LOCUS       sequence-305169-       11625 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses nCDA1-Delta-194-BE3 in yeast cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11625)
  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 11625)
  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: pJT84_GalL_nCDA1-Delta-194-BE3
FEATURES             Location/Qualifiers
     source          1..11625
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1010..1030
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1012..1030
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     1439..1462
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     1483..1499
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1483..1499
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             join(2095..2121,2122..2124,2125..6198)
                     /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             2095..6198
                     /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             6211..6459
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6472..6492
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      6508..6755
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(6524..6542)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(6773..6792)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6774..6792)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6802..6819)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6802..6818)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6811..6833)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6959..7414
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7046..7065)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7256..7277
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7715..8122
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             8123..9217
                     /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(9711..9730)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9830..9852
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(9890..9908)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    9950..10453
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        10490..10594
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10595..10663,10664..11455)
                     /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(10813..10832)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     1021 tcactaaagg gaacaaaagc tggagctcgt gacagccctc cgaaggaaga ctctcctccg
     1081 tgcgtcctcg tcttcaccgg tcgcgttcct gaaacgcaga tgtgcctcgc gccgcactgc
     1141 tccgaacaat aaagattcta caatactagc ttttatggtt atgaagagga aaaattggca
     1201 gtaacctggc cccacaaacc ttcaaatgaa cgaatcaaat taacaaccat aggatgataa
     1261 tgcgattagt tttttagcct tatttctggg gtaattaatc agcgaagcga tgatttttga
     1321 tctattaacg gatatataaa tgcaaaaact gcataaccac tttaactaat actttcaaca
     1381 ttttcggttt gtattacttc ttattcaaat gtaataaaag tatcaacaaa aaattgttaa
     1441 tatacctcta tactttaacg tcaaggagaa aaaaccccgg attctagaac tagtggatcc
     1501 cccgggaaaa aaatgaccga cgctgagtac gtgagaatcc atgagaagtt ggacatctac
     1561 acgtttaaga aacagttttt caacaacaaa aaatccgtgt cgcatagatg ctacgttctc
     1621 tttgaattaa aacgacgggg tgaacgtaga gcgtgttttt ggggctatgc tgtgaataaa
     1681 ccacagagcg ggacagaacg tggcattcac gccgaaatct ttagcattag aaaagtcgaa
     1741 gaatacctgc gcgacaaccc cggacaattc acgataaatt ggtactcatc ctggagtcct
     1801 tgtgcagatt gcgctgaaaa gatcttagaa tggtataacc aggagctgcg ggggaacggc
     1861 cacactttga aaatctgggc ttgcaaactc tattacgaga aaaatgcgag gaatcaaatt
     1921 gggctgtgga acctcagaga taacggggtt gggttgaatg taatggtaag tgaacactac
     1981 caatgttgca ggaaaatatt catccaatcg tcgcacaatc aattgaatga gaatagatgg
     2041 cttgagaaga ctttgaagcg agctgaaaaa cgacggagcg agttgtccat tatgatggac
     2101 aagaagtact ccattgggct cgctatcggc acaaacagcg tcggttgggc cgtcattacg
     2161 gacgagtaca aggtgccgag caaaaaattc aaagttctgg gcaataccga tcgccacagc
     2221 ataaagaaga acctcattgg cgccctcctg ttcgactccg gggagacggc cgaagccacg
     2281 cggctcaaaa gaacagcacg gcgcagatat acccgcagaa agaatcggat ctgctacctg
     2341 caggagatct ttagtaatga gatggctaag gtggatgact ctttcttcca taggctggag
     2401 gagtcctttt tggtggagga ggataaaaag cacgagcgcc acccaatctt tggcaatatc
     2461 gtggacgagg tggcgtacca tgaaaagtac ccaaccatat atcatctgag gaagaagctt
     2521 gtagacagta ctgataaggc tgacttgcgg ttgatctatc tcgcgctggc gcatatgatc
     2581 aaatttcggg gacacttcct catcgagggg gacctgaacc cagacaacag cgatgtcgac
     2641 aaactcttta tccaactggt tcagacttac aatcagcttt tcgaagagaa cccgatcaac
     2701 gcatccggag ttgacgccaa agcaatcctg agcgctaggc tgtccaaatc ccggcggctc
     2761 gaaaacctca tcgcacagct ccctggggag aagaagaacg gcctgtttgg taatcttatc
     2821 gccctgtcac tcgggctgac ccccaacttt aaatctaact tcgacctggc cgaagatgcc
     2881 aagcttcaac tgagcaaaga cacctacgat gatgatctcg acaatctgct ggcccagatc
     2941 ggcgaccagt acgcagacct ttttttggcg gcaaagaacc tgtcagacgc cattctgctg
     3001 agtgatattc tgcgagtgaa cacggagatc accaaagctc cgctgagcgc tagtatgatc
     3061 aagcgctatg atgagcacca ccaagacttg actttgctga aggcccttgt cagacagcaa
     3121 ctgcctgaga agtacaagga aattttcttc gatcagtcta aaaatggcta cgccggatac
     3181 attgacggcg gagcaagcca ggaggaattt tacaaattta ttaagcccat cttggaaaaa
     3241 atggacggca ccgaggagct gctggtaaag cttaacagag aagatctgtt gcgcaaacag
     3301 cgcactttcg acaatggaag catcccccac cagattcacc tgggcgaact gcacgctatc
     3361 ctcaggcggc aagaggattt ctaccccttt ttgaaagata acagggaaaa gattgagaaa
     3421 atcctcacat ttcggatacc ctactatgta ggccccctcg cccggggaaa ttccagattc
     3481 gcgtggatga ctcgcaaatc agaagagacc atcactccct ggaacttcga ggaagtcgtg
     3541 gataaggggg cctctgccca gtccttcatc gaaaggatga ctaactttga taaaaatctg
     3601 cctaacgaaa aggtgcttcc taaacactct ctgctgtacg agtacttcac agtttataac
     3661 gagctcacca aggtcaaata cgtcacagaa gggatgagaa agccagcatt cctgtctgga
     3721 gagcagaaga aagctatcgt ggacctcctc ttcaagacga accggaaagt taccgtgaaa
     3781 cagctcaaag aagactattt caaaaagatt gaatgtttcg actctgttga aatcagcgga
     3841 gtggaggatc gcttcaacgc atccctggga acgtatcacg atctcctgaa aatcattaaa
     3901 gacaaggact tcctggacaa tgaggagaac gaggacattc ttgaggacat tgtcctcacc
     3961 cttacgttgt ttgaagatag ggagatgatt gaagaacgct tgaaaactta cgctcatctc
     4021 ttcgacgaca aagtcatgaa acagctcaag aggcgccgat atacaggatg ggggcggctg
     4081 tcaagaaaac tgatcaatgg gatccgagac aagcagagtg gaaagacaat cctggatttt
     4141 cttaagtccg atggatttgc caaccggaac ttcatgcagt tgatccatga tgactctctc
     4201 acctttaagg aggacatcca gaaagcacaa gtttctggcc agggggacag tcttcacgag
     4261 cacatcgcta atcttgcagg tagcccagct atcaaaaagg gaatactgca gaccgttaag
     4321 gtcgtggatg aactcgtcaa agtaatggga aggcataagc ccgagaatat cgttatcgag
     4381 atggcccgag agaaccaaac tacccagaag ggacagaaga acagtaggga aaggatgaag
     4441 aggattgaag agggtataaa agaactgggg tcccaaatcc ttaaggaaca cccagttgaa
     4501 aacacccagc ttcagaatga gaagctctac ctgtactacc tgcagaacgg cagggacatg
     4561 tacgtggatc aggaactgga catcaatcgg ctctccgact acgacgtgga tcatatcgtg
     4621 ccccagtctt ttctcaaaga tgattctatt gataataaag tgttgacaag atccgataaa
     4681 aatagaggga agagtgataa cgtcccctca gaagaagttg tcaagaaaat gaaaaattat
     4741 tggcggcagc tgctgaacgc caaactgatc acacaacgga agttcgataa tctgactaag
     4801 gctgaacgag gtggcctgtc tgagttggat aaagccggct tcatcaaaag gcagcttgtt
     4861 gagacacgcc agatcaccaa gcacgtggcc caaattctcg attcacgcat gaacaccaag
     4921 tacgatgaaa atgacaaact gattcgagag gtgaaagtta ttactctgaa gtctaagctg
     4981 gtctcagatt tcagaaagga ctttcagttt tataaggtga gagagatcaa caattaccac
     5041 catgcgcatg atgcctacct gaatgcagtg gtaggcactg cacttatcaa aaaatatccc
     5101 aagcttgaat ctgaatttgt ttacggagac tataaagtgt acgatgttag gaaaatgatc
     5161 gcaaagtctg agcaggaaat aggcaaggcc accgctaagt acttctttta cagcaatatt
     5221 atgaattttt tcaagaccga gattacactg gccaatggag agattcggaa gcgaccactt
     5281 atcgaaacaa acggagaaac aggagaaatc gtgtgggaca agggtaggga tttcgcgaca
     5341 gtccggaagg tcctgtccat gccgcaggtg aacatcgtta aaaagaccga agtacagacc
     5401 ggaggcttct ccaaggaaag tatcctcccg aaaaggaaca gcgacaagct gatcgcacgc
     5461 aaaaaagatt gggaccccaa gaaatacggc ggattcgatt ctcctacagt cgcttacagt
     5521 gtactggttg tggccaaagt ggagaaaggg aagtctaaaa aactcaaaag cgtcaaggaa
     5581 ctgctgggca tcacaatcat ggagcgatca agcttcgaaa aaaaccccat cgactttctc
     5641 gaggcgaaag gatataaaga ggtcaaaaaa gacctcatca ttaagcttcc caagtactct
     5701 ctctttgagc ttgaaaacgg ccggaaacga atgctcgcta gtgcgggcga gctgcagaaa
     5761 ggtaacgagc tggcactgcc ctctaaatac gttaatttct tgtatctggc cagccactat
     5821 gaaaagctca aagggtctcc cgaagataat gagcagaagc agctgttcgt ggaacaacac
     5881 aaacactacc ttgatgagat catcgagcaa ataagcgaat tctccaaaag agtgatcctc
     5941 gccgacgcta acctcgataa ggtgctttct gcttacaata agcacaggga taagcccatc
     6001 agggagcagg cagaaaacat tatccacttg tttactctga ccaacttggg cgcgcctgca
     6061 gccttcaagt acttcgacac caccatagac agaaagcggt acacctctac aaaggaggtc
     6121 ctggacgcca cactgattca tcagtcaatt acggggctct atgaaacaag aatcgacctc
     6181 tctcagctcg gtggagactc tggtggttca actaatttat ctgatattat tgaaaaagaa
     6241 acaggtaaac aattagttat tcaagaatca attttaatgt tgccagaaga agttgaagaa
     6301 gttattggta ataaaccaga atctgatatt ttagttcata ctgcttatga tgaatctact
     6361 gatgaaaatg ttatgttatt gacttcagat gctccagaat ataaaccatg ggctttagtt
     6421 attcaagatt ctaatggtga aaataaaatt aaaatgttgt ctggtggttc acccaagaag
     6481 aagaggaagg tgtgatctct tctcgagtca tgtaattagt tatgtcacgc ttacattcac
     6541 gccctccccc cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg
     6601 tccctattta tttttttata gttatgttag tattaagaac gttatttata tttcaaattt
     6661 ttcttttttt tctgtacaga cgcgtgtacg catgtaacat tatactgaaa accttgcttg
     6721 agaaggtttt gggacgctcg aaggctttaa tttgcggccg gtacccaatt cgccctatag
     6781 tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     6841 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     6901 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggac
     6961 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     7021 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     7081 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     7141 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     7201 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     7261 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     7321 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     7381 gcgaatttta acaaaatatt aacgcttaca atttcctgat gcggtatttt ctccttacgc
     7441 atctgtgcgg tatttcacac cgcatatcga ccggtcgagg agaacttcta gtatatctac
     7501 atacctaata ttattgcctt attaaaaatg gaatcccaac aattacatca aaatccacat
     7561 tctcttcaaa atcaattgtc ctgtacttcc ttgttcatgt gtgttcaaaa acgttatatt
     7621 tataggataa ttatactcta tttctcaaca agtaattggt tgtttggccg agcggtctaa
     7681 ggcgcctgat tcaagaaata tcttgaccgc agttaactgt gggaatactc aggtatcgta
     7741 agatgcaaga gttcgaatct cttagcaacc attatttttt tcctcaacat aacgagaaca
     7801 cacaggggcg ctatcgcaca gaatcaaatt cgatgactgg aaattttttg ttaatttcag
     7861 aggtcgcctg acgcatatac ctttttcaac tgaaaaattg ggagaaaaag gaaaggtgag
     7921 agcgccggaa ccggcttttc atatagaata gagaagcgtt catgactaaa tgcttgcatc
     7981 acaatacttg aagttgacaa tattatttaa ggacctattg ttttttccaa taggtggtta
     8041 gcaatcgtct tactttctaa cttttcttac cttttacatt tcagcaatat atatatatat
     8101 atttcaagga tataccattc taatgtctgc ccctaagaag atcgtcgttt tgccaggtga
     8161 ccacgttggt caagaaatca cagccgaagc cattaaggtt cttaaagcta tttctgatgt
     8221 tcgttccaat gtcaagttcg atttcgaaaa tcatttaatt ggtggtgctg ctatcgatgc
     8281 tacaggtgtt ccacttccag atgaggcgct ggaagcctcc aagaaggctg atgccgtttt
     8341 gttaggtgct gtgggtggtc ctaaatgggg taccggtagt gttagacctg aacaaggttt
     8401 actaaaaatc cgtaaagaac ttcaattgta cgccaactta agaccatgta actttgcatc
     8461 cgactctctt ttagacttat ctccaatcaa gccacaattt gctaaaggta ctgacttcgt
     8521 tgttgtcaga gaattagtgg gaggtattta ctttggtaag agaaaggaag acgatggtga
     8581 tggtgtcgct tgggatagtg aacaatacac cgttccagaa gtgcaaagaa tcacaagaat
     8641 ggccgctttc atggccctac aacatgagcc accattgcct atttggtcct tggataaagc
     8701 taatgttttg gcctcttcaa gattatggag aaaaactgtg gaggaaacca tcaagaacga
     8761 attccctaca ttgaaggttc aacatcaatt gattgattct gccgccatga tcctagttaa
     8821 gaacccaacc cacctaaatg gtattataat caccagcaac atgtttggtg atatcatctc
     8881 cgatgaagcc tccgttatcc caggttcctt gggtttgttg ccatctgcgt ccttggcctc
     8941 tttgccagac aagaacaccg catttggttt gtacgaacca tgccacggtt ctgctccaga
     9001 tttgccaaag aataaggtca accctatcgc cactatcttg tctgctgcaa tgatgttgaa
     9061 attgtcattg aacttgcctg aagaaggtaa ggccattgaa gatgcagtta aaaaggtttt
     9121 ggatgcaggt atcagaactg gtgatttagg tggttccaac agtaccaccg aagtcggtga
     9181 tgctgtcgcc gaagaagtta agaaaatcct tgcttaaaaa gattctcttt ttttatgata
     9241 tttgtacata aactttataa atgaaattca taatagaaac gacacgaaat tacaaaatgg
     9301 aatatgttca tagggtagac gaaactatat acgcaatcta catacattta tcaagaagga
     9361 gaaaaaggag gatgtaaagg aatacaggta agcaaattga tactaatggc tcaacgtgat
     9421 aaggaaaaag aattgcactt taacattaat attgacaagg aggagggcac cacacaaaaa
     9481 gttaggtgta acagaaaatc atgaaactat gattcctaat ttatatattg gaggattttc
     9541 tctaaaaaaa aaaaaataca acaaataaaa aacactcaat gacctgacca tttgatggag
     9601 tttaagtcaa taccttcttg aaccatttcc cataatggtg aaagttccct caagaatttt
     9661 actctgtcag aaacggcctt aacgacgtag tcgatatggt gcactctcag tacaatctgc
     9721 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     9781 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     9841 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
     9901 cgcctatttt tataggttaa tgtcatgata ataatggttt cttaggacgg atcgcttgcc
     9961 tgtaacttac acgcgcctcg tatcttttaa tgatggaata atttgggaat ttactctgtg
    10021 tttatttatt tttatgtttt gtatttggat tttagaaagt aaataaagaa ggtagaagag
    10081 ttacggaatg aagaaaaaaa aataaacaaa ggtttaaaaa atttcaacaa aaagcgtact
    10141 ttacatatat atttattaga caagaaaagc agattaaata gatatacatt cgattaacga
    10201 taagtaaaat gtaaaatcac aggattttcg tgtgtggtct tctacacaga caagatgaaa
    10261 caattcggca ttaatacctg agagcaggaa gagcaagata aaaggtagta tttgttggcg
    10321 atccccctag agtcttttac atcttcggaa aacaaaaact attttttctt taatttcttt
    10381 ttttactttc tatttttaat ttatatattt atattaaaaa atttaaatta taattatttt
    10441 tatagcacgt gatgaaaagg acccaggtgg cacttttcgg ggaaatgtgc gcggaacccc
    10501 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
    10561 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
    10621 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
    10681 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
    10741 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
    10801 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
    10861 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
    10921 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
    10981 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
    11041 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
    11101 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
    11161 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
    11221 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
    11281 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
    11341 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
    11401 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
    11461 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
    11521 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
    11581 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttc
//
