LOCUS       sequence-375886-       11822 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing Target-AID2S in yeasts.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11822)
  AUTHORS   Li A, Mitsunobu H, Yoshioka S, Suzuki T, Kondo A, Nishida K
  TITLE     Cytosine base editing systems with minimized off-target effect and 
            molecular size.
  JOURNAL   Nat Commun. 2022 Aug 8;13(1):4531. doi: 10.1038/s41467-022-32157-8.
  PUBMED    35941130
REFERENCE   2  (bases 1 to 11822)
  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: "Nat 
            Commun."; date: "2022-08-8"; volume: "13(1):4531. doi"; pages: " 
            10.1038/s41467-022-32157-8"
COMMENT     Sequence Label: Yeast_Target-AID2S
FEATURES             Location/Qualifiers
     source          1..11822
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    70..573
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        610..714
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(715..783,784..1575)
                     /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     complement(933..952)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1746..2334
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2235..2254
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2488..2505
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2622..2643
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2658..2663,2664..2681,2682..2688)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2696..2712
                     /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)."
     primer_bind     2701..2723
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2720..2736
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2720..2736
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2757..2775
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2852..3516
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    3068..3185
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     3480..3503
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             join(3985..4011,4012..4014,4015..8088)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             3985..8088
                     /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="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             8101..8121
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8128..8382
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="MTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHT
                     AYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     terminator      8413..8600
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(8468..8492)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     terminator      complement(8637..8682)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(8718..8737)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8719..8737)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8744..8761)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8744..8760)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8753..8775)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8901..9356
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8988..9007)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9198..9219
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9657..10064
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             10065..11159
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     primer_bind     complement(11653..11672)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     11772..11794
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttaggacgg atcgcttgcc tgtaacttac acgcgcctcg tatcttttaa tgatggaata
      121 atttgggaat ttactctgtg tttatttatt tttatgtttt gtatttggat tttagaaagt
      181 aaataaagaa ggtagaagag ttacggaatg aagaaaaaaa aataaacaaa ggtttaaaaa
      241 atttcaacaa aaagcgtact ttacatatat atttattaga caagaaaagc agattaaata
      301 gatatacatt cgattaacga taagtaaaat gtaaaatcac aggattttcg tgtgtggtct
      361 tctacacaga caagatgaaa caattcggca ttaatacctg agagcaggaa gagcaagata
      421 aaaggtagta tttgttggcg atccccctag agtcttttac atcttcggaa aacaaaaact
      481 attttttctt taatttcttt ttttactttc tatttttaat ttatatattt atattaaaaa
      541 atttaaatta taattatttt tatagcacgt gatgaaaagg acccaggtgg cacttttcgg
      601 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
      661 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
      721 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
      781 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
      841 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
      901 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
      961 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     1021 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     1081 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     1141 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     1201 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     1261 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     1321 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     1381 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     1441 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     1501 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     1561 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     1621 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     1681 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     1741 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     1801 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     1861 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     1921 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     1981 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2041 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2101 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2161 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2221 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2281 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2341 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     2401 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     2461 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     2521 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     2581 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     2641 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     2701 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gctcgaaatt
     2761 aaccctcact aaagggaaca aaagctggta ccgggcccga attctctcct tctcttaggt
     2821 ggcagagcag gtggagggtc gaccatacta gtttcaaaaa ttcttacttt ttttttggat
     2881 ggacgcaaag aagtttaata atcatattac atggcattac caccatatac atatccatat
     2941 acatatccat atctaatctt acttatatgt tgtggaaatg taaagagccc cattatctta
     3001 gcctaaaaaa accttctctt tggaactttc agtaatacgc ttaactgctc attgctatat
     3061 tgaagtacgg attagaagcc gccgagcggg tgacagccct ccgaaggaag actctcctcc
     3121 gtgcgtcctc gtcttcaccg gtcgcgttcc tgaaacgcag atgtgcctcg cgccgcactg
     3181 ctccgaacaa taaagattct acaatactag cttttatggt tatgaagagg aaaaattggc
     3241 agtaacctgg ccccacaaac cttcaaatga acgaatcaaa ttaacaacca taggatgata
     3301 atgcgattag ttttttagcc ttatttctgg ggtaattaat cagcgaagcg atgatttttg
     3361 atctattaac agatatataa atgcaaaaac tgcataacca ctttaactaa tactttcaac
     3421 attttcggtt tgtattactt cttattcaaa tgtaataaaa gtatcaacaa aaaattgtta
     3481 atatacctct atactttaac gtcaaggaga aaaaaccagc gctcttcgat gtcgcataga
     3541 tgctacgttc tctttgaatt aaaacgacgg ggtgaacgta gagcgtgttt ttggggctat
     3601 gctgtgaata aaccacagag cgggacagaa cgtggcattc acgccgaaat ctttagcatt
     3661 agaaaagtcg aagaatacct gcgcgacaac cccggacaat tcacgataaa ttggtactca
     3721 tcctggagtc cttgtgcaga ttgcgctgaa aagatcttag aatggtataa ccaggagctg
     3781 cgggggaacg gccacacttt gaaaatcgaa gcttgcaaac tctattacga gaaaaatgcg
     3841 aggaatcaaa ttgggctgca aaacctcaga gataacgggg ttgggttgaa tgtaggcgga
     3901 ggtggcagtg gcgggggtgg ctcgggtggg ggaggcagtg gcggaggtgg cagtggcggg
     3961 ggtggctcgg gtgggggagg atccatggac aagaagtact ccattgggct cgctatcggc
     4021 acaaacagcg tcggttgggc cgtcattacg gacgagtaca aggtgccgag caaaaaattc
     4081 aaagttctgg gcaataccga tcgccacagc ataaagaaga acctcattgg cgccctcctg
     4141 ttcgactccg gggagacggc cgaagccacg cggctcaaaa gaacagcacg gcgcagatat
     4201 acccgcagaa agaatcggat ctgctacctg caggagatct ttagtaatga gatggctaag
     4261 gtggatgact ctttcttcca taggctggag gagtcctttt tggtggagga ggataaaaag
     4321 cacgagcgcc acccaatctt tggcaatatc gtggacgagg tggcgtacca tgaaaagtac
     4381 ccaaccatat atcatctgag gaagaagctt gtagacagta ctgataaggc tgacttgcgg
     4441 ttgatctatc tcgcgctggc gcatatgatc aaatttcggg gacacttcct catcgagggg
     4501 gacctgaacc cagacaacag cgatgtcgac aaactcttta tccaactggt tcagacttac
     4561 aatcagcttt tcgaagagaa cccgatcaac gcatccggag ttgacgccaa agcaatcctg
     4621 agcgctaggc tgtccaaatc ccggcggctc gaaaacctca tcgcacagct ccctggggag
     4681 aagaagaacg gcctgtttgg taatcttatc gccctgtcac tcgggctgac ccccaacttt
     4741 aaatctaact tcgacctggc cgaagatgcc aagcttcaac tgagcaaaga cacctacgat
     4801 gatgatctcg acaatctgct ggcccagatc ggcgaccagt acgcagacct ttttttggcg
     4861 gcaaagaacc tgtcagacgc cattctgctg agtgatattc tgcgagtgaa cacggagatc
     4921 accaaagctc cgctgagcgc tagtatgatc aagcgctatg atgagcacca ccaagacttg
     4981 actttgctga aggcccttgt cagacagcaa ctgcctgaga agtacaagga aattttcttc
     5041 gatcagtcta aaaatggcta cgccggatac attgacggcg gagcaagcca ggaggaattt
     5101 tacaaattta ttaagcccat cttggaaaaa atggacggca ccgaggagct gctggtaaag
     5161 cttaacagag aagatctgtt gcgcaaacag cgcactttcg acaatggaag catcccccac
     5221 cagattcacc tgggcgaact gcacgctatc ctcaggcggc aagaggattt ctaccccttt
     5281 ttgaaagata acagggaaaa gattgagaaa atcctcacat ttcggatacc ctactatgta
     5341 ggccccctcg cccggggaaa ttccagattc gcgtggatga ctcgcaaatc agaagagacc
     5401 atcactccct ggaacttcga ggaagtcgtg gataaggggg cctctgccca gtccttcatc
     5461 gaaaggatga ctaactttga taaaaatctg cctaacgaaa aggtgcttcc taaacactct
     5521 ctgctgtacg agtacttcac agtttataac gagctcacca aggtcaaata cgtcacagaa
     5581 gggatgagaa agccagcatt cctgtctgga gagcagaaga aagctatcgt ggacctcctc
     5641 ttcaagacga accggaaagt taccgtgaaa cagctcaaag aagactattt caaaaagatt
     5701 gaatgtttcg actctgttga aatcagcgga gtggaggatc gcttcaacgc atccctggga
     5761 acgtatcacg atctcctgaa aatcattaaa gacaaggact tcctggacaa tgaggagaac
     5821 gaggacattc ttgaggacat tgtcctcacc cttacgttgt ttgaagatag ggagatgatt
     5881 gaagaacgct tgaaaactta cgctcatctc ttcgacgaca aagtcatgaa acagctcaag
     5941 aggcgccgat atacaggatg ggggcggctg tcaagaaaac tgatcaatgg gatccgagac
     6001 aagcagagtg gaaagacaat cctggatttt cttaagtccg atggatttgc caaccggaac
     6061 ttcatgcagt tgatccatga tgactctctc acctttaagg aggacatcca gaaagcacaa
     6121 gtttctggcc agggggacag tcttcacgag cacatcgcta atcttgcagg tagcccagct
     6181 atcaaaaagg gaatactgca gaccgttaag gtcgtggatg aactcgtcaa agtaatggga
     6241 aggcataagc ccgagaatat cgttatcgag atggcccgag agaaccaaac tacccagaag
     6301 ggacagaaga acagtaggga aaggatgaag aggattgaag agggtataaa agaactgggg
     6361 tcccaaatcc ttaaggaaca cccagttgaa aacacccagc ttcagaatga gaagctctac
     6421 ctgtactacc tgcagaacgg cagggacatg tacgtggatc aggaactgga catcaatcgg
     6481 ctctccgact acgacgtgga tcatatcgtg ccccagtctt ttctcaaaga tgattctatt
     6541 gataataaag tgttgacaag atccgataaa aatagaggga agagtgataa cgtcccctca
     6601 gaagaagttg tcaagaaaat gaaaaattat tggcggcagc tgctgaacgc caaactgatc
     6661 acacaacgga agttcgataa tctgactaag gctgaacgag gtggcctgtc tgagttggat
     6721 aaagccggct tcatcaaaag gcagcttgtt gagacacgcc agatcaccaa gcacgtggcc
     6781 caaattctcg attcacgcat gaacaccaag tacgatgaaa atgacaaact gattcgagag
     6841 gtgaaagtta ttactctgaa gtctaagctg gtctcagatt tcagaaagga ctttcagttt
     6901 tataaggtga gagagatcaa caattaccac catgcgcatg atgcctacct gaatgcagtg
     6961 gtaggcactg cacttatcaa aaaatatccc aagcttgaat ctgaatttgt ttacggagac
     7021 tataaagtgt acgatgttag gaaaatgatc gcaaagtctg agcaggaaat aggcaaggcc
     7081 accgctaagt acttctttta cagcaatatt atgaattttt tcaagaccga gattacactg
     7141 gccaatggag agattcggaa gcgaccactt atcgaaacaa acggagaaac aggagaaatc
     7201 gtgtgggaca agggtaggga tttcgcgaca gtccggaagg tcctgtccat gccgcaggtg
     7261 aacatcgtta aaaagaccga agtacagacc ggaggcttct ccaaggaaag tatcctcccg
     7321 aaaaggaaca gcgacaagct gatcgcacgc aaaaaagatt gggaccccaa gaaatacggc
     7381 ggattcgatt ctcctacagt cgcttacagt gtactggttg tggccaaagt ggagaaaggg
     7441 aagtctaaaa aactcaaaag cgtcaaggaa ctgctgggca tcacaatcat ggagcgatca
     7501 agcttcgaaa aaaaccccat cgactttctc gaggcgaaag gatataaaga ggtcaaaaaa
     7561 gacctcatca ttaagcttcc caagtactct ctctttgagc ttgaaaacgg ccggaaacga
     7621 atgctcgcta gtgcgggcga gctgcagaaa ggtaacgagc tggcactgcc ctctaaatac
     7681 gttaatttct tgtatctggc cagccactat gaaaagctca aagggtctcc cgaagataat
     7741 gagcagaagc agctgttcgt ggaacaacac aaacactacc ttgatgagat catcgagcaa
     7801 ataagcgaat tctccaaaag agtgatcctc gccgacgcta acctcgataa ggtgctttct
     7861 gcttacaata agcacaggga taagcccatc agggagcagg cagaaaacat tatccacttg
     7921 tttactctga ccaacttggg cgcgcctgca gccttcaagt acttcgacac caccatagac
     7981 agaaagcggt acacctctac aaaggaggtc ctggacgcca cactgattca tcagtcaatt
     8041 acggggctct atgaaacaag aatcgacctc tctcagctcg gtggagacag cagggctgac
     8101 cccaagaaga agaggaaggt gggtaccatg accaaccttt ccgacatcat agagaaggaa
     8161 acaggcaaac agttggtcat ccaagagtcg atactcatgc ttcctgaaga agttgaggag
     8221 gtcattggga ataagccgga aagtgacatt ctcgtacaca ctgcgtatga tgagagcacc
     8281 gatgagaacg tgatgctgct cacgtcagat gccccagagt acaaaccctg ggctctggtg
     8341 attcaggact ctaatggaga gaacaagatc aagatgctat agtctagagg ctccggataa
     8401 ggatcctaat aagcgaattt cttatgattt atgattttta ttattaaata agttataaaa
     8461 aaaataagtg tatacaaatt ttaaagtgac tcttaggttt taaaacgaaa attcttattc
     8521 ttgagtaact ctttcctgta ggtcaggttg ctttctcagg tatagcatga ggtcgctctt
     8581 attgaccaca cctctaccgg catgccgagc aaatgcctgc aaatcgctcc cgggcaaaaa
     8641 accccctcaa gacccgttta gaggccccaa ggggttatgc tatgcatacc aagcttccgc
     8701 ggtggagctc caattcgccc tatagtgagt cgtattacaa ttcactggcc gtcgttttac
     8761 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     8821 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     8881 gcagcctgaa tggcgaatgg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     8941 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     9001 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     9061 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     9121 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     9181 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     9241 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     9301 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttcctg
     9361 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatc gaccggtcga
     9421 ggagaacttc tagtatatct acatacctaa tattattgcc ttattaaaaa tggaatccca
     9481 acaattacat caaaatccac attctcttca aaatcaattg tcctgtactt ccttgttcat
     9541 gtgtgttcaa aaacgttata tttataggat aattatactc tatttctcaa caagtaattg
     9601 gttgtttggc cgagcggtct aaggcgcctg attcaagaaa tatcttgacc gcagttaact
     9661 gtgggaatac tcaggtatcg taagatgcaa gagttcgaat ctcttagcaa ccattatttt
     9721 tttcctcaac ataacgagaa cacacagggg cgctatcgca cagaatcaaa ttcgatgact
     9781 ggaaattttt tgttaatttc agaggtcgcc tgacgcatat acctttttca actgaaaaat
     9841 tgggagaaaa aggaaaggtg agagcgccgg aaccggcttt tcatatagaa tagagaagcg
     9901 ttcatgacta aatgcttgca tcacaatact tgaagttgac aatattattt aaggacctat
     9961 tgttttttcc aataggtggt tagcaatcgt cttactttct aacttttctt accttttaca
    10021 tttcagcaat atatatatat atatttcaag gatataccat tctaatgtct gcccctaaga
    10081 agatcgtcgt tttgccaggt gaccacgttg gtcaagaaat cacagccgaa gccattaagg
    10141 ttcttaaagc tatttctgat gttcgttcca atgtcaagtt cgatttcgaa aatcatttaa
    10201 ttggtggtgc tgctatcgat gctacaggtg ttccacttcc agatgaggcg ctggaagcct
    10261 ccaagaaggc tgatgccgtt ttgttaggtg ctgtgggtgg tcctaaatgg ggtaccggta
    10321 gtgttagacc tgaacaaggt ttactaaaaa tccgtaaaga acttcaattg tacgccaact
    10381 taagaccatg taactttgca tccgactctc ttttagactt atctccaatc aagccacaat
    10441 ttgctaaagg tactgacttc gttgttgtca gagaattagt gggaggtatt tactttggta
    10501 agagaaagga agacgatggt gatggtgtcg cttgggatag tgaacaatac accgttccag
    10561 aagtgcaaag aatcacaaga atggccgctt tcatggccct acaacatgag ccaccattgc
    10621 ctatttggtc cttggataaa gctaatgttt tggcctcttc aagattatgg agaaaaactg
    10681 tggaggaaac catcaagaac gagttcccta cattgaaggt tcaacatcaa ttgattgatt
    10741 ctgccgccat gatcctagtt aagaacccaa cccacctaaa tggtattata atcaccagca
    10801 acatgtttgg tgatatcatc tccgatgaag cctccgttat cccaggttcc ttgggtttgt
    10861 tgccatctgc gtccttggcc tctttgccag acaagaacac cgcatttggt ttgtacgaac
    10921 catgccacgg ttctgctcca gatttgccaa agaataaggt caaccctatc gccactatct
    10981 tgtctgctgc aatgatgttg aaattgtcat tgaacttgcc tgaagaaggt aaggccattg
    11041 aagatgcagt taaaaaggtt ttggatgcag gtatcagaac tggtgattta ggtggttcca
    11101 acagtaccac cgaagtcggt gatgctgtcg ccgaagaagt taagaaaatc cttgcttaaa
    11161 aagattctct ttttttatga tatttgtaca taaactttat aaatgaaatt cataatagaa
    11221 acgacacgaa attacaaaat ggaatatgtt catagggtag acgaaactat atacgcaatc
    11281 tacatacatt tatcaagaag gagaaaaagg aggatgtaaa ggaatacagg taagcaaatt
    11341 gatactaatg gctcaacgtg ataaggaaaa agaattgcac tttaacatta atattgacaa
    11401 ggaggagggc accacacaaa aagttaggtg taacagaaaa tcatgaaact atgattccta
    11461 atttatatat tggaggattt tctctaaaaa aaaaaaaata caacaaataa aaaacactca
    11521 atgacctgac catttgatgg agtttaagtc aataccttct tgaaccattt cccataatgg
    11581 tgaaagttcc ctcaagaatt ttactctgtc agaaacggcc ttaacgacgt agtcgatatg
    11641 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
    11701 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
    11761 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
    11821 ga
//
