LOCUS       sequence-249630-       12016 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes a wheat codon optimized Cas9 module in forward direction for
            golden gate cloning of genome editing binary plasmids..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12016)
  TITLE     Genome editing in wheat
REFERENCE   2  (bases 1 to 12016)
  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: pICSL11085_GG_Wheat_live_Cas9_for
FEATURES             Location/Qualifiers
     source          1..12016
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1902..1911
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1914..1934
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      1983..1992
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1989..6092
                     /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             1989..6092
                     /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             6096..6122
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6102..6128
                     /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             6141..6161
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6168..6188
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6204..6260
                     /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(6732..6734,6735..6737,6738..7451)
                     /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_feature    7719..7743
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(7899..9047)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     rep_origin      complement(9383..9971)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9463..9482)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10142..10933,10934..11002))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     10765..10784
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11003..11107)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      11194..11825
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    11917..11941
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 tctgtgaaga caagcaagaa ttcaagcttg gagcaccatg gtacaaagtg gtgaagcttg
       61 catgcctgca gtgcagcgtg acccggtcgt gcccctctct agagataatg agcattgcat
      121 gtctaagtta taaaaaatta ccacatattt tttttgtcac acttgtttga agtgcagttt
      181 atctatcttt atacatatat ttaaacttta ctctacgaat aatataatct atagtactac
      241 aataatatca gtgttttaga gaatcatata aatgaacagt tagacatggt ctaaaggaca
      301 attgagtatt ttgacaacag gactctacag ttttatcttt ttagtgtgca tgtgttctcc
      361 tttttttttg caaatagctt cacctatata atacttcatc cattttatta gtacatccat
      421 ttagggttta gggttaatgg tttttataga ctaatttttt tagtacatct attttattct
      481 attttagcct ctaaattaag aaaactaaaa ctctatttta gtttttttat ttaataattt
      541 agatataaaa tagaataaaa taaagtgact aaaaattaaa caaataccct ttaagaaatt
      601 aaaaaaacta aggaaacatt tttcttgttt cgagtagata atgccagcct gttaaacgcc
      661 gtcgacgagt ctaacggaca ccaaccagcg aaccagcagc gtcgcgtcgg gccaagcgaa
      721 gcagacggca cggcatctct gtcgctgcct ctggacccct ctcgagagtt ccgctccacc
      781 gttggacttg ctccgctgtc ggcatccaga aattgcgtgg cggagcggca gacgtgagcc
      841 ggcacggcag gcggcctcct cctcctctca cggcaccggc agctacgggg gattcctttc
      901 ccaccgctcc ttcgctttcc cttcctcgcc cgccgtaata aatagacacc ccctccacac
      961 cctctttccc caacctcgtg ttgttcggag cgcacacaca cacaaccaga tctcccccaa
     1021 atccacccgt cggcacctcc gcttcaaggt acgccgctcg tcctcccccc ccccccctct
     1081 ctaccttctc tagatcggcg ttccggtcca tggttagggc ccggtagttc tacttctgtt
     1141 catgtttgtg ttagatccgt gtttgtgtta gatccgtgct gctagcgttc gtacacggat
     1201 gcgacctgta cgtcagacac gttctgattg ctaacttgcc agtgtttctc tttggggaat
     1261 cctgggatgg ctctagccgt tccgcagacg ggatcgattt catgattttt tttgtttcgt
     1321 tgcatagggt ttggtttgcc cttttccttt atttcaatat atgccgtgca cttgtttgtc
     1381 gggtcatctt ttcatgcttt tttttgtctt ggttgtgatg atgtggtgtg gttgggcggt
     1441 cgttcattcg ttctagatcg gagtagaata ctgtttcaaa ctacctggtg tatttattaa
     1501 ttttggaact gtatgtgtgt gtcatacatc ttcatagtta cgagtttaag atggatggaa
     1561 atatcgatct aggataggta tacatgttga tgtgggtttt actgatgcat atacatgatg
     1621 gcatatgcag catctattca tatgctctaa ccttgagtac ctatctatta taataaacaa
     1681 gtatgtttta taattatttt gatcttgata tacttggatg atggcatatg cagcagctat
     1741 atgtggattt ttttagccct gccttcatac gctatttatt tgcttggtac tgtttctttt
     1801 gtcgatgctc accctgttgt ttggtgttac ttctgcaggt cgactctaga ggatctggaa
     1861 ttcccgggga gaatctggag ctctcgagat ttctcacgcg tgccaccatg gggccgaaga
     1921 agaagaggaa ggtgggtagg gtgtgtcgga ttagcagcct tcggtacagg ggtcctggca
     1981 ttgccaccat ggacaagaag tactctatcg ggttggatat aggaacgaat agtgtgggtt
     2041 gggctgtcat cactgacgag tacaaggtgc catcgaagaa atttaaagtg ctgggcaaca
     2101 ccgacaggca ctccatcaag aagaacctaa taggtgcgtt gctctttgat tccggggaga
     2161 cagccgaagc gacccgcctc aagcgcacag caagaaggcg ctatacgcgt agaaagaaca
     2221 ggatctgcta cctacaagag atcttttcca acgaaatggc gaaagtggat gactccttct
     2281 tccatcggct cgaggaaagc ttcctcgtag aagaggacaa gaagcacgaa cgccatccga
     2341 tattcgggaa cattgtcgac gaagtcgcat accacgagaa gtatcctacc atctatcacc
     2401 tgaggaagaa gctcgtagat agcactgaca aggctgattt gcgcctcatc tatttggccc
     2461 tggcccatat gattaagttc cgtgggcact tcctcatcga aggcgacctt aatccggaca
     2521 attccgatgt tgacaaactc ttcatccaac tggtccagac atacaatcag ctctttgaag
     2581 agaaccccat caacgcgtca ggagttgatg cgaaggcaat cttgagtgct cggctgtcga
     2641 aaagccggag gctcgaaaat ctgatagccc aactgccagg cgaaaagaag aacggattgt
     2701 ttggcaacct tattgccctt agtctagggc tcacccctaa cttcaagtct aactttgacc
     2761 tggccgagga tgccaagctc caactgtcca aagacacgta cgacgacgat ctcgacaatc
     2821 tgcttgccca aatcggcgac cagtatgcgg accttttcct tgcagcgaag aacttgtcag
     2881 atgcaatcct cttgtccgat attctgcgcg ttaacactga gattaccaag gctcctcttt
     2941 cggcatccat gatcaaacgg tacgacgaac atcaccagga tctcacgctc ctcaaggctc
     3001 tcgtcagaca acagctgccg gagaagtaca aagagatctt cttcgatcaa tcaaagaacg
     3061 ggtatgctgg gtacatcgac ggtggcgcgt cacaggagga gttctacaag tttatcaagc
     3121 ccattctcga gaagatggat ggcactgaag agctcctggt taagctcaat cgcgaggatc
     3181 tactccgcaa gcagcgtacc ttcgacaatg gaagcattcc acaccaaatc catctgggcg
     3241 agcttcacgc catacttcgt aggcaagagg acttttaccc attcctcaaa gacaaccggg
     3301 agaaaatcga gaagatcctc acattcagaa tcccctacta tgtgggccca ttggcaaggg
     3361 gcaactctag gttcgcgtgg atgactagaa aatcggagga aacgattacc ccatggaact
     3421 tcgaggaggt ggtggacaag ggcgcttctg cccaaagctt catcgagagg atgaccaact
     3481 tcgacaagaa tctgccaaac gagaaggtcc tccccaaaca ttccctactg tacgagtact
     3541 tcaccgtcta caacgagctg accaaggtca agtacgttac tgagggtatg aggaaaccgg
     3601 cctttctgtc aggagagcag aagaaggcca tagtggatct ccttttcaag accaatcgca
     3661 aggtcacagt caagcagcta aaggaggact actttaagaa aatcgagtgc tttgactctg
     3721 tggagataag cggcgttgag gatcgcttca acgcctcgct gggaacgtac cacgacttgc
     3781 tcaagatcat taaggataaa gacttcctcg ataacgaaga gaacgaggac atcttggagg
     3841 acatcgttct gacgcttact ctcttcgagg accgcgagat gatcgaagag aggctcaaga
     3901 cgtatgcgca cctctttgac gacaaagtga tgaagcagct gaagcgcagg cgctacacag
     3961 gttggggcag gctgagcagg aagctgatca atggcattcg cgataagcag tcgggcaaga
     4021 ccatcctcga cttcctcaag tccgatgggt tcgcaaacag gaactttatg cagctaatcc
     4081 acgatgactc cttgaccttc aaggaggaca ttcagaaggc tcaggtttca ggccaaggcg
     4141 attcgctcca cgagcatatc gcgaatttgg ccggatctcc agccatcaag aagggtatac
     4201 ttcaaacagt gaaggttgtc gatgagctgg tcaaagtgat gggtagacat aagcccgaga
     4261 atatcgtgat tgagatggct cgggagaacc aaaccacaca gaaggggcag aagaactcca
     4321 gggaaaggat gaagagaatc gaagaaggca tcaaggagct cggatcacaa attctcaagg
     4381 agcatccggt cgaaaacacc caactgcaaa acgagaagct ctacctctac tacctccaaa
     4441 acggacgcga tatgtacgtg gaccaggagc tcgacatcaa tcgcctatct gactacgatg
     4501 tcgatcatat agtaccacaa agcttcctca aagacgactc tatcgacaac aaggtgctga
     4561 ccaggtccga caagaaccgc gggaagtcag acaacgtgcc ttctgaggaa gtcgtcaaga
     4621 agatgaagaa ctactggagg caactcctca atgcaaagct cattacacaa aggaagttcg
     4681 acaaccttac caaggcggag agaggcggcc tcagtgagct agataaggcc ggatttatca
     4741 agcgccaact ggttgagaca cggcaaatca ccaagcacgt tgcacagatc ctggactcac
     4801 gcatgaacac aaaatacgat gagaatgaca agctaatccg ggaagtcaaa gttattactc
     4861 tgaaatctaa actggtgtcc gactttcgga aggacttcca gttctataag gtccgtgaga
     4921 tcaacaacta ccatcacgcc catgatgcct atctcaatgc tgtggttggc actgcactca
     4981 tcaagaaata tcccaagctt gagtcggagt tcgtctacgg tgactacaag gtttatgacg
     5041 taaggaagat gattgcgaag agtgagcagg aaatcggcaa ggccaccgcc aagtacttct
     5101 tttacagcaa cataatgaac ttcttcaaga ctgagatcac cctggccaat ggagagatca
     5161 gaaaacgtcc gctgatagag actaacggag aaactggtga gatagtgtgg gacaagggga
     5221 gagactttgc tactgtccgg aaggtgctaa gcatgccaca agtcaacatc gtcaagaaaa
     5281 cggaagtcca aaccggcggc ttctcgaaag agtcaatcct tccgaagcgc aatagcgata
     5341 agcttatagc ccgtaagaaa gactgggatc caaagaagta cggtggcttt gacagtccta
     5401 cagtggcgta ctccgtgctg gttgttgcca aggtggaaaa ggggaagtcg aagaagctga
     5461 agagcgtaaa ggagttgctc gggattacca ttatggagcg ttccagcttt gagaagaatc
     5521 ccattgattt cttggaggcg aaaggataca aggaagtcaa gaaagatctc atcataaagt
     5581 tgccgaaata cagcttgttc gagctggaga atgggagaaa gaggatgctt gccagcgcag
     5641 gcgagttgca aaaggggaac gagctggcac tcccatccaa gtacgtcaac ttcctgtatc
     5701 tcgcctcaca ctatgagaaa cttaagggtt ccccagagga caatgagcaa aagcagctgt
     5761 ttgtggagca acacaagcac tacctcgatg agatcatcga acagatcagt gagttctcga
     5821 agagggtgat tcttgctgac gctaaccttg acaaggtgtt gtccgcctac aacaagcatc
     5881 gggacaaacc catccgggag caggcagaaa acatcataca cttgttcacg ttgacgaacc
     5941 ttggcgctcc tgcggcgttc aaatattttg acacgacgat tgataggaag cgctatacca
     6001 gcaccaaaga agtactcgat gccaccctaa ttcaccagtc tattacaggc ctgtacgaaa
     6061 cgaggatcga cttgtcgcaa ctaggaggag atgcctatcc gtatgacgtg ccagattacg
     6121 cctcccttgg gtctggttcc cctaagaaga agcggaaagt ggaggatccc aagaagaagc
     6181 gcaaagtcga cggcggatcc ggagccacga acttttccct gctcaagcaa gctggtgatg
     6241 tggaggagaa tcctggccca gctccaggct caacgtccat ggcgaaggca gataagaaac
     6301 cagcggagaa gaaaccggca gagaaaactc cggcagtcga accagcagca gcggcagaga
     6361 agaaaccaaa agccggaaag aaactcccca aggaaccagc cgtcgccgga gacaagaaga
     6421 agaagagatc aaagaagaac gttgagacat acaagatcta catcttcaag gtgttgaagc
     6481 aagttcatcc agacatcgga atctcaagca aagcgatggg aattatgaac agtttcatca
     6541 acgatatttt tgagaagctc gctggtgaat cttcgaagct ggcgaggtac aacaagaagc
     6601 cgacgattac ttccagggag attcagactg cggttagact tgtgttgccc ggtgagttgg
     6661 cgaaacatgc tgtgtctgaa gggactaagg cggttacgaa gtttacgagt tctgctcagg
     6721 gcgcctcttt catggtgtcc aaaggagagg agctctttac aggagttgtg cccattttgg
     6781 tcgagttgga tggcgacgta aatggccaca agttcagcgt gagaggcgaa ggtgaaggtg
     6841 atgccaccaa tggcaagctt accctcaagt tcatctgtac cactgggaaa ctgccggttc
     6901 catggccaac gttggtcacc actctcacat atggtgttca gtgcttttcg aggtatccgg
     6961 accacatgaa acgccatgac ttcttcaagt cagccatgcc tgaggggtat gtccaagaac
     7021 gcaccatcag ctttaaggac gacggaacgt acaagacacg tgcggaggtc aagttcgagg
     7081 gagatacact ggtgaacagg atcgaactca aagggatcga cttcaaagag gacggcaaca
     7141 tactggggca taagctcgag tacaacttca attctcacaa cgtctacatt actgccgaca
     7201 agcagaagaa cggcattaag gcgaacttca aaatccggca taatgtggag gatggtagtg
     7261 tccaactggc tgaccactac cagcagaaca cgccgatagg cgatggccca gtgcttctgc
     7321 ccgacaatca ctacctgtca acccaatccg ttctgtctaa ggaccctaac gaaaagaggg
     7381 atcacatggt gcttctggag tttgtaacgg ctgcaggcat cactcatggg atggatgagc
     7441 tctacaagtg ataactgcgg ccgcagatat cactagtgtt caaacatttg gcaataaagt
     7501 ttcttaagat tgaatcctgt tgccggtctt gcgatgatta tcatataatt tctgttgaat
     7561 tacgttaagc atgtaataat taacatgtaa tgcatgacgt tatttatgag atgggttttt
     7621 atgattagag tcccgcaatt atacatttaa tacgcgatag aaaacaaaat atacgctact
     7681 attgtcttct gcacgaagtg gtttaaacta tcagtgtttg acaggatata ttggcgggta
     7741 aacctaagag aaaagagcgt ttattagaat aatcggatat ttaaaagggc gtgaaaaggt
     7801 ttatccgttc gtccatttgt atgtgcatgc caaccacagg gttccccaga tcaggcgctg
     7861 gctgctgaac ccccagccgg aactgacccc acaaggccct agcgtttgca atgcaccagg
     7921 tcatcattga cccaggcgtg ttccaccagg ccgctgcctc gcaactcttc gcaggcttcg
     7981 ccgacctgct cgcgccactt cttcacgcgg gtggaatccg atccgcacat gaggcggaag
     8041 gtttccagct tgagcgggta cggctcccgg tgcgagctga aatagtcgaa catccgtcgg
     8101 gccgtcggcg acagcttgcg gtacttctcc catatgaatt tcgtgtagtg gtcgccagca
     8161 aacagcacga cgatttcctc gtcgatcagg acctggcaac gggacgtttt cttgccacgg
     8221 tccaggacgc ggaagcggtg cagcagcgac accgattcca ggtgcccaac gcggtcggac
     8281 gtgaagccca tcgccgtcgc ctgtaggcgc gacaggcatt cctcggcctt cgtgtaatac
     8341 cggccattga tcgaccagcc caggtcctgg caaagctcgt agaacgtgaa ggtgatcggc
     8401 tcgccgatag gggtgcgctt cgcgtactcc aacacctgct gccacaccag ttcgtcatcg
     8461 tcggcccgca gctcgacgcc ggtgtaggtg atcttcacgt ccttgttgac gtggaaaatg
     8521 accttgtttt gcagcgcctc gcgcgggatt ttcttgttgc gcgtggtgaa cagggcagag
     8581 cgggccgtgt cgtttggcat cgctcgcatc gtgtccggcc acggcgcaat atcgaacaag
     8641 gaaagctgca tttccttgat ctgctgcttc gtgtgtttca gcaacgcggc ctgcttggcc
     8701 tcgctgacct gttttgccag gtcctcgccg gcggtttttc gcttcttggt cgtcatagtt
     8761 cctcgcgtgt cgatggtcat cgacttcgcc aaacctgccg cctcctgttc aagacgacgc
     8821 gaacgctcca cggcggccga tggcgcgggc agggcagggg gagccagttg cacgctgtcg
     8881 cgctcgatct tggccgtagc ttgctggacc atcgagccga cggactggaa ggtttcgcgg
     8941 ggcgcacgca tgacggtgcg gcttgcgatg gtttcggcat cctcggcgga aaaccccgcg
     9001 tcgatcagtt cttgcctgta tgccttccgg tcaaacgtcc gattcattca ccctccttgc
     9061 gggattgccc cgactcacgc cggggcaatg tgcccttatt cctgatttga cccgcctggt
     9121 gccttggtgt ccagataatc caccttatcg gcaatgaagt cggtcccgta gaccgtctgg
     9181 ccgtccttct cgtacttggt attccgaatc ttgccctgca cgaataccag cgaccccttg
     9241 cccaaatact tgccgtgggc ctcggcctga gagccaaaac acttgatgcg gaagaagtcg
     9301 gtgcgctcct gcttgtcgcc ggcatcgttg cgccacatct aggatctgcc aggaaccgta
     9361 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     9421 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     9481 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     9541 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     9601 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     9661 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     9721 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     9781 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     9841 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     9901 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     9961 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
    10021 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
    10081 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
    10141 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
    10201 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
    10261 ccagtgctgc aatgataccg cgagaaccac gctcaccggc tccagattta tcagcaataa
    10321 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
    10381 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
    10441 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
    10501 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
    10561 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
    10621 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
    10681 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
    10741 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
    10801 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
    10861 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
    10921 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
    10981 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcaag
    11041 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
    11101 ttccgcgcac gaattggcca gcgctgccat ttttggggtg aggccgttcg cggccgaggg
    11161 gcgcagcccc tggggggatg ggaggcccgc gttagcgggc cgggagggtt cgagaagggg
    11221 gggcaccccc cttcggcgtg cgcggtcacg cgcacagggc gcagccctgg ttaaaaacaa
    11281 ggtttataaa tattggttta aaagcaggtt aaaagacagg ttagcggtgg ccgaaaaacg
    11341 ggcggaaacc cttgcaaatg ctggattttc tgcctgtgga cagcccctca aatgtcaata
    11401 ggtgcgcccc tcatctgtca gcactctgcc cctcaagtgt caaggatcgc gcccctcatc
    11461 tgtcagtagt cgcgcccctc aagtgtcaat accgcagggc acttatcccc aggcttgtcc
    11521 acatcatctg tgggaaactc gcgtaaaatc aggcgttttc gccgatttgc gaggctggcc
    11581 agctccacgt cgccggccga aatcgagcct gcccctcatc tgtcaacgcc gcgccgggtg
    11641 agtcggcccc tcaagtgtca acgtccgccc ctcatctgtc agtgagggcc aagttttccg
    11701 cgaggtatcc acaacgccgg cggccgcggt gtctcgcaca cggcttcgac ggcgtttctg
    11761 gcgcgtttgc agggccatag acggccgcca gcccagcggc gagggcaacc agcccggtga
    11821 gcgtcgcaaa ggagatcctg atctgactga tgggctgcct gtatcgagtg gtgattttgt
    11881 gccgagctgc cggtcgggga gctgttggct ggctggtggc aggatatatt gtggtgtaaa
    11941 caaattgacg cttagacaac ttaataacac attgcggacg tttttaatgt actggggtgg
    12001 atgcagtggg ccccac
//
