LOCUS       sequence-305923-       11601 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing base editor A3A-Delta-186-BE3 in yeast cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11601)
  AUTHORS   Tan J, Zhang F, Karcher D, Bock R
  TITLE     Expanding the genome-targeting scope and the site selectivity of 
            high-precision base editors.
  JOURNAL   Nat Commun. 2020 Jan 31;11(1):629. doi: 10.1038/s41467-020-14465-z.
  PUBMED    32005820
REFERENCE   2  (bases 1 to 11601)
  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: "2020-01-31"; volume: "11(1):629. doi"; pages: " 
            10.1038/s41467-020-14465-z"
COMMENT     Sequence Label: pJT169_GalL_A3A-Delta-186-BE3
FEATURES             Location/Qualifiers
     source          1..11601
                     /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(2071..2097,2098..2100,2101..6174)
                     /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             2071..6174
                     /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             6187..6435
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6448..6468
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      6484..6731
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(6500..6518)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(6749..6768)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6750..6768)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6778..6795)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6778..6794)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6787..6809)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6935..7390
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7022..7041)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7232..7253
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7691..8098
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             8099..9193
                     /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(9687..9706)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9806..9828
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(9866..9884)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    9926..10429
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        10466..10570
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10571..10639,10640..11431)
                     /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(10789..10808)
                     /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 aaatggaagc cagcccagca tccgggccca gacacttgat ggatccacac
     1561 atattcactt ccaactttaa caatggcatt ggaaggcata agacctacct gtgctacgaa
     1621 gtggagcgcc tggacaatgg cacctcggtc aagatggacc agcacagggg ctttctacac
     1681 aaccaggcta agaatcttct ctgtggcttt tacggccgcc atgcggagct gcgcttcttg
     1741 gacctggttc cttctttgca gttggacccg gcccagatct acagggtcac ttggttcatc
     1801 tcctggagcc cctgcttctc ctggggctgt gccggggaag tgcgtgcgtt ccttcaggag
     1861 aacacacacg tgagactgcg tatcttcgct gcccgcatct atgattacga ccccctatat
     1921 aaggaggcac tgcaaatgct gcgggatgct ggggcccaag tctccatcat gacctacgat
     1981 gaatttaagc actgctggga cacctttgtg gaccaccagg gatgtccctt ccagccctgg
     2041 gatggactag atgagcacag ccaagccctg atggacaaga agtactccat tgggctcgct
     2101 atcggcacaa acagcgtcgg ttgggccgtc attacggacg agtacaaggt gccgagcaaa
     2161 aaattcaaag ttctgggcaa taccgatcgc cacagcataa agaagaacct cattggcgcc
     2221 ctcctgttcg actccgggga gacggccgaa gccacgcggc tcaaaagaac agcacggcgc
     2281 agatataccc gcagaaagaa tcggatctgc tacctgcagg agatctttag taatgagatg
     2341 gctaaggtgg atgactcttt cttccatagg ctggaggagt cctttttggt ggaggaggat
     2401 aaaaagcacg agcgccaccc aatctttggc aatatcgtgg acgaggtggc gtaccatgaa
     2461 aagtacccaa ccatatatca tctgaggaag aagcttgtag acagtactga taaggctgac
     2521 ttgcggttga tctatctcgc gctggcgcat atgatcaaat ttcggggaca cttcctcatc
     2581 gagggggacc tgaacccaga caacagcgat gtcgacaaac tctttatcca actggttcag
     2641 acttacaatc agcttttcga agagaacccg atcaacgcat ccggagttga cgccaaagca
     2701 atcctgagcg ctaggctgtc caaatcccgg cggctcgaaa acctcatcgc acagctccct
     2761 ggggagaaga agaacggcct gtttggtaat cttatcgccc tgtcactcgg gctgaccccc
     2821 aactttaaat ctaacttcga cctggccgaa gatgccaagc ttcaactgag caaagacacc
     2881 tacgatgatg atctcgacaa tctgctggcc cagatcggcg accagtacgc agaccttttt
     2941 ttggcggcaa agaacctgtc agacgccatt ctgctgagtg atattctgcg agtgaacacg
     3001 gagatcacca aagctccgct gagcgctagt atgatcaagc gctatgatga gcaccaccaa
     3061 gacttgactt tgctgaaggc ccttgtcaga cagcaactgc ctgagaagta caaggaaatt
     3121 ttcttcgatc agtctaaaaa tggctacgcc ggatacattg acggcggagc aagccaggag
     3181 gaattttaca aatttattaa gcccatcttg gaaaaaatgg acggcaccga ggagctgctg
     3241 gtaaagctta acagagaaga tctgttgcgc aaacagcgca ctttcgacaa tggaagcatc
     3301 ccccaccaga ttcacctggg cgaactgcac gctatcctca ggcggcaaga ggatttctac
     3361 ccctttttga aagataacag ggaaaagatt gagaaaatcc tcacatttcg gataccctac
     3421 tatgtaggcc ccctcgcccg gggaaattcc agattcgcgt ggatgactcg caaatcagaa
     3481 gagaccatca ctccctggaa cttcgaggaa gtcgtggata agggggcctc tgcccagtcc
     3541 ttcatcgaaa ggatgactaa ctttgataaa aatctgccta acgaaaaggt gcttcctaaa
     3601 cactctctgc tgtacgagta cttcacagtt tataacgagc tcaccaaggt caaatacgtc
     3661 acagaaggga tgagaaagcc agcattcctg tctggagagc agaagaaagc tatcgtggac
     3721 ctcctcttca agacgaaccg gaaagttacc gtgaaacagc tcaaagaaga ctatttcaaa
     3781 aagattgaat gtttcgactc tgttgaaatc agcggagtgg aggatcgctt caacgcatcc
     3841 ctgggaacgt atcacgatct cctgaaaatc attaaagaca aggacttcct ggacaatgag
     3901 gagaacgagg acattcttga ggacattgtc ctcaccctta cgttgtttga agatagggag
     3961 atgattgaag aacgcttgaa aacttacgct catctcttcg acgacaaagt catgaaacag
     4021 ctcaagaggc gccgatatac aggatggggg cggctgtcaa gaaaactgat caatgggatc
     4081 cgagacaagc agagtggaaa gacaatcctg gattttctta agtccgatgg atttgccaac
     4141 cggaacttca tgcagttgat ccatgatgac tctctcacct ttaaggagga catccagaaa
     4201 gcacaagttt ctggccaggg ggacagtctt cacgagcaca tcgctaatct tgcaggtagc
     4261 ccagctatca aaaagggaat actgcagacc gttaaggtcg tggatgaact cgtcaaagta
     4321 atgggaaggc ataagcccga gaatatcgtt atcgagatgg cccgagagaa ccaaactacc
     4381 cagaagggac agaagaacag tagggaaagg atgaagagga ttgaagaggg tataaaagaa
     4441 ctggggtccc aaatccttaa ggaacaccca gttgaaaaca cccagcttca gaatgagaag
     4501 ctctacctgt actacctgca gaacggcagg gacatgtacg tggatcagga actggacatc
     4561 aatcggctct ccgactacga cgtggatcat atcgtgcccc agtcttttct caaagatgat
     4621 tctattgata ataaagtgtt gacaagatcc gataaaaata gagggaagag tgataacgtc
     4681 ccctcagaag aagttgtcaa gaaaatgaaa aattattggc ggcagctgct gaacgccaaa
     4741 ctgatcacac aacggaagtt cgataatctg actaaggctg aacgaggtgg cctgtctgag
     4801 ttggataaag ccggcttcat caaaaggcag cttgttgaga cacgccagat caccaagcac
     4861 gtggcccaaa ttctcgattc acgcatgaac accaagtacg atgaaaatga caaactgatt
     4921 cgagaggtga aagttattac tctgaagtct aagctggtct cagatttcag aaaggacttt
     4981 cagttttata aggtgagaga gatcaacaat taccaccatg cgcatgatgc ctacctgaat
     5041 gcagtggtag gcactgcact tatcaaaaaa tatcccaagc ttgaatctga atttgtttac
     5101 ggagactata aagtgtacga tgttaggaaa atgatcgcaa agtctgagca ggaaataggc
     5161 aaggccaccg ctaagtactt cttttacagc aatattatga attttttcaa gaccgagatt
     5221 acactggcca atggagagat tcggaagcga ccacttatcg aaacaaacgg agaaacagga
     5281 gaaatcgtgt gggacaaggg tagggatttc gcgacagtcc ggaaggtcct gtccatgccg
     5341 caggtgaaca tcgttaaaaa gaccgaagta cagaccggag gcttctccaa ggaaagtatc
     5401 ctcccgaaaa ggaacagcga caagctgatc gcacgcaaaa aagattggga ccccaagaaa
     5461 tacggcggat tcgattctcc tacagtcgct tacagtgtac tggttgtggc caaagtggag
     5521 aaagggaagt ctaaaaaact caaaagcgtc aaggaactgc tgggcatcac aatcatggag
     5581 cgatcaagct tcgaaaaaaa ccccatcgac tttctcgagg cgaaaggata taaagaggtc
     5641 aaaaaagacc tcatcattaa gcttcccaag tactctctct ttgagcttga aaacggccgg
     5701 aaacgaatgc tcgctagtgc gggcgagctg cagaaaggta acgagctggc actgccctct
     5761 aaatacgtta atttcttgta tctggccagc cactatgaaa agctcaaagg gtctcccgaa
     5821 gataatgagc agaagcagct gttcgtggaa caacacaaac actaccttga tgagatcatc
     5881 gagcaaataa gcgaattctc caaaagagtg atcctcgccg acgctaacct cgataaggtg
     5941 ctttctgctt acaataagca cagggataag cccatcaggg agcaggcaga aaacattatc
     6001 cacttgttta ctctgaccaa cttgggcgcg cctgcagcct tcaagtactt cgacaccacc
     6061 atagacagaa agcggtacac ctctacaaag gaggtcctgg acgccacact gattcatcag
     6121 tcaattacgg ggctctatga aacaagaatc gacctctctc agctcggtgg agactctggt
     6181 ggttcaacta atttatctga tattattgaa aaagaaacag gtaaacaatt agttattcaa
     6241 gaatcaattt taatgttgcc agaagaagtt gaagaagtta ttggtaataa accagaatct
     6301 gatattttag ttcatactgc ttatgatgaa tctactgatg aaaatgttat gttattgact
     6361 tcagatgctc cagaatataa accatgggct ttagttattc aagattctaa tggtgaaaat
     6421 aaaattaaaa tgttgtctgg tggttcaccc aagaagaaga ggaaggtgtg atctcttctc
     6481 gagtcatgta attagttatg tcacgcttac attcacgccc tccccccaca tccgctctaa
     6541 ccgaaaagga aggagttaga caacctgaag tctaggtccc tatttatttt tttatagtta
     6601 tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg tacagacgcg
     6661 tgtacgcatg taacattata ctgaaaacct tgcttgagaa ggttttggga cgctcgaagg
     6721 ctttaatttg cggccggtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact
     6781 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     6841 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     6901 ttcccaacag ttgcgcagcc tgaatggcga atggacgcgc cctgtagcgg cgcattaagc
     6961 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     7021 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     7081 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     7141 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     7201 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     7261 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     7321 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     7381 cttacaattt cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     7441 tatcgaccgg tcgaggagaa cttctagtat atctacatac ctaatattat tgccttatta
     7501 aaaatggaat cccaacaatt acatcaaaat ccacattctc ttcaaaatca attgtcctgt
     7561 acttccttgt tcatgtgtgt tcaaaaacgt tatatttata ggataattat actctatttc
     7621 tcaacaagta attggttgtt tggccgagcg gtctaaggcg cctgattcaa gaaatatctt
     7681 gaccgcagtt aactgtggga atactcaggt atcgtaagat gcaagagttc gaatctctta
     7741 gcaaccatta tttttttcct caacataacg agaacacaca ggggcgctat cgcacagaat
     7801 caaattcgat gactggaaat tttttgttaa tttcagaggt cgcctgacgc atataccttt
     7861 ttcaactgaa aaattgggag aaaaaggaaa ggtgagagcg ccggaaccgg cttttcatat
     7921 agaatagaga agcgttcatg actaaatgct tgcatcacaa tacttgaagt tgacaatatt
     7981 atttaaggac ctattgtttt ttccaatagg tggttagcaa tcgtcttact ttctaacttt
     8041 tcttaccttt tacatttcag caatatatat atatatattt caaggatata ccattctaat
     8101 gtctgcccct aagaagatcg tcgttttgcc aggtgaccac gttggtcaag aaatcacagc
     8161 cgaagccatt aaggttctta aagctatttc tgatgttcgt tccaatgtca agttcgattt
     8221 cgaaaatcat ttaattggtg gtgctgctat cgatgctaca ggtgttccac ttccagatga
     8281 ggcgctggaa gcctccaaga aggctgatgc cgttttgtta ggtgctgtgg gtggtcctaa
     8341 atggggtacc ggtagtgtta gacctgaaca aggtttacta aaaatccgta aagaacttca
     8401 attgtacgcc aacttaagac catgtaactt tgcatccgac tctcttttag acttatctcc
     8461 aatcaagcca caatttgcta aaggtactga cttcgttgtt gtcagagaat tagtgggagg
     8521 tatttacttt ggtaagagaa aggaagacga tggtgatggt gtcgcttggg atagtgaaca
     8581 atacaccgtt ccagaagtgc aaagaatcac aagaatggcc gctttcatgg ccctacaaca
     8641 tgagccacca ttgcctattt ggtccttgga taaagctaat gttttggcct cttcaagatt
     8701 atggagaaaa actgtggagg aaaccatcaa gaacgaattc cctacattga aggttcaaca
     8761 tcaattgatt gattctgccg ccatgatcct agttaagaac ccaacccacc taaatggtat
     8821 tataatcacc agcaacatgt ttggtgatat catctccgat gaagcctccg ttatcccagg
     8881 ttccttgggt ttgttgccat ctgcgtcctt ggcctctttg ccagacaaga acaccgcatt
     8941 tggtttgtac gaaccatgcc acggttctgc tccagatttg ccaaagaata aggtcaaccc
     9001 tatcgccact atcttgtctg ctgcaatgat gttgaaattg tcattgaact tgcctgaaga
     9061 aggtaaggcc attgaagatg cagttaaaaa ggttttggat gcaggtatca gaactggtga
     9121 tttaggtggt tccaacagta ccaccgaagt cggtgatgct gtcgccgaag aagttaagaa
     9181 aatccttgct taaaaagatt ctcttttttt atgatatttg tacataaact ttataaatga
     9241 aattcataat agaaacgaca cgaaattaca aaatggaata tgttcatagg gtagacgaaa
     9301 ctatatacgc aatctacata catttatcaa gaaggagaaa aaggaggatg taaaggaata
     9361 caggtaagca aattgatact aatggctcaa cgtgataagg aaaaagaatt gcactttaac
     9421 attaatattg acaaggagga gggcaccaca caaaaagtta ggtgtaacag aaaatcatga
     9481 aactatgatt cctaatttat atattggagg attttctcta aaaaaaaaaa aatacaacaa
     9541 ataaaaaaca ctcaatgacc tgaccatttg atggagttta agtcaatacc ttcttgaacc
     9601 atttcccata atggtgaaag ttccctcaag aattttactc tgtcagaaac ggccttaacg
     9661 acgtagtcga tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     9721 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     9781 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     9841 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     9901 atgataataa tggtttctta ggacggatcg cttgcctgta acttacacgc gcctcgtatc
     9961 ttttaatgat ggaataattt gggaatttac tctgtgttta tttattttta tgttttgtat
    10021 ttggatttta gaaagtaaat aaagaaggta gaagagttac ggaatgaaga aaaaaaaata
    10081 aacaaaggtt taaaaaattt caacaaaaag cgtactttac atatatattt attagacaag
    10141 aaaagcagat taaatagata tacattcgat taacgataag taaaatgtaa aatcacagga
    10201 ttttcgtgtg tggtcttcta cacagacaag atgaaacaat tcggcattaa tacctgagag
    10261 caggaagagc aagataaaag gtagtatttg ttggcgatcc ccctagagtc ttttacatct
    10321 tcggaaaaca aaaactattt tttctttaat ttcttttttt actttctatt tttaatttat
    10381 atatttatat taaaaaattt aaattataat tatttttata gcacgtgatg aaaaggaccc
    10441 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
    10501 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
    10561 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
    10621 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
    10681 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
    10741 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
    10801 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
    10861 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
    10921 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
    10981 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
    11041 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
    11101 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
    11161 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
    11221 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
    11281 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
    11341 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
    11401 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
    11461 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
    11521 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
    11581 agaaaagatc aaaggatctt c
//
