LOCUS       sequence-311692-       12061 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses cCDA1-BE3 in yeast cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12061)
  AUTHORS   Tan J, Zhang F, Karcher D, Bock R
  TITLE     Engineering of high-precision base editors for site-specific single 
            nucleotide replacement.
  JOURNAL   Nat Commun. 2019 Jan 25;10(1):439. doi: 10.1038/s41467-018-08034-8.
  PUBMED    30683865
REFERENCE   2  (bases 1 to 12061)
  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: "2019-01-25"; volume: "10(1):439. doi"; pages: " 
            10.1038/s41467-018-08034-8"
COMMENT     Sequence Label: pJT46_GalL_cCDA1-BE3
FEATURES             Location/Qualifiers
     source          1..12061
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..442
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    1..118
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     413..436
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             join(452..478,479..481,482..4555)
                     /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             452..4555
                     /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             4604..5227
                     /codon_start=1
                     /product="cytidine deaminase from the sea lamprey 
                     Petromyzon marinus (Rogozin et al., 2007)"
                     /label=PmCDA1
                     /translation="MTDAEYVRIHEKLDIYTFKKQFFNNKKSVSHRCYVLFELKRRGER
                     RACFWGYAVNKPQSGTERGIHAEIFSIRKVEEYLRDNPGQFTINWYSSWSPCADCAEKI
                     LEWYNQELRGNGHTLKIWACKLYYEKNARNQIGLWNLRDNGVGLNVMVSEHYQCCRKIF
                     IQSSHNQLNENRWLEKTLKRAEKRRSELSIMIQVKILHTTKSPAV"
     CDS             5246..5494
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5507..5527
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      5561..5748
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(5616..5640)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     terminator      complement(5785..5830)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(5866..5885)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5867..5885)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5892..5909)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5892..5908)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5901..5923)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6049..6504
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6136..6155)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6346..6367
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6805..7212
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             7213..8307
                     /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(8801..8820)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8920..8942
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8980..8998)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    9040..9543
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        9580..9684
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9685..9753,9754..10545)
                     /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(9903..9922)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10716..11304
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11205..11224
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11458..11475
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11592..11613
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11628..11633,11634..11651,11652..11658)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11666..11682
                     /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     11671..11693
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11690..11706
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11690..11706
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        11727..11745
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 cggattagaa gccgccgagc gggtgacagc cctccgaagg aagactctcc tccgtgcgtc
       61 ctcgtcttca ccggtcgcgt tcctgaaacg cagatgtgcc tcgcgccgca ctgctccgaa
      121 caataaagat tctacaatac tagcttttat ggttatgaag aggaaaaatt ggcagtaacc
      181 tggccccaca aaccttcaaa tgaacgaatc aaattaacaa ccataggatg ataatgcgat
      241 tagtttttta gccttatttc tggggtaatt aatcagcgaa gcgatgattt ttgatctatt
      301 aacagatata taaatgcaaa aactgcataa ccactttaac taatactttc aacattttcg
      361 gtttgtatta cttcttattc aaatgtaata aaagtatcaa caaaaaattg ttaatatacc
      421 tctatacttt aacgtcaagg agaaaaaacc catggacaag aagtactcca ttgggctcgc
      481 tatcggcaca aacagcgtcg gttgggccgt cattacggac gagtacaagg tgccgagcaa
      541 aaaattcaaa gttctgggca ataccgatcg ccacagcata aagaagaacc tcattggcgc
      601 cctcctgttc gactccgggg agacggccga agccacgcgg ctcaaaagaa cagcacggcg
      661 cagatatacc cgcagaaaga atcggatctg ctacctgcag gagatcttta gtaatgagat
      721 ggctaaggtg gatgactctt tcttccatag gctggaggag tcctttttgg tggaggagga
      781 taaaaagcac gagcgccacc caatctttgg caatatcgtg gacgaggtgg cgtaccatga
      841 aaagtaccca accatatatc atctgaggaa gaagcttgta gacagtactg ataaggctga
      901 cttgcggttg atctatctcg cgctggcgca tatgatcaaa tttcggggac acttcctcat
      961 cgagggggac ctgaacccag acaacagcga tgtcgacaaa ctctttatcc aactggttca
     1021 gacttacaat cagcttttcg aagagaaccc gatcaacgca tccggagttg acgccaaagc
     1081 aatcctgagc gctaggctgt ccaaatcccg gcggctcgaa aacctcatcg cacagctccc
     1141 tggggagaag aagaacggcc tgtttggtaa tcttatcgcc ctgtcactcg ggctgacccc
     1201 caactttaaa tctaacttcg acctggccga agatgccaag cttcaactga gcaaagacac
     1261 ctacgatgat gatctcgaca atctgctggc ccagatcggc gaccagtacg cagacctttt
     1321 tttggcggca aagaacctgt cagacgccat tctgctgagt gatattctgc gagtgaacac
     1381 ggagatcacc aaagctccgc tgagcgctag tatgatcaag cgctatgatg agcaccacca
     1441 agacttgact ttgctgaagg cccttgtcag acagcaactg cctgagaagt acaaggaaat
     1501 tttcttcgat cagtctaaaa atggctacgc cggatacatt gacggcggag caagccagga
     1561 ggaattttac aaatttatta agcccatctt ggaaaaaatg gacggcaccg aggagctgct
     1621 ggtaaagctt aacagagaag atctgttgcg caaacagcgc actttcgaca atggaagcat
     1681 cccccaccag attcacctgg gcgaactgca cgctatcctc aggcggcaag aggatttcta
     1741 cccctttttg aaagataaca gggaaaagat tgagaaaatc ctcacatttc ggatacccta
     1801 ctatgtaggc cccctcgccc ggggaaattc cagattcgcg tggatgactc gcaaatcaga
     1861 agagaccatc actccctgga acttcgagga agtcgtggat aagggggcct ctgcccagtc
     1921 cttcatcgaa aggatgacta actttgataa aaatctgcct aacgaaaagg tgcttcctaa
     1981 acactctctg ctgtacgagt acttcacagt ttataacgag ctcaccaagg tcaaatacgt
     2041 cacagaaggg atgagaaagc cagcattcct gtctggagag cagaagaaag ctatcgtgga
     2101 cctcctcttc aagacgaacc ggaaagttac cgtgaaacag ctcaaagaag actatttcaa
     2161 aaagattgaa tgtttcgact ctgttgaaat cagcggagtg gaggatcgct tcaacgcatc
     2221 cctgggaacg tatcacgatc tcctgaaaat cattaaagac aaggacttcc tggacaatga
     2281 ggagaacgag gacattcttg aggacattgt cctcaccctt acgttgtttg aagataggga
     2341 gatgattgaa gaacgcttga aaacttacgc tcatctcttc gacgacaaag tcatgaaaca
     2401 gctcaagagg cgccgatata caggatgggg gcggctgtca agaaaactga tcaatgggat
     2461 ccgagacaag cagagtggaa agacaatcct ggattttctt aagtccgatg gatttgccaa
     2521 ccggaacttc atgcagttga tccatgatga ctctctcacc tttaaggagg acatccagaa
     2581 agcacaagtt tctggccagg gggacagtct tcacgagcac atcgctaatc ttgcaggtag
     2641 cccagctatc aaaaagggaa tactgcagac cgttaaggtc gtggatgaac tcgtcaaagt
     2701 aatgggaagg cataagcccg agaatatcgt tatcgagatg gcccgagaga accaaactac
     2761 ccagaaggga cagaagaaca gtagggaaag gatgaagagg attgaagagg gtataaaaga
     2821 actggggtcc caaatcctta aggaacaccc agttgaaaac acccagcttc agaatgagaa
     2881 gctctacctg tactacctgc agaacggcag ggacatgtac gtggatcagg aactggacat
     2941 caatcggctc tccgactacg acgtggatca tatcgtgccc cagtcttttc tcaaagatga
     3001 ttctattgat aataaagtgt tgacaagatc cgataaaaat agagggaaga gtgataacgt
     3061 cccctcagaa gaagttgtca agaaaatgaa aaattattgg cggcagctgc tgaacgccaa
     3121 actgatcaca caacggaagt tcgataatct gactaaggct gaacgaggtg gcctgtctga
     3181 gttggataaa gccggcttca tcaaaaggca gcttgttgag acacgccaga tcaccaagca
     3241 cgtggcccaa attctcgatt cacgcatgaa caccaagtac gatgaaaatg acaaactgat
     3301 tcgagaggtg aaagttatta ctctgaagtc taagctggtc tcagatttca gaaaggactt
     3361 tcagttttat aaggtgagag agatcaacaa ttaccaccat gcgcatgatg cctacctgaa
     3421 tgcagtggta ggcactgcac ttatcaaaaa atatcccaag cttgaatctg aatttgttta
     3481 cggagactat aaagtgtacg atgttaggaa aatgatcgca aagtctgagc aggaaatagg
     3541 caaggccacc gctaagtact tcttttacag caatattatg aattttttca agaccgagat
     3601 tacactggcc aatggagaga ttcggaagcg accacttatc gaaacaaacg gagaaacagg
     3661 agaaatcgtg tgggacaagg gtagggattt cgcgacagtc cggaaggtcc tgtccatgcc
     3721 gcaggtgaac atcgttaaaa agaccgaagt acagaccgga ggcttctcca aggaaagtat
     3781 cctcccgaaa aggaacagcg acaagctgat cgcacgcaaa aaagattggg accccaagaa
     3841 atacggcgga ttcgattctc ctacagtcgc ttacagtgta ctggttgtgg ccaaagtgga
     3901 gaaagggaag tctaaaaaac tcaaaagcgt caaggaactg ctgggcatca caattatgga
     3961 gcgatcaagc ttcgaaaaaa accccatcga ctttctcgag gcgaaaggat ataaagaggt
     4021 caaaaaagac ctcatcatta agcttcccaa gtactctctc tttgagcttg aaaacggccg
     4081 gaaacgaatg ctcgctagtg cgggcgagct gcagaaaggt aacgagctgg cactgccctc
     4141 taaatacgtt aatttcttgt atctggccag ccactatgaa aagctcaaag ggtctcccga
     4201 agataatgag cagaagcagc tgttcgtgga acaacacaaa cactaccttg atgagatcat
     4261 cgagcaaata agcgaattct ccaaaagagt gatcctcgcc gacgctaacc tcgataaggt
     4321 gctttctgct tacaataagc acagggataa gcccatcagg gagcaggcag aaaacattat
     4381 ccacttgttt actctgacca acttgggcgc gcctgcagcc ttcaagtact tcgacaccac
     4441 catagacaga aagcggtaca cctctacaaa ggaggtcctg gacgccacac tgattcatca
     4501 gtcaattacg gggctctatg aaacaagaat cgacctctct cagctcggtg gagactctgg
     4561 ttctgaaact ccaggtactt ctgaatctgc tactccagaa tctatgaccg acgctgagta
     4621 cgtgagaatc catgagaagt tggacatcta cacgtttaag aaacagtttt tcaacaacaa
     4681 aaaatccgtg tcgcatagat gctacgttct ctttgaatta aaacgacggg gtgaacgtag
     4741 agcgtgtttt tggggctatg ctgtgaataa accacagagc gggacagaac gtggcattca
     4801 cgccgaaatc tttagcatta gaaaagtcga agaatacctg cgcgacaacc ccggacaatt
     4861 cacgataaat tggtactcat cctggagtcc ttgtgcagat tgcgctgaaa agatcttaga
     4921 atggtataac caggagctgc gggggaacgg ccacactttg aaaatctggg cttgcaaact
     4981 ctattacgag aaaaatgcga ggaatcaaat tgggctgtgg aacctcagag ataacggggt
     5041 tgggttgaat gtaatggtaa gtgaacacta ccaatgttgc aggaaaatat tcatccaatc
     5101 gtcgcacaat caattgaatg agaatagatg gcttgagaag actttgaagc gagctgaaaa
     5161 acgacggagc gagttgtcca ttatgattca ggtaaaaata ctccacacca ctaagagtcc
     5221 tgctgtttct agatctggtg gttcaactaa tttatctgat attattgaaa aagaaacagg
     5281 taaacaatta gttattcaag aatcaatttt aatgttgcca gaagaagttg aagaagttat
     5341 tggtaataaa ccagaatctg atattttagt tcatactgct tatgatgaat ctactgatga
     5401 aaatgttatg ttattgactt cagatgctcc agaatataaa ccatgggctt tagttattca
     5461 agattctaat ggtgaaaata aaattaaaat gttgtctggt ggttcaccca agaagaagag
     5521 gaaggtgtga tctagaggct ccggataagg atcctaataa gcgaatttct tatgatttat
     5581 gatttttatt attaaataag ttataaaaaa aataagtgta tacaaatttt aaagtgactc
     5641 ttaggtttta aaacgaaaat tcttattctt gagtaactct ttcctgtagg tcaggttgct
     5701 ttctcaggta tagcatgagg tcgctcttat tgaccacacc tctaccggca tgccgagcaa
     5761 atgcctgcaa atcgctcccg ggcaaaaaac cccctcaaga cccgtttaga ggccccaagg
     5821 ggttatgcta tgcataccaa gcttccgcgg tggagctcca attcgcccta tagtgagtcg
     5881 tattacaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     5941 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     6001 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggac gcgccctgta
     6061 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     6121 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     6181 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     6241 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     6301 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     6361 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     6421 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     6481 acaaaatatt aacgcttaca atttcctgat gcggtatttt ctccttacgc atctgtgcgg
     6541 tatttcacac cgcatatcga ccggtcgagg agaacttcta gtatatctac atacctaata
     6601 ttattgcctt attaaaaatg gaatcccaac aattacatca aaatccacat tctcttcaaa
     6661 atcaattgtc ctgtacttcc ttgttcatgt gtgttcaaaa acgttatatt tataggataa
     6721 ttatactcta tttctcaaca agtaattggt tgtttggccg agcggtctaa ggcgcctgat
     6781 tcaagaaata tcttgaccgc agttaactgt gggaatactc aggtatcgta agatgcaaga
     6841 gttcgaatct cttagcaacc attatttttt tcctcaacat aacgagaaca cacaggggcg
     6901 ctatcgcaca gaatcaaatt cgatgactgg aaattttttg ttaatttcag aggtcgcctg
     6961 acgcatatac ctttttcaac tgaaaaattg ggagaaaaag gaaaggtgag agcgccggaa
     7021 ccggcttttc atatagaata gagaagcgtt catgactaaa tgcttgcatc acaatacttg
     7081 aagttgacaa tattatttaa ggacctattg ttttttccaa taggtggtta gcaatcgtct
     7141 tactttctaa cttttcttac cttttacatt tcagcaatat atatatatat atttcaagga
     7201 tataccattc taatgtctgc ccctaagaag atcgtcgttt tgccaggtga ccacgttggt
     7261 caagaaatca cagccgaagc cattaaggtt cttaaagcta tttctgatgt tcgttccaat
     7321 gtcaagttcg atttcgaaaa tcatttaatt ggtggtgctg ctatcgatgc tacaggtgtt
     7381 ccacttccag atgaggcgct ggaagcctcc aagaaggctg atgccgtttt gttaggtgct
     7441 gtgggtggtc ctaaatgggg taccggtagt gttagacctg aacaaggttt actaaaaatc
     7501 cgtaaagaac ttcaattgta cgccaactta agaccatgta actttgcatc cgactctctt
     7561 ttagacttat ctccaatcaa gccacaattt gctaaaggta ctgacttcgt tgttgtcaga
     7621 gaattagtgg gaggtattta ctttggtaag agaaaggaag acgatggtga tggtgtcgct
     7681 tgggatagtg aacaatacac cgttccagaa gtgcaaagaa tcacaagaat ggccgctttc
     7741 atggccctac aacatgagcc accattgcct atttggtcct tggataaagc taatgttttg
     7801 gcctcttcaa gattatggag aaaaactgtg gaggaaacca tcaagaacga gttccctaca
     7861 ttgaaggttc aacatcaatt gattgattct gccgccatga tcctagttaa gaacccaacc
     7921 cacctaaatg gtattataat caccagcaac atgtttggtg atatcatctc cgatgaagcc
     7981 tccgttatcc caggttcctt gggtttgttg ccatctgcgt ccttggcctc tttgccagac
     8041 aagaacaccg catttggttt gtacgaacca tgccacggtt ctgctccaga tttgccaaag
     8101 aataaggtca accctatcgc cactatcttg tctgctgcaa tgatgttgaa attgtcattg
     8161 aacttgcctg aagaaggtaa ggccattgaa gatgcagtta aaaaggtttt ggatgcaggt
     8221 atcagaactg gtgatttagg tggttccaac agtaccaccg aagtcggtga tgctgtcgcc
     8281 gaagaagtta agaaaatcct tgcttaaaaa gattctcttt ttttatgata tttgtacata
     8341 aactttataa atgaaattca taatagaaac gacacgaaat tacaaaatgg aatatgttca
     8401 tagggtagac gaaactatat acgcaatcta catacattta tcaagaagga gaaaaaggag
     8461 gatgtaaagg aatacaggta agcaaattga tactaatggc tcaacgtgat aaggaaaaag
     8521 aattgcactt taacattaat attgacaagg aggagggcac cacacaaaaa gttaggtgta
     8581 acagaaaatc atgaaactat gattcctaat ttatatattg gaggattttc tctaaaaaaa
     8641 aaaaaataca acaaataaaa aacactcaat gacctgacca tttgatggag tttaagtcaa
     8701 taccttcttg aaccatttcc cataatggtg aaagttccct caagaatttt actctgtcag
     8761 aaacggcctt aacgacgtag tcgatatggt gcactctcag tacaatctgc tctgatgccg
     8821 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     8881 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     8941 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     9001 tataggttaa tgtcatgata ataatggttt cttaggacgg atcgcttgcc tgtaacttac
     9061 acgcgcctcg tatcttttaa tgatggaata atttgggaat ttactctgtg tttatttatt
     9121 tttatgtttt gtatttggat tttagaaagt aaataaagaa ggtagaagag ttacggaatg
     9181 aagaaaaaaa aataaacaaa ggtttaaaaa atttcaacaa aaagcgtact ttacatatat
     9241 atttattaga caagaaaagc agattaaata gatatacatt cgattaacga taagtaaaat
     9301 gtaaaatcac aggattttcg tgtgtggtct tctacacaga caagatgaaa caattcggca
     9361 ttaatacctg agagcaggaa gagcaagata aaaggtagta tttgttggcg atccccctag
     9421 agtcttttac atcttcggaa aacaaaaact attttttctt taatttcttt ttttactttc
     9481 tatttttaat ttatatattt atattaaaaa atttaaatta taattatttt tatagcacgt
     9541 gatgaaaagg acccaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     9601 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     9661 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     9721 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     9781 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     9841 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     9901 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     9961 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
    10021 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
    10081 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
    10141 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
    10201 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
    10261 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
    10321 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
    10381 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
    10441 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
    10501 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
    10561 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
    10621 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
    10681 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
    10741 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
    10801 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
    10861 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
    10921 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
    10981 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
    11041 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
    11101 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
    11161 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
    11221 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
    11281 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
    11341 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
    11401 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
    11461 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aactgcctct ccccgcgcgt
    11521 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
    11581 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
    11641 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
    11701 tatgaccatg attacgccaa gctcgaaatt aaccctcact aaagggaaca aaagctggta
    11761 ccgggcccga attctctcct tctcttaggt ggcagagcag gtggagggtc gaccatacta
    11821 gtttcaaaaa ttcttacttt ttttttggat ggacgcaaag aagtttaata atcatattac
    11881 atggcattac caccatatac atatccatat acatatccat atacatatcc atatacatat
    11941 ccatatctaa tcttacttat atgttgtgga aatgtaaaga gccccattat cttagcctaa
    12001 aaaaaccttc tctttggaac tttcagtaat acgcttaact gctcattgct atattgaagt
    12061 a
//
