LOCUS       sequence-305802-       11622 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses nCDA1-Delta-193-BE3 in yeast cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11622)
  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 11622)
  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: pJT85_GalL_nCDA1-Delta-193-BE3
FEATURES             Location/Qualifiers
     source          1..11622
                     /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(2092..2118,2119..2121,2122..6195)
                     /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             2092..6195
                     /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             6208..6456
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6469..6489
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      6505..6752
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(6521..6539)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(6770..6789)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6771..6789)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6799..6816)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6799..6815)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6808..6830)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6956..7411
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7043..7062)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7253..7274
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7712..8119
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             8120..9214
                     /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(9708..9727)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9827..9849
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(9887..9905)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    9947..10450
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        10487..10591
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10592..10660,10661..11452)
                     /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(10810..10829)
                     /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 tatggacaag
     2101 aagtactcca ttgggctcgc tatcggcaca aacagcgtcg gttgggccgt cattacggac
     2161 gagtacaagg tgccgagcaa aaaattcaaa gttctgggca ataccgatcg ccacagcata
     2221 aagaagaacc tcattggcgc cctcctgttc gactccgggg agacggccga agccacgcgg
     2281 ctcaaaagaa cagcacggcg cagatatacc cgcagaaaga atcggatctg ctacctgcag
     2341 gagatcttta gtaatgagat ggctaaggtg gatgactctt tcttccatag gctggaggag
     2401 tcctttttgg tggaggagga taaaaagcac gagcgccacc caatctttgg caatatcgtg
     2461 gacgaggtgg cgtaccatga aaagtaccca accatatatc atctgaggaa gaagcttgta
     2521 gacagtactg ataaggctga cttgcggttg atctatctcg cgctggcgca tatgatcaaa
     2581 tttcggggac acttcctcat cgagggggac ctgaacccag acaacagcga tgtcgacaaa
     2641 ctctttatcc aactggttca gacttacaat cagcttttcg aagagaaccc gatcaacgca
     2701 tccggagttg acgccaaagc aatcctgagc gctaggctgt ccaaatcccg gcggctcgaa
     2761 aacctcatcg cacagctccc tggggagaag aagaacggcc tgtttggtaa tcttatcgcc
     2821 ctgtcactcg ggctgacccc caactttaaa tctaacttcg acctggccga agatgccaag
     2881 cttcaactga gcaaagacac ctacgatgat gatctcgaca atctgctggc ccagatcggc
     2941 gaccagtacg cagacctttt tttggcggca aagaacctgt cagacgccat tctgctgagt
     3001 gatattctgc gagtgaacac ggagatcacc aaagctccgc tgagcgctag tatgatcaag
     3061 cgctatgatg agcaccacca agacttgact ttgctgaagg cccttgtcag acagcaactg
     3121 cctgagaagt acaaggaaat tttcttcgat cagtctaaaa atggctacgc cggatacatt
     3181 gacggcggag caagccagga ggaattttac aaatttatta agcccatctt ggaaaaaatg
     3241 gacggcaccg aggagctgct ggtaaagctt aacagagaag atctgttgcg caaacagcgc
     3301 actttcgaca atggaagcat cccccaccag attcacctgg gcgaactgca cgctatcctc
     3361 aggcggcaag aggatttcta cccctttttg aaagataaca gggaaaagat tgagaaaatc
     3421 ctcacatttc ggatacccta ctatgtaggc cccctcgccc ggggaaattc cagattcgcg
     3481 tggatgactc gcaaatcaga agagaccatc actccctgga acttcgagga agtcgtggat
     3541 aagggggcct ctgcccagtc cttcatcgaa aggatgacta actttgataa aaatctgcct
     3601 aacgaaaagg tgcttcctaa acactctctg ctgtacgagt acttcacagt ttataacgag
     3661 ctcaccaagg tcaaatacgt cacagaaggg atgagaaagc cagcattcct gtctggagag
     3721 cagaagaaag ctatcgtgga cctcctcttc aagacgaacc ggaaagttac cgtgaaacag
     3781 ctcaaagaag actatttcaa aaagattgaa tgtttcgact ctgttgaaat cagcggagtg
     3841 gaggatcgct tcaacgcatc cctgggaacg tatcacgatc tcctgaaaat cattaaagac
     3901 aaggacttcc tggacaatga ggagaacgag gacattcttg aggacattgt cctcaccctt
     3961 acgttgtttg aagataggga gatgattgaa gaacgcttga aaacttacgc tcatctcttc
     4021 gacgacaaag tcatgaaaca gctcaagagg cgccgatata caggatgggg gcggctgtca
     4081 agaaaactga tcaatgggat ccgagacaag cagagtggaa agacaatcct ggattttctt
     4141 aagtccgatg gatttgccaa ccggaacttc atgcagttga tccatgatga ctctctcacc
     4201 tttaaggagg acatccagaa agcacaagtt tctggccagg gggacagtct tcacgagcac
     4261 atcgctaatc ttgcaggtag cccagctatc aaaaagggaa tactgcagac cgttaaggtc
     4321 gtggatgaac tcgtcaaagt aatgggaagg cataagcccg agaatatcgt tatcgagatg
     4381 gcccgagaga accaaactac ccagaaggga cagaagaaca gtagggaaag gatgaagagg
     4441 attgaagagg gtataaaaga actggggtcc caaatcctta aggaacaccc agttgaaaac
     4501 acccagcttc agaatgagaa gctctacctg tactacctgc agaacggcag ggacatgtac
     4561 gtggatcagg aactggacat caatcggctc tccgactacg acgtggatca tatcgtgccc
     4621 cagtcttttc tcaaagatga ttctattgat aataaagtgt tgacaagatc cgataaaaat
     4681 agagggaaga gtgataacgt cccctcagaa gaagttgtca agaaaatgaa aaattattgg
     4741 cggcagctgc tgaacgccaa actgatcaca caacggaagt tcgataatct gactaaggct
     4801 gaacgaggtg gcctgtctga gttggataaa gccggcttca tcaaaaggca gcttgttgag
     4861 acacgccaga tcaccaagca cgtggcccaa attctcgatt cacgcatgaa caccaagtac
     4921 gatgaaaatg acaaactgat tcgagaggtg aaagttatta ctctgaagtc taagctggtc
     4981 tcagatttca gaaaggactt tcagttttat aaggtgagag agatcaacaa ttaccaccat
     5041 gcgcatgatg cctacctgaa tgcagtggta ggcactgcac ttatcaaaaa atatcccaag
     5101 cttgaatctg aatttgttta cggagactat aaagtgtacg atgttaggaa aatgatcgca
     5161 aagtctgagc aggaaatagg caaggccacc gctaagtact tcttttacag caatattatg
     5221 aattttttca agaccgagat tacactggcc aatggagaga ttcggaagcg accacttatc
     5281 gaaacaaacg gagaaacagg agaaatcgtg tgggacaagg gtagggattt cgcgacagtc
     5341 cggaaggtcc tgtccatgcc gcaggtgaac atcgttaaaa agaccgaagt acagaccgga
     5401 ggcttctcca aggaaagtat cctcccgaaa aggaacagcg acaagctgat cgcacgcaaa
     5461 aaagattggg accccaagaa atacggcgga ttcgattctc ctacagtcgc ttacagtgta
     5521 ctggttgtgg ccaaagtgga gaaagggaag tctaaaaaac tcaaaagcgt caaggaactg
     5581 ctgggcatca caatcatgga gcgatcaagc ttcgaaaaaa accccatcga ctttctcgag
     5641 gcgaaaggat ataaagaggt caaaaaagac ctcatcatta agcttcccaa gtactctctc
     5701 tttgagcttg aaaacggccg gaaacgaatg ctcgctagtg cgggcgagct gcagaaaggt
     5761 aacgagctgg cactgccctc taaatacgtt aatttcttgt atctggccag ccactatgaa
     5821 aagctcaaag ggtctcccga agataatgag cagaagcagc tgttcgtgga acaacacaaa
     5881 cactaccttg atgagatcat cgagcaaata agcgaattct ccaaaagagt gatcctcgcc
     5941 gacgctaacc tcgataaggt gctttctgct tacaataagc acagggataa gcccatcagg
     6001 gagcaggcag aaaacattat ccacttgttt actctgacca acttgggcgc gcctgcagcc
     6061 ttcaagtact tcgacaccac catagacaga aagcggtaca cctctacaaa ggaggtcctg
     6121 gacgccacac tgattcatca gtcaattacg gggctctatg aaacaagaat cgacctctct
     6181 cagctcggtg gagactctgg tggttcaact aatttatctg atattattga aaaagaaaca
     6241 ggtaaacaat tagttattca agaatcaatt ttaatgttgc cagaagaagt tgaagaagtt
     6301 attggtaata aaccagaatc tgatatttta gttcatactg cttatgatga atctactgat
     6361 gaaaatgtta tgttattgac ttcagatgct ccagaatata aaccatgggc tttagttatt
     6421 caagattcta atggtgaaaa taaaattaaa atgttgtctg gtggttcacc caagaagaag
     6481 aggaaggtgt gatctcttct cgagtcatgt aattagttat gtcacgctta cattcacgcc
     6541 ctccccccac atccgctcta accgaaaagg aaggagttag acaacctgaa gtctaggtcc
     6601 ctatttattt ttttatagtt atgttagtat taagaacgtt atttatattt caaatttttc
     6661 ttttttttct gtacagacgc gtgtacgcat gtaacattat actgaaaacc ttgcttgaga
     6721 aggttttggg acgctcgaag gctttaattt gcggccggta cccaattcgc cctatagtga
     6781 gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     6841 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     6901 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggacgcg
     6961 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     7021 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     7081 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     7141 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     7201 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     7261 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     7321 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     7381 aattttaaca aaatattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc
     7441 tgtgcggtat ttcacaccgc atatcgaccg gtcgaggaga acttctagta tatctacata
     7501 cctaatatta ttgccttatt aaaaatggaa tcccaacaat tacatcaaaa tccacattct
     7561 cttcaaaatc aattgtcctg tacttccttg ttcatgtgtg ttcaaaaacg ttatatttat
     7621 aggataatta tactctattt ctcaacaagt aattggttgt ttggccgagc ggtctaaggc
     7681 gcctgattca agaaatatct tgaccgcagt taactgtggg aatactcagg tatcgtaaga
     7741 tgcaagagtt cgaatctctt agcaaccatt atttttttcc tcaacataac gagaacacac
     7801 aggggcgcta tcgcacagaa tcaaattcga tgactggaaa ttttttgtta atttcagagg
     7861 tcgcctgacg catatacctt tttcaactga aaaattggga gaaaaaggaa aggtgagagc
     7921 gccggaaccg gcttttcata tagaatagag aagcgttcat gactaaatgc ttgcatcaca
     7981 atacttgaag ttgacaatat tatttaagga cctattgttt tttccaatag gtggttagca
     8041 atcgtcttac tttctaactt ttcttacctt ttacatttca gcaatatata tatatatatt
     8101 tcaaggatat accattctaa tgtctgcccc taagaagatc gtcgttttgc caggtgacca
     8161 cgttggtcaa gaaatcacag ccgaagccat taaggttctt aaagctattt ctgatgttcg
     8221 ttccaatgtc aagttcgatt tcgaaaatca tttaattggt ggtgctgcta tcgatgctac
     8281 aggtgttcca cttccagatg aggcgctgga agcctccaag aaggctgatg ccgttttgtt
     8341 aggtgctgtg ggtggtccta aatggggtac cggtagtgtt agacctgaac aaggtttact
     8401 aaaaatccgt aaagaacttc aattgtacgc caacttaaga ccatgtaact ttgcatccga
     8461 ctctctttta gacttatctc caatcaagcc acaatttgct aaaggtactg acttcgttgt
     8521 tgtcagagaa ttagtgggag gtatttactt tggtaagaga aaggaagacg atggtgatgg
     8581 tgtcgcttgg gatagtgaac aatacaccgt tccagaagtg caaagaatca caagaatggc
     8641 cgctttcatg gccctacaac atgagccacc attgcctatt tggtccttgg ataaagctaa
     8701 tgttttggcc tcttcaagat tatggagaaa aactgtggag gaaaccatca agaacgaatt
     8761 ccctacattg aaggttcaac atcaattgat tgattctgcc gccatgatcc tagttaagaa
     8821 cccaacccac ctaaatggta ttataatcac cagcaacatg tttggtgata tcatctccga
     8881 tgaagcctcc gttatcccag gttccttggg tttgttgcca tctgcgtcct tggcctcttt
     8941 gccagacaag aacaccgcat ttggtttgta cgaaccatgc cacggttctg ctccagattt
     9001 gccaaagaat aaggtcaacc ctatcgccac tatcttgtct gctgcaatga tgttgaaatt
     9061 gtcattgaac ttgcctgaag aaggtaaggc cattgaagat gcagttaaaa aggttttgga
     9121 tgcaggtatc agaactggtg atttaggtgg ttccaacagt accaccgaag tcggtgatgc
     9181 tgtcgccgaa gaagttaaga aaatccttgc ttaaaaagat tctctttttt tatgatattt
     9241 gtacataaac tttataaatg aaattcataa tagaaacgac acgaaattac aaaatggaat
     9301 atgttcatag ggtagacgaa actatatacg caatctacat acatttatca agaaggagaa
     9361 aaaggaggat gtaaaggaat acaggtaagc aaattgatac taatggctca acgtgataag
     9421 gaaaaagaat tgcactttaa cattaatatt gacaaggagg agggcaccac acaaaaagtt
     9481 aggtgtaaca gaaaatcatg aaactatgat tcctaattta tatattggag gattttctct
     9541 aaaaaaaaaa aaatacaaca aataaaaaac actcaatgac ctgaccattt gatggagttt
     9601 aagtcaatac cttcttgaac catttcccat aatggtgaaa gttccctcaa gaattttact
     9661 ctgtcagaaa cggccttaac gacgtagtcg atatggtgca ctctcagtac aatctgctct
     9721 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     9781 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     9841 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     9901 ctatttttat aggttaatgt catgataata atggtttctt aggacggatc gcttgcctgt
     9961 aacttacacg cgcctcgtat cttttaatga tggaataatt tgggaattta ctctgtgttt
    10021 atttattttt atgttttgta tttggatttt agaaagtaaa taaagaaggt agaagagtta
    10081 cggaatgaag aaaaaaaaat aaacaaaggt ttaaaaaatt tcaacaaaaa gcgtacttta
    10141 catatatatt tattagacaa gaaaagcaga ttaaatagat atacattcga ttaacgataa
    10201 gtaaaatgta aaatcacagg attttcgtgt gtggtcttct acacagacaa gatgaaacaa
    10261 ttcggcatta atacctgaga gcaggaagag caagataaaa ggtagtattt gttggcgatc
    10321 cccctagagt cttttacatc ttcggaaaac aaaaactatt ttttctttaa tttctttttt
    10381 tactttctat ttttaattta tatatttata ttaaaaaatt taaattataa ttatttttat
    10441 agcacgtgat gaaaaggacc caggtggcac ttttcgggga aatgtgcgcg gaacccctat
    10501 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
    10561 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
    10621 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
    10681 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
    10741 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
    10801 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
    10861 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
    10921 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
    10981 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
    11041 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
    11101 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
    11161 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
    11221 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
    11281 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
    11341 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
    11401 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
    11461 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
    11521 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
    11581 ccactgagcg tcagaccccg tagaaaagat caaaggatct tc
//
