaaaa
     7801 gagggatcac atggtgcttc tggagtttgt aacggctgca ggcatcactc atgggatgga
     7861 tgagctctac aagtgataac tgcggccgca gatatcacta gtgttcaaac atttggcaat
     7921 aaagtttctt aagattgaat cctgttgccg gtcttgcgat gattatcata taatttctgt
     7981 tgaattacgt taagcatgta ataattaaca tgtaatgcat gacgttattt atgagatggg
     8041 tttttatgat tagagtcccg caattataca tttaatacgc gatagaaaac aaaatataac
     8101 tagaattcga gctcggagga ccaagcccgt tattctgaca gttctggtgc tcaacacatt
     8161 tatatttatc aaggagcaca ttgttactca ctgctaggag ggaatcgaac taggaatatt
     8221 gatcagagga actacgagag agctgaagat aactgccctc tagctctcac tgatctgggt
     8281 cgcatagtga gatgcagccc acgtgagttc agcaacggtc tagcgctggg cttttaggcc
     8341 cgcatgatcg ggcttttgtc gggtggtcga cgtgttcacg attggggaga gcaacgcagc
     8401 agttcctctt agtttagtcc cacctcgcct gtccagcaga gttctgaccg gtttataaac
     8461 tcgcttgctg catcagactt ggtgttgttg ctggtcttcc tgttttagag ctagaaatag
     8521 caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt
     8581 ttttctagac ccagctttct tgtacaaagt tggcattacg ctttacgaat tcccatgggg
     8641 aggaccaagc ccgttattct gacagttctg gtgctcaaca catttatatt tatcaaggag
     8701 cacattgtta ctcactgcta ggagggaatc gaactaggaa tattgatcag aggaactacg
     8761 agagagctga agataactgc cctctagctc tcactgatct gggtcgcata gtgagatgca
     8821 gcccacgtga gttcagcaac ggtctagcgc tgggctttta ggcccgcatg atcgggcttt
     8881 tgtcgggtgg tcgacgtgtt cacgattggg gagagcaacg cagcagttcc tcttagttta
     8941 gtcccacctc gcctgtccag cagagttctg accggtttat aaactcgctt gctgcatcag
     9001 acttggtgct gtcgctcttc ttcgggtttt agagctagaa atagcaagtt aaaataaggc
     9061 tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg ctttttttct agacccagct
     9121 ttcttgtaca aagttggcat tacgctcaga gaattcgcat gcggaggacc aagcccgtta
     9181 ttctgacagt tctggtgctc aacacattta tatttatcaa ggagcacatt gttactcact
     9241 gctaggaggg aatcgaacta ggaatattga tcagaggaac tacgagagag ctgaagataa
     9301 ctgccctcta gctctcactg atctgggtcg catagtgaga tgcagcccac gtgagttcag
     9361 caacggtcta gcgctgggct tttaggcccg catgatcggg cttttgtcgg gtggtcgacg
     9421 tgttcacgat tggggagagc aacgcagcag ttcctcttag tttagtccca cctcgcctgt
     9481 ccagcagagt tctgaccggt ttataaactc gcttgctgca tcagacttgt tcgaacagct
     9541 atttgggggg ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact
     9601 tgaaaaagtg gcaccgagtc ggtgcttttt ttctagaccc agctttcttg tacaaagttg
     9661 gcattacgct tgtggaattc ctcgagggag gaccaagccc gttattctga cagttctggt
     9721 gctcaacaca tttatattta tcaaggagca cattgttact cactgctagg agggaatcga
     9781 actaggaata ttgatcagag gaactacgag agagctgaag ataactgccc tctagctctc
     9841 actgatctgg gtcgcatagt gagatgcagc ccacgtgagt tcagcaacgg tctagcgctg
     9901 ggcttttagg cccgcatgat cgggcttttg tcgggtggtc gacgtgttca cgattgggga
     9961 gagcaacgca gcagttcctc ttagtttagt cccacctcgc ctgtccagca gagttctgac
    10021 cggtttataa actcgcttgc tgcatcagac ttgcatcaag aaccaccccg acagttttag
    10081 agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg
    10141 agtcggtgct tttttctaga cccagctttc ttgtacaaag ttggcattac gctgagcgag
    10201 gatgcacatg tgaccgaggg aggtgtcatc tatgttactg ctagtcgatt taaatcctta
    10261 attaaccctg caggcatctc caattcgccc tatagtgagt cgtattacac cggtgaggat
    10321 ctagacccag ctttcttgta caaagttggc attataagaa agcattgctt atcaatttgt
    10381 tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc agctgcagct
    10441 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
    10501 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
    10561 acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa
    10621 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc
    10681 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
    10741 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
    10801 tatccgtact cctggtgatg catggttact caccactgcg atccccggaa aaacagcatt
    10861 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
    10921 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
    10981 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
    11041 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc
    11101 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
    11161 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
    11221 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
    11281 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
    11341 cgatgagttt ttctaatcag aattggttaa ttggttgtaa cattattcag attgggcccc
    11401 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
    11461 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
    11521 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
    11581 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
    11641 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
    11701 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
    11761 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
    11821 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
    11881 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
    11941 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
    12001 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
    12061 gttcctggcc ttttgctggc cttttgctca catgtt
//
