LOCUS       sequence-249713-       17296 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genome editing of the wheat homolog of a susceptibility gene of O. 
            sativa. Encodes wheat optimized Cas9 module, nptII resistance gene, 
            and specific sgRNA modules on the binary plasmid pICSL4723..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 17296)
  TITLE     Genome editing in wheat
REFERENCE   2  (bases 1 to 17296)
  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"
COMMENT     Sequence Label: pICSL4723_ Rac4
FEATURES             Location/Qualifiers
     source          1..17296
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    32..56
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             365..994
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             1423..2496
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGASSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      2562..2756
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     promoter        2818..2922
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2923..3717
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      3888..4476
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4377..4396
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4612..4636
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(4727..4979)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(5103..5122)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6017..6036
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(6633..6650)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        complement(6653..7495)
                     /label=Act1 promoter
                     /note="rice actin promoter"
     CDS             complement(join(7751..8464,8465..8467,8468..8470))
                     /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"
     CDS             complement(8942..8998)
                     /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             complement(9014..9034)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(9041..9061)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(9074..9100)
                     /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             complement(9080..9106)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(9110..13213)
                     /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             complement(9110..13213)
                     /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"
     regulatory      13210..13219
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(13268..13288)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      13291..13300
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_RNA        15576..15651
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(15663..15687)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_RNA        16100..16175
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(16187..16211)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_RNA        16624..16699
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(16711..16735)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_RNA        17148..17223
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(17235..17259)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 cacgaagtga tccgtttaaa ctatcagtgt ttgacaggat atattggcgg gtaaacctaa
       61 gagaaaagag cgtttattag aataatcgga tatttaaaag ggcgtgaaaa ggtttatccg
      121 ttcgtccatt tgtatgtgca tgccaaccac agggttcccc tcgggagtca gccgtgcggc
      181 tgcatgaaat cctggccggt ttgtctgatg ccaagctggc ggcctggccg gccagcttgg
      241 ccgctgaaga aaccgagcgc cgccgtctaa aaaggtgatg tgtatttgag taaaacagct
      301 tgcgtcatgc ggtcgctgcg tatatgatgc gatgagtaaa taaacaaata cgcaagggga
      361 acgcatgaag gttatcgctg tacttaacca gaaaggcggg tcaggcaaga cgaccatcgc
      421 aacccatcta gcccgcgccc tgcaactcgc cggggccgat gttctgttag tcgattccga
      481 tccccagggc agtgcccgcg attgggcggc cgtgcgggaa gatcaaccgc taaccgttgt
      541 cggcatcgac cgcccgacga ttgaccgcga cgtgaaggcc atcggccggc gcgacttcgt
      601 agtgatcgac ggagcgcccc aggcggcgga cttggctgtg tccgcgatca aggcagccga
      661 cttcgtgctg attccggtgc agccaagccc ttacgacata tgggccaccg ccgacctggt
      721 ggagctggtt aagcagcgca ttgaggtcac ggatggaagg ctacaagcgg cctttgtcgt
      781 gtcgcgggcg atcaaaggca cgcgcatcgg cggtgaggtt gccgaggcgc tggccgggta
      841 cgagctgccc attcttgagt cccgtatcac gcagcgcgtg agctacccag gcactgccgc
      901 cgccggcaca accgttcttg aatcagaacc cgagggcgac gctgcccgcg aggtccaggc
      961 gctggccgct gaaattaaat caaaactcat ttgagttaat gaggtaaaga gaaaatgagc
     1021 aaaagcacaa acacgctaag tgccggccgt ccgagcgcac gcagcagcaa ggctgcaacg
     1081 ttggccagcc tggcagacac gccagccatg aagcgggtca actttcagtt gccggcggag
     1141 gatcacacca agctgaagat gtacgcggta cgccaaggca agaccattac cgagctgcta
     1201 tctgaataca tcgcgcagct accagagtaa atgagcaaat gaataaatga gtagatgaat
     1261 tttagcggct aaaggaggcg gcatggaaaa tcaagaacaa ccaggcaccg acgccgtgga
     1321 atgccccatg tgtggaggaa cgggcggttg gccaggcgta agcggctggg ttgtctgccg
     1381 gccctgcaat ggcactggaa cccccaagcc cgaggaatcg gcgtgagcgg tcgcaaacca
     1441 tccggcccgg tacaaatcgg cgcggcgctg ggtgatgacc tggtggagaa gttgaaggcc
     1501 gcgcaggccg cccagcggca acgcatcgag gcagaagcac gccccggtga atcgtggcaa
     1561 gcggccgctg atcgaatccg caaagaatcc cggcaaccgc cggcagccgg tgcgtcgtcg
     1621 attaggaagc cgcccaaggg cgacgagcaa ccagattttt tcgttccgat gctctatgac
     1681 gtgggcaccc gcgatagtcg cagcatcatg gacgtggccg ttttccgtct gtcgaagcgt
     1741 gaccgacgag ctggcgaggt gatccgctac gagcttccag acgggcacgt agaggtttcc
     1801 gcagggccgg ccggcatggc cagtgtgtgg gattacgacc tggtactgat ggcggtttcc
     1861 catctaaccg aatccatgaa ccgataccgg gaagggaagg gagacaagcc cggccgcgtg
     1921 ttccgtccac acgttgcgga cgtactcaag ttctgccggc gagccgatgg cggaaagcag
     1981 aaagacgacc tggtagaaac ctgcattcgg ttaaacacca cgcacgttgc catgcagcgt
     2041 acgaagaagg ccaagaacgg ccgcctggtg acggtatccg agggtgaagc cttgattagc
     2101 cgctacaaga tcgtaaagag cgaaaccggg cggccggagt acatcgagat cgagctagct
     2161 gattggatgt accgcgagat tacagaaggc aagaacccgg acgtgctgac ggttcacccc
     2221 gattactttt tgatcgatcc cggcatcggc cgttttctct accgcctggc acgccgcgcc
     2281 gcaggcaagg cagaagccag atggttgttc aagacgatct acgaacgcag tggcagcgcc
     2341 ggagagttca agaagttctg tttcaccgtg cgcaagctga tcgggtcaaa tgacctgccg
     2401 gagtacgatt tgaaggagga ggcggggcag gctggcccga tcctagtcat gcgctaccgc
     2461 aacctgatcg agggcgaagc atccgccggt tcctaatgta cggagcagat gctagggcaa
     2521 attgccctag caggggaaaa aggtcgaaaa agcttctttc ctgtggatag cacgtacatt
     2581 gggaacccaa agccgtacat tgggaaccgg aacccgtaca ttgggaaccc aaagccgtac
     2641 attgggaacc ggtcacacat gtaagtgact gatataaaag agaaaaaagg cgatttttcc
     2701 gcctaaaact ctttaaaact tattaaaact cttaaaaccc gcctggcctg tgcataactg
     2761 tctggccagc gcacagccga acagctgcaa aaagcgccca catcaaggct ccgagtgcgc
     2821 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     2881 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatggctaa aatgagaata
     2941 tcaccggaat tgaaaaaact gatcgaaaaa taccgctgcg taaaagatac ggaaggaatg
     3001 tctcctgcta aggtatataa gctggtggga gaaaatgaaa acctatattt aaaaatgacg
     3061 gacagccggt ataaagggac cacctatgat gtggaacggg aaaaggacat gatgctatgg
     3121 ctggaaggaa agctgcctgt tccaaaggtc ctgcactttg aacggcatga tggctggagc
     3181 aatctgctca tgagtgaggc cgatggcgtc ctttgctcgg aagagtatga agatgaacaa
     3241 agccctgaaa agattatcga gctgtatgcg gagtgcatca ggctctttca ctccatcgac
     3301 atatcggatt gtccctatac gaatagctta gacagccgct tagccgaatt ggattactta
     3361 ctgaataacg atctggccga tgtggattgc gaaaactggg aagaggacac tccatttaaa
     3421 gatccgcgcg agctgtatga ttttttaaag acggaaaagc ccgaagagga acttgtcttt
     3481 tcccacggcg acctgggaga cagcaacatc tttgtgaaag atggcaaagt aagtggcttt
     3541 attgatcttg ggagaagcgg cagggcggac aagtggtatg acattgcctt ctgcgtccgg
     3601 tcgctcaggg aggatatcgg ggaagaacag tatgtcgagc tattttttga cttactgggg
     3661 atcaagcctg attgggagaa aataaaatat tatattttac tggatgaatt gttttagctg
     3721 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     3781 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     3841 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     3901 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     3961 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     4021 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     4081 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     4141 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     4201 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     4261 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     4321 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     4381 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     4441 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     4501 cggttcctgc tcggatctgt tggaccggac agtagtcatg gttgatgggc tgcctgtatc
     4561 gagtggtgat tttgtgccga gctgccggtc ggggagctgt tggctggctg gtggcaggat
     4621 atattgtggt gtaaacaaat tgacgcttag acaacttaat aacacattgc ggacgttttt
     4681 aatgtactgg ggttgaacac tctgtgccga attcggatcc agcgtcgatc tagtaacata
     4741 gatgacaccg cgcgcgataa tttatcctag tttgcgcgct atattttgtt ttctatcgcg
     4801 tattaaatgt ataattgcgg gactctaatc ataaaaaccc atctcataaa taacgtcatg
     4861 cattacatgt taattattac atgcttaacg taattcaaca gaaattatat gataatcatc
     4921 gcaagaccgg caacaggatt caatcttaag aaactttatt gccaaatgtt tgaacgatct
     4981 gcttgacaag ctcagaagaa ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg
     5041 ggagcggcga taccgtaaag cacgaggaag cggtcagccc attcgccgcc aagctcttca
     5101 gcaatatcac gggtagccaa cgctatgtcc tgatagcggt ccgccacacc cagccggcca
     5161 cagtcgatga atccagaaaa gcggccattt tccaccatga tattcggcaa gcaggcatcg
     5221 ccgtgggtca cgacgagatc ctcgccgtcg ggcatccgcg ccttgagcct ggcgaacagt
     5281 tctgttaatc agaaaaactc ggattaatcg acaaattcga tcgcacaaac tagaaactaa
     5341 caccagatct agatagaaat cacaagtcga agagtaatta ttcgacaaaa ctcaaattat
     5401 ttgaacaaat cggatgatat ctatgaaacc ctaatcgaga attaagatga tatctaacga
     5461 tcaaacccag aaaatcgtct tagatctaag attaacagaa tctaaaccaa agaacatata
     5521 cgaaattggg atcgaacgaa aacaaaatcg aagattttga gagaataagg aacacagaaa
     5581 cttacctgca ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat cgacaagacc
     5641 ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca
     5701 ggtagccgga tcaagcgtat gcagccgccg cattgcatca gccatgatgg atactttctc
     5761 ggcaggagca aggtgagatg acaggagatc ctgccccggc acttcgccca atagcagcca
     5821 gtcccttccc gcttcagtga caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc
     5881 cagccacgat agccgcgctg cctcgtcttg gagttcattc agggcaccgg acaggtcggt
     5941 cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg aacacggcgg catcagagca
     6001 gccgattgtc tgttgtgccc agtcatagcc gaatagcctc tccacccaag cggccggaga
     6061 acctgcgtgc aatccatctt gttcaatcat tggcttctac ctacaaaaaa gctccgcacg
     6121 aggctgcatt tgtcacaaat catgaaaaga aaaactaccg atgaacaatg ctgagggatt
     6181 caaattctac ccacaaaaag aagaaagaaa gatctagcac atctaagcct gacgaagcag
     6241 cagaaatata taaaaatata aaccatagtg cccttttccc ctcttcctga tcttgtttag
     6301 catggcggaa attttaaacc ccccatcatc tcccccaaca acggcggatc gcagatctac
     6361 atccgagagc cccattcccc gcgagatccg ggccggatcc acgccggcga gagccccagc
     6421 cgcgagatcc cgcccctccc gcgcaccgat ctgggcgcgc acgaagccgc ctctcgccca
     6481 cccaaactac caaggccaaa gatcgtgtcc gagacggaaa aaaaaaacgg agaaagaaag
     6541 aggagagggg cggggtggtt accggcgcgg cggcggcgga gggggagggg ggaggagctc
     6601 gtcgtccggc agcgaggggg gaggaggtgg aggtggtggt ggtggtggtg gtagggttgg
     6661 ggggatggga ggagaggggg gggtatgtat atagtggcga tggggggcgt ttctttggaa
     6721 gcggagggag ggccggcctc gtcgctggct cgcgatcctc ctcgcgtttc cggcccccac
     6781 gacccggacc cacctgctgt tttttctttt tctttttttt ctttcttttt ttttttttgg
     6841 ctgcgagacg tgcggtgcgt gcggacaact cacggtgata gtgggggggt gtggagacta
     6901 ttgtccagtt ggctggactg gggtgggttg ggttgggttg ggttgggctg ggcttgctat
     6961 ggatcgtgga tagcactttg ggctttagga actttagggg ttgtttttgt aaatgttttg
     7021 agtctaagtt tatcttttat ttttactaga aaaaataccc atgcgctgca acgggggaaa
     7081 gctattttaa tcttattatt gttcattgtg agaattcgcc tgaatatata tttttctcaa
     7141 aaattatgtc aaattagcat atgggttttt ttaaagatat ttcttataca aatccctctg
     7201 tatttacaaa agcaaacgaa cttaaaaccc gactcaaata cagatatgca tttccaaaag
     7261 cgaataaact taaaaaccaa ttcatacaaa aatgacgtat caaagtaccg acaaaaacat
     7321 cctcaatttt tataatagta gaaaagagta aatttcactt tgggccacct tttattaccg
     7381 atattttact ttataccacc ttttaactga tgttttcact tttgaccagg taatcttacc
     7441 tttgttttat tttggactat cccgactctc ttctcaagca tatgaatgac ctcgagtatg
     7501 ctagctccgc aagaattcaa gcttagcgta tattttgttt tctatcgcgt attaaatgta
     7561 taattgcggg actctaatca taaaaaccca tctcataaat aacgtcatgc attacatgtt
     7621 aattattaca tgcttaacgt aattcaacag aaattatatg ataatcatcg caagaccggc
     7681 aacaggattc aatcttaaga aactttattg ccaaatgttt gaacactagt gatatctgcg
     7741 gccgcagtta tcacttgtag agctcatcca tcccatgagt gatgcctgca gccgttacaa
     7801 actccagaag caccatgtga tccctctttt cgttagggtc cttagacaga acggattggg
     7861 ttgacaggta gtgattgtcg ggcagaagca ctgggccatc gcctatcggc gtgttctgct
     7921 ggtagtggtc agccagttgg acactaccat cctccacatt atgccggatt ttgaagttcg
     7981 ccttaatgcc gttcttctgc ttgtcggcag taatgtagac gttgtgagaa ttgaagttgt
     8041 actcgagctt atgccccagt atgttgccgt cctctttgaa gtcgatccct ttgagttcga
     8101 tcctgttcac cagtgtatct ccctcgaact tgacctccgc acgtgtcttg tacgttccgt
     8161 cgtccttaaa gctgatggtg cgttcttgga catacccctc aggcatggct gacttgaaga
     8221 agtcatggcg tttcatgtgg tccggatacc tcgaaaagca ctgaacacca tatgtgagag
     8281 tggtgaccaa cgttggccat ggaaccggca gtttcccagt ggtacagatg aacttgaggg
     8341 taagcttgcc attggtggca tcaccttcac cttcgcctct cacgctgaac ttgtggccat
     8401 ttacgtcgcc atccaactcg accaaaatgg gcacaactcc tgtaaagagc tcctctcctt
     8461 tggacaccat gaaagaggcg ccctgagcag aactcgtaaa cttcgtaacc gccttagtcc
     8521 cttcagacac agcatgtttc gccaactcac cgggcaacac aagtctaacc gcagtctgaa
     8581 tctccctgga agtaatcgtc ggcttcttgt tgtacctcgc cagcttcgaa gattcaccag
     8641 cgagcttctc aaaaatatcg ttgatgaaac tgttcataat tcccatcgct ttgcttgaga
     8701 ttccgatgtc tggatgaact tgcttcaaca ccttgaagat gtagatcttg tatgtctcaa
     8761 cgttcttctt tgatctcttc ttcttcttgt ctccggcgac ggctggttcc ttggggagtt
     8821 tctttccggc ttttggtttc ttctctgccg ctgctgctgg ttcgactgcc ggagttttct
     8881 ctgccggttt cttctccgct ggtttcttat ctgccttcgc catggacgtt gagcctggag
     8941 ctgggccagg attctcctcc acatcaccag cttgcttgag cagggaaaag ttcgtggctc
     9001 cggatccgcc gtcgactttg cgcttcttct tgggatcctc cactttccgc ttcttcttag
     9061 gggaaccaga cccaagggag gcgtaatctg gcacgtcata cggataggca tctcctccta
     9121 gttgcgacaa gtcgatcctc gtttcgtaca ggcctgtaat agactggtga attagggtgg
     9181 catcgagtac ttctttggtg ctggtatagc gcttcctatc aatcgtcgtg tcaaaatatt
     9241 tgaacgccgc aggagcgcca aggttcgtca acgtgaacaa gtgtatgatg ttttctgcct
     9301 gctcccggat gggtttgtcc cgatgcttgt tgtaggcgga caacaccttg tcaaggttag
     9361 cgtcagcaag aatcaccctc ttcgagaact cactgatctg ttcgatgatc tcatcgaggt
     9421 agtgcttgtg ttgctccaca aacagctgct tttgctcatt gtcctctggg gaacccttaa
     9481 gtttctcata gtgtgaggcg agatacagga agttgacgta cttggatggg agtgccagct
     9541 cgttcccctt ttgcaactcg cctgcgctgg caagcatcct ctttctccca ttctccagct
     9601 cgaacaagct gtatttcggc aactttatga tgagatcttt cttgacttcc ttgtatcctt
     9661 tcgcctccaa gaaatcaatg ggattcttct caaagctgga acgctccata atggtaatcc
     9721 cgagcaactc ctttacgctc ttcagcttct tcgacttccc cttttccacc ttggcaacaa
     9781 ccagcacgga gtacgccact gtaggactgt caaagccacc gtacttcttt ggatcccagt
     9841 ctttcttacg ggctataagc ttatcgctat tgcgcttcgg aaggattgac tctttcgaga
     9901 agccgccggt ttggacttcc gttttcttga cgatgttgac ttgtggcatg cttagcacct
     9961 tccggacagt agcaaagtct ctccccttgt cccacactat ctcaccagtt tctccgttag
    10021 tctctatcag cggacgtttt ctgatctctc cattggccag ggtgatctca gtcttgaaga
    10081 agttcattat gttgctgtaa aagaagtact tggcggtggc cttgccgatt tcctgctcac
    10141 tcttcgcaat catcttcctt acgtcataaa ccttgtagtc accgtagacg aactccgact
    10201 caagcttggg atatttcttg atgagtgcag tgccaaccac agcattgaga taggcatcat
    10261 gggcgtgatg gtagttgttg atctcacgga ccttatagaa ctggaagtcc ttccgaaagt
    10321 cggacaccag tttagatttc agagtaataa ctttgacttc ccggattagc ttgtcattct
    10381 catcgtattt tgtgttcatg cgtgagtcca ggatctgtgc aacgtgcttg gtgatttgcc
    10441 gtgtctcaac cagttggcgc ttgataaatc cggccttatc tagctcactg aggccgcctc
    10501 tctccgcctt ggtaaggttg tcgaacttcc tttgtgtaat gagctttgca ttgaggagtt
    10561 gcctccagta gttcttcatc ttcttgacga cttcctcaga aggcacgttg tctgacttcc
    10621 cgcggttctt gtcggacctg gtcagcacct tgttgtcgat agagtcgtct ttgaggaagc
    10681 tttgtggtac tatatgatcg acatcgtagt cagataggcg attgatgtcg agctcctggt
    10741 ccacgtacat atcgcgtccg ttttggaggt agtagaggta gagcttctcg ttttgcagtt
    10801 gggtgttttc gaccggatgc tccttgagaa tttgtgatcc gagctccttg atgccttctt
    10861 cgattctctt catcctttcc ctggagttct tctgcccctt ctgtgtggtt tggttctccc
    10921 gagccatctc aatcacgata ttctcgggct tatgtctacc catcactttg accagctcat
    10981 cgacaacctt cactgtttga agtataccct tcttgatggc tggagatccg gccaaattcg
    11041 cgatatgctc gtggagcgaa tcgccttggc ctgaaacctg agccttctga atgtcctcct
    11101 tgaaggtcaa ggagtcatcg tggattagct gcataaagtt cctgtttgcg aacccatcgg
    11161 acttgaggaa gtcgaggatg gtcttgcccg actgcttatc gcgaatgcca ttgatcagct
    11221 tcctgctcag cctgccccaa cctgtgtagc gcctgcgctt cagctgcttc atcactttgt
    11281 cgtcaaagag gtgcgcatac gtcttgagcc tctcttcgat catctcgcgg tcctcgaaga
    11341 gagtaagcgt cagaacgatg tcctccaaga tgtcctcgtt ctcttcgtta tcgaggaagt
    11401 ctttatcctt aatgatcttg agcaagtcgt ggtacgttcc cagcgaggcg ttgaagcgat
    11461 cctcaacgcc gcttatctcc acagagtcaa agcactcgat tttcttaaag tagtcctcct
    11521 ttagctgctt gactgtgacc ttgcgattgg tcttgaaaag gagatccact atggccttct
    11581 tctgctctcc tgacagaaag gccggtttcc tcataccctc agtaacgtac ttgaccttgg
    11641 tcagctcgtt gtagacggtg aagtactcgt acagtaggga atgtttgggg aggaccttct
    11701 cgtttggcag attcttgtcg aagttggtca tcctctcgat gaagctttgg gcagaagcgc
    11761 ccttgtccac cacctcctcg aagttccatg gggtaatcgt ttcctccgat tttctagtca
    11821 tccacgcgaa cctagagttg ccccttgcca atgggcccac atagtagggg attctgaatg
    11881 tgaggatctt ctcgattttc tcccggttgt ctttgaggaa tgggtaaaag tcctcttgcc
    11941 tacgaagtat ggcgtgaagc tcgcccagat ggatttggtg tggaatgctt ccattgtcga
    12001 aggtacgctg cttgcggagt agatcctcgc gattgagctt aaccaggagc tcttcagtgc
    12061 catccatctt ctcgagaatg ggcttgataa acttgtagaa ctcctcctgt gacgcgccac
    12121 cgtcgatgta cccagcatac ccgttctttg attgatcgaa gaagatctct ttgtacttct
    12181 ccggcagctg ttgtctgacg agagccttga ggagcgtgag atcctggtga tgttcgtcgt
    12241 accgtttgat catggatgcc gaaagaggag ccttggtaat ctcagtgtta acgcgcagaa
    12301 tatcggacaa gaggattgca tctgacaagt tcttcgctgc aaggaaaagg tccgcatact
    12361 ggtcgccgat ttgggcaagc agattgtcga gatcgtcgtc gtacgtgtct ttggacagtt
    12421 ggagcttggc atcctcggcc aggtcaaagt tagacttgaa gttaggggtg agccctagac
    12481 taagggcaat aaggttgcca aacaatccgt tcttcttttc gcctggcagt tgggctatca
    12541 gattttcgag cctccggctt ttcgacagcc gagcactcaa gattgccttc gcatcaactc
    12601 ctgacgcgtt gatggggttc tcttcaaaga gctgattgta tgtctggacc agttggatga
    12661 agagtttgtc aacatcggaa ttgtccggat taaggtcgcc ttcgatgagg aagtgcccac
    12721 ggaacttaat catatgggcc agggccaaat agatgaggcg caaatcagcc ttgtcagtgc
    12781 tatctacgag cttcttcctc aggtgataga tggtaggata cttctcgtgg tatgcgactt
    12841 cgtcgacaat gttcccgaat atcggatggc gttcgtgctt cttgtcctct tctacgagga
    12901 agctttcctc gagccgatgg aagaaggagt catccacttt cgccatttcg ttggaaaaga
    12961 tctcttgtag gtagcagatc ctgttctttc tacgcgtata gcgccttctt gctgtgcgct
    13021 tgaggcgggt cgcttcggct gtctccccgg aatcaaagag caacgcacct attaggttct
    13081 tcttgatgga gtgcctgtcg gtgttgccca gcactttaaa tttcttcgat ggcaccttgt
    13141 actcgtcagt gatgacagcc caacccacac tattcgttcc tatatccaac ccgatagagt
    13201 acttcttgtc catggtggca atgccaggac ccctgtaccg aaggctgcta atccgacaca
    13261 ccctacccac cttcctcttc ttcttcggcc ccatggtggc acgcgtgaga aatctcgaga
    13321 gctccagatt ctccccggga attccagatc ctctagagtc gacctgcaga agtaacacca
    13381 aacaacaggg tgagcatcga caaaagaaac agtaccaagc aaataaatag cgtatgaagg
    13441 cagggctaaa aaaatccaca tatagctgct gcatatgcca tcatccaagt atatcaagat
    13501 caaaataatt ataaaacata cttgtttatt ataatagata ggtactcaag gttagagcat
    13561 atgaatagat gctgcatatg ccatcatgta tatgcatcag taaaacccac atcaacatgt
    13621 atacctatcc tagatcgata tttccatcca tcttaaactc gtaactatga agatgtatga
    13681 cacacacata cagttccaaa attaataaat acaccaggta gtttgaaaca gtattctact
    13741 ccgatctaga acgaatgaac gaccgcccaa ccacaccaca tcatcacaac caagacaaaa
    13801 aaaagcatga aaagatgacc cgacaaacaa gtgcacggca tatattgaaa taaaggaaaa
    13861 gggcaaacca aaccctatgc aacgaaacaa aaaaaatcat gaaatcgatc ccgtctgcgg
    13921 aacggctaga gccatcccag gattccccaa agagaaacac tggcaagtta gcaatcagaa
    13981 cgtgtctgac gtacaggtcg catccgtgta cgaacgctag cagcacggat ctaacacaaa
    14041 cacggatcta acacaaacat gaacagaagt agaactaccg ggccctaacc atggaccgga
    14101 acgccgatct agagaaggta gagagggggg gggggggagg acgagcggcg taccttgaag
    14161 cggaggtgcc gacgggtgga tttgggggag atctggttgt gtgtgtgtgc gctccgaaca
    14221 acacgaggtt ggggaaagag ggtgtggagg gggtgtctat ttattacggc gggcgaggaa
    14281 gggaaagcga aggagcggtg ggaaaggaat cccccgtagc tgccggtgcc gtgagaggag
    14341 gaggaggccg cctgccgtgc cggctcacgt ctgccgctcc gccacgcaat ttctggatgc
    14401 cgacagcgga gcaagtccaa cggtggagcg gaactctcga gaggggtcca gaggcagcga
    14461 cagagatgcc gtgccgtctg cttcgcttgg cccgacgcga cgctgctggt tcgctggttg
    14521 gtgtccgtta gactcgtcga cggcgtttaa caggctggca ttatctactc gaaacaagaa
    14581 aaatgtttcc ttagtttttt taatttctta aagggtattt gtttaatttt tagtcacttt
    14641 attttattct attttatatc taaattatta aataaaaaaa ctaaaataga gttttagttt
    14701 tcttaattta gaggctaaaa tagaataaaa tagatgtact aaaaaaatta gtctataaaa
    14761 accattaacc ctaaacccta aatggatgta ctaataaaat ggatgaagta ttatataggt
    14821 gaagctattt gcaaaaaaaa aggagaacac atgcacacta aaaagataaa actgtagagt
    14881 cctgttgtca aaatactcaa ttgtccttta gaccatgtct aactgttcat ttatatgatt
    14941 ctctaaaaca ctgatattat tgtagtacta tagattatat tattcgtaga gtaaagttta
    15001 aatatatgta taaagataga taaactgcac ttcaaacaag tgtgacaaaa aaaatatgtg
    15061 gtaatttttt ataacttaga catgcaatgc tcattatctc tagagagggg cacgaccggg
    15121 tcacgctgca ctgcaggcat gcaagcttca ccactttgta ccatggtgct ccactagaat
    15181 tcgagctcgg aggaccaagc ccgttattct gacagttctg gtgctcaaca catttatatt
    15241 tatcaaggag cacattgtta ctcactgcta ggagggaatc gaactaggaa tattgatcag
    15301 aggaactacg agagagctga agataactgc cctctagctc tcactgatct gggtcgcata
    15361 gtgagatgca gcccacgtga gttcagcaac ggtctagcgc tgggctttta ggcccgcatg
    15421 atcgggcttt tgtcgggtgg tcgacgtgtt cacgattggg gagagcaacg cagcagttcc
    15481 tcttagttta gtcccacctc gcctgtccag cagagttctg accggtttat aaactcgctt
    15541 gctgcatcag acttgggtgt agcagatgag catgcgtttt agagctagaa atagcaagtt
    15601 aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg ctttttttct
    15661 agacccagct ttcttgtaca aagttggcat tacgctttac gaattcccat ggggaggacc
    15721 aagcccgtta ttctgacagt tctggtgctc aacacattta tatttatcaa ggagcacatt
    15781 gttactcact gctaggaggg aatcgaacta ggaatattga tcagaggaac tacgagagag
    15841 ctgaagataa ctgccctcta gctctcactg atctgggtcg catagtgaga tgcagcccac
    15901 gtgagttcag caacggtcta gcgctgggct tttaggcccg catgatcggg cttttgtcgg
    15961 gtggtcgacg tgttcacgat tggggagagc aacgcagcag ttcctcttag tttagtccca
    16021 cctcgcctgt ccagcagagt tctgaccggt ttataaactc gcttgctgca tcagacttgc
    16081 ttgatgaacc gggaagcgcg ttttagagct agaaatagca agttaaaata aggctagtcc
    16141 gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt ttctagaccc agctttcttg
    16201 tacaaagttg gcattacgct cagagaattc gcatgcggag gaccaagccc gttattctga
    16261 cagttctggt gctcaacaca tttatattta tcaaggagca cattgttact cactgctagg
    16321 agggaatcga actaggaata ttgatcagag gaactacgag agagctgaag ataactgccc
    16381 tctagctctc actgatctgg gtcgcatagt gagatgcagc ccacgtgagt tcagcaacgg
    16441 tctagcgctg ggcttttagg cccgcatgat cgggcttttg tcgggtggtc gacgtgttca
    16501 cgattgggga gagcaacgca gcagttcctc ttagtttagt cccacctcgc ctgtccagca
    16561 gagttctgac cggtttataa actcgcttgc tgcatcagac ttgggacggc accaccgtga
    16621 accgttttag agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa
    16681 agtggcaccg agtcggtgct ttttttctag acccagcttt cttgtacaaa gttggcatta
    16741 cgcttgtgta attcctcgag ggaggaccaa gcccgttatt ctgacagttc tggtgctcaa
    16801 cacatttata tttatcaagg agcacattgt tactcactgc taggagggaa tcgaactagg
    16861 aatattgatc agaggaacta cgagagagct gaagataact gccctctagc tctcactgat
    16921 ctgggtcgca tagtgagatg cagcccacgt gagttcagca acggtctagc gctgggcttt
    16981 taggcccgca tgatcgggct tttgtcgggt ggtcgacgtg ttcacgattg gggagagcaa
    17041 cgcagcagtt cctcttagtt tagtcccacc tcgcctgtcc agcagagttc tgaccggttt
    17101 ataaactcgc ttgctgcatc agacttgcac ggtgaatttg ggcctttgtt ttagagctag
    17161 aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg
    17221 tgcttttttt ctagacccag ctttcttgta caaagttggc attacgctga gcgaggatgc
    17281 acatgtgacc gaggga
//
