LOCUS       sequence-305818-       11602 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses nCDA1-Delta-188-BE3 in yeast cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11602)
  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 11602)
  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: pJT92_GalL_nCDA1-Delta-188-BE3
FEATURES             Location/Qualifiers
     source          1..11602
                     /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"
     CDS             join(2072..2098,2099..2101,2102..6175)
                     /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             2072..6175
                     /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             6188..6436
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6449..6469
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      6485..6732
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(6501..6519)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(6750..6769)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6751..6769)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6779..6796)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6779..6795)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6788..6810)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6936..7391
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7023..7042)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7233..7254
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7692..8099
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             8100..9194
                     /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(9688..9707)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9807..9829
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(9867..9885)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    9927..10430
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        10467..10571
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10572..10640,10641..11432)
                     /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(10790..10809)
                     /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 gatcccccgg
     1501 gaaaaaaatg accgacgctg agtacgtgag aatccatgag aagttggaca tctacacgtt
     1561 taagaaacag tttttcaaca acaaaaaatc cgtgtcgcat agatgctacg ttctctttga
     1621 attaaaacga cggggtgaac gtagagcgtg tttttggggc tatgctgtga ataaaccaca
     1681 gagcgggaca gaacgtggca ttcacgccga aatctttagc attagaaaag tcgaagaata
     1741 cctgcgcgac aaccccggac aattcacgat aaattggtac tcatcctgga gtccttgtgc
     1801 agattgcgct gaaaagatct tagaatggta taaccaggag ctgcggggga acggccacac
     1861 tttgaaaatc tgggcttgca aactctatta cgagaaaaat gcgaggaatc aaattgggct
     1921 gtggaacctc agagataacg gggttgggtt gaatgtaatg gtaagtgaac actaccaatg
     1981 ttgcaggaaa atattcatcc aatcgtcgca caatcaattg aatgagaata gatggcttga
     2041 gaagactttg aagcgagctg aaaaacgacg gatggacaag aagtactcca ttgggctcgc
     2101 tatcggcaca aacagcgtcg gttgggccgt cattacggac gagtacaagg tgccgagcaa
     2161 aaaattcaaa gttctgggca ataccgatcg ccacagcata aagaagaacc tcattggcgc
     2221 cctcctgttc gactccgggg agacggccga agccacgcgg ctcaaaagaa cagcacggcg
     2281 cagatatacc cgcagaaaga atcggatctg ctacctgcag gagatcttta gtaatgagat
     2341 ggctaaggtg gatgactctt tcttccatag gctggaggag tcctttttgg tggaggagga
     2401 taaaaagcac gagcgccacc caatctttgg caatatcgtg gacgaggtgg cgtaccatga
     2461 aaagtaccca accatatatc atctgaggaa gaagcttgta gacagtactg ataaggctga
     2521 cttgcggttg atctatctcg cgctggcgca tatgatcaaa tttcggggac acttcctcat
     2581 cgagggggac ctgaacccag acaacagcga tgtcgacaaa ctctttatcc aactggttca
     2641 gacttacaat cagcttttcg aagagaaccc gatcaacgca tccggagttg acgccaaagc
     2701 aatcctgagc gctaggctgt ccaaatcccg gcggctcgaa aacctcatcg cacagctccc
     2761 tggggagaag aagaacggcc tgtttggtaa tcttatcgcc ctgtcactcg ggctgacccc
     2821 caactttaaa tctaacttcg acctggccga agatgccaag cttcaactga gcaaagacac
     2881 ctacgatgat gatctcgaca atctgctggc ccagatcggc gaccagtacg cagacctttt
     2941 tttggcggca aagaacctgt cagacgccat tctgctgagt gatattctgc gagtgaacac
     3001 ggagatcacc aaagctccgc tgagcgctag tatgatcaag cgctatgatg agcaccacca
     3061 agacttgact ttgctgaagg cccttgtcag acagcaactg cctgagaagt acaaggaaat
     3121 tttcttcgat cagtctaaaa atggctacgc cggatacatt gacggcggag caagccagga
     3181 ggaattttac aaatttatta agcccatctt ggaaaaaatg gacggcaccg aggagctgct
     3241 ggtaaagctt aacagagaag atctgttgcg caaacagcgc actttcgaca atggaagcat
     3301 cccccaccag attcacctgg gcgaactgca cgctatcctc aggcggcaag aggatttcta
     3361 cccctttttg aaagataaca gggaaaagat tgagaaaatc ctcacatttc ggatacccta
     3421 ctatgtaggc cccctcgccc ggggaaattc cagattcgcg tggatgactc gcaaatcaga
     3481 agagaccatc actccctgga acttcgagga agtcgtggat aagggggcct ctgcccagtc
     3541 cttcatcgaa aggatgacta actttgataa aaatctgcct aacgaaaagg tgcttcctaa
     3601 acactctctg ctgtacgagt acttcacagt ttataacgag ctcaccaagg tcaaatacgt
     3661 cacagaaggg atgagaaagc cagcattcct gtctggagag cagaagaaag ctatcgtgga
     3721 cctcctcttc aagacgaacc ggaaagttac cgtgaaacag ctcaaagaag actatttcaa
     3781 aaagattgaa tgtttcgact ctgttgaaat cagcggagtg gaggatcgct tcaacgcatc
     3841 cctgggaacg tatcacgatc tcctgaaaat cattaaagac aaggacttcc tggacaatga
     3901 ggagaacgag gacattcttg aggacattgt cctcaccctt acgttgtttg aagataggga
     3961 gatgattgaa gaacgcttga aaacttacgc tcatctcttc gacgacaaag tcatgaaaca
     4021 gctcaagagg cgccgatata caggatgggg gcggctgtca agaaaactga tcaatgggat
     4081 ccgagacaag cagagtggaa agacaatcct ggattttctt aagtccgatg gatttgccaa
     4141 ccggaacttc atgcagttga tccatgatga ctctctcacc tttaaggagg acatccagaa
     4201 agcacaagtt tctggccagg gggacagtct tcacgagcac atcgctaatc ttgcaggtag
     4261 cccagctatc aaaaagggaa tactgcagac cgttaaggtc gtggatgaac tcgtcaaagt
     4321 aatgggaagg cataagcccg agaatatcgt tatcgagatg gcccgagaga accaaactac
     4381 ccagaaggga cagaagaaca gtagggaaag gatgaagagg attgaagagg gtataaaaga
     4441 actggggtcc caaatcctta aggaacaccc agttgaaaac acccagcttc agaatgagaa
     4501 gctctacctg tactacctgc agaacggcag ggacatgtac gtggatcagg aactggacat
     4561 caatcggctc tccgactacg acgtggatca tatcgtgccc cagtcttttc tcaaagatga
     4621 ttctattgat aataaagtgt tgacaagatc cgataaaaat agagggaaga gtgataacgt
     4681 cccctcagaa gaagttgtca agaaaatgaa aaattattgg cggcagctgc tgaacgccaa
     4741 actgatcaca caacggaagt tcgataatct gactaaggct gaacgaggtg gcctgtctga
     4801 gttggataaa gccggcttca tcaaaaggca gcttgttgag acacgccaga tcaccaagca
     4861 cgtggcccaa attctcgatt cacgcatgaa caccaagtac gatgaaaatg acaaactgat
     4921 tcgagaggtg aaagttatta ctctgaagtc taagctggtc tcagatttca gaaaggactt
     4981 tcagttttat aaggtgagag agatcaacaa ttaccaccat gcgcatgatg cctacctgaa
     5041 tgcagtggta ggcactgcac ttatcaaaaa atatcccaag cttgaatctg aatttgttta
     5101 cggagactat aaagtgtacg atgttaggaa aatgatcgca aagtctgagc aggaaatagg
     5161 caaggccacc gctaagtact tcttttacag caatattatg aattttttca agaccgagat
     5221 tacactggcc aatggagaga ttcggaagcg accacttatc gaaacaaacg gagaaacagg
     5281 agaaatcgtg tgggacaagg gtagggattt cgcgacagtc cggaaggtcc tgtccatgcc
     5341 gcaggtgaac atcgttaaaa agaccgaagt acagaccgga ggcttctcca aggaaagtat
     5401 cctcccgaaa aggaacagcg acaagctgat cgcacgcaaa aaagattggg accccaagaa
     5461 atacggcgga ttcgattctc ctacagtcgc ttacagtgta ctggttgtgg ccaaagtgga
     5521 gaaagggaag tctaaaaaac tcaaaagcgt caaggaactg ctgggcatca caatcatgga
     5581 gcgatcaagc ttcgaaaaaa accccatcga ctttctcgag gcgaaaggat ataaagaggt
     5641 caaaaaagac ctcatcatta agcttcccaa gtactctctc tttgagcttg aaaacggccg
     5701 gaaacgaatg ctcgctagtg cgggcgagct gcagaaaggt aacgagctgg cactgccctc
     5761 taaatacgtt aatttcttgt atctggccag ccactatgaa aagctcaaag ggtctcccga
     5821 agataatgag cagaagcagc tgttcgtgga acaacacaaa cactaccttg atgagatcat
     5881 cgagcaaata agcgaattct ccaaaagagt gatcctcgcc gacgctaacc tcgataaggt
     5941 gctttctgct tacaataagc acagggataa gcccatcagg gagcaggcag aaaacattat
     6001 ccacttgttt actctgacca acttgggcgc gcctgcagcc ttcaagtact tcgacaccac
     6061 catagacaga aagcggtaca cctctacaaa ggaggtcctg gacgccacac tgattcatca
     6121 gtcaattacg gggctctatg aaacaagaat cgacctctct cagctcggtg gagactctgg
     6181 tggttcaact aatttatctg atattattga aaaagaaaca ggtaaacaat tagttattca
     6241 agaatcaatt ttaatgttgc cagaagaagt tgaagaagtt attggtaata aaccagaatc
     6301 tgatatttta gttcatactg cttatgatga atctactgat gaaaatgtta tgttattgac
     6361 ttcagatgct ccagaatata aaccatgggc tttagttatt caagattcta atggtgaaaa
     6421 taaaattaaa atgttgtctg gtggttcacc caagaagaag aggaaggtgt gatctcttct
     6481 cgagtcatgt aattagttat gtcacgctta cattcacgcc ctccccccac atccgctcta
     6541 accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt ttttatagtt
     6601 atgttagtat taagaacgtt atttatattt caaatttttc ttttttttct gtacagacgc
     6661 gtgtacgcat gtaacattat actgaaaacc ttgcttgaga aggttttggg acgctcgaag
     6721 gctttaattt gcggccggta cccaattcgc cctatagtga gtcgtattac gcgcgctcac
     6781 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     6841 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     6901 cttcccaaca gttgcgcagc ctgaatggcg aatggacgcg ccctgtagcg gcgcattaag
     6961 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     7021 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     7081 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     7141 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     7201 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     7261 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     7321 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     7381 gcttacaatt tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     7441 atatcgaccg gtcgaggaga acttctagta tatctacata cctaatatta ttgccttatt
     7501 aaaaatggaa tcccaacaat tacatcaaaa tccacattct cttcaaaatc aattgtcctg
     7561 tacttccttg ttcatgtgtg ttcaaaaacg ttatatttat aggataatta tactctattt
     7621 ctcaacaagt aattggttgt ttggccgagc ggtctaaggc gcctgattca agaaatatct
     7681 tgaccgcagt taactgtggg aatactcagg tatcgtaaga tgcaagagtt cgaatctctt
     7741 agcaaccatt atttttttcc tcaacataac gagaacacac aggggcgcta tcgcacagaa
     7801 tcaaattcga tgactggaaa ttttttgtta atttcagagg tcgcctgacg catatacctt
     7861 tttcaactga aaaattggga gaaaaaggaa aggtgagagc gccggaaccg gcttttcata
     7921 tagaatagag aagcgttcat gactaaatgc ttgcatcaca atacttgaag ttgacaatat
     7981 tatttaagga cctattgttt tttccaatag gtggttagca atcgtcttac tttctaactt
     8041 ttcttacctt ttacatttca gcaatatata tatatatatt tcaaggatat accattctaa
     8101 tgtctgcccc taagaagatc gtcgttttgc caggtgacca cgttggtcaa gaaatcacag
     8161 ccgaagccat taaggttctt aaagctattt ctgatgttcg ttccaatgtc aagttcgatt
     8221 tcgaaaatca tttaattggt ggtgctgcta tcgatgctac aggtgttcca cttccagatg
     8281 aggcgctgga agcctccaag aaggctgatg ccgttttgtt aggtgctgtg ggtggtccta
     8341 aatggggtac cggtagtgtt agacctgaac aaggtttact aaaaatccgt aaagaacttc
     8401 aattgtacgc caacttaaga ccatgtaact ttgcatccga ctctctttta gacttatctc
     8461 caatcaagcc acaatttgct aaaggtactg acttcgttgt tgtcagagaa ttagtgggag
     8521 gtatttactt tggtaagaga aaggaagacg atggtgatgg tgtcgcttgg gatagtgaac
     8581 aatacaccgt tccagaagtg caaagaatca caagaatggc cgctttcatg gccctacaac
     8641 atgagccacc attgcctatt tggtccttgg ataaagctaa tgttttggcc tcttcaagat
     8701 tatggagaaa aactgtggag gaaaccatca agaacgaatt ccctacattg aaggttcaac
     8761 atcaattgat tgattctgcc gccatgatcc tagttaagaa cccaacccac ctaaatggta
     8821 ttataatcac cagcaacatg tttggtgata tcatctccga tgaagcctcc gttatcccag
     8881 gttccttggg tttgttgcca tctgcgtcct tggcctcttt gccagacaag aacaccgcat
     8941 ttggtttgta cgaaccatgc cacggttctg ctccagattt gccaaagaat aaggtcaacc
     9001 ctatcgccac tatcttgtct gctgcaatga tgttgaaatt gtcattgaac ttgcctgaag
     9061 aaggtaaggc cattgaagat gcagttaaaa aggttttgga tgcaggtatc agaactggtg
     9121 atttaggtgg ttccaacagt accaccgaag tcggtgatgc tgtcgccgaa gaagttaaga
     9181 aaatccttgc ttaaaaagat tctctttttt tatgatattt gtacataaac tttataaatg
     9241 aaattcataa tagaaacgac acgaaattac aaaatggaat atgttcatag ggtagacgaa
     9301 actatatacg caatctacat acatttatca agaaggagaa aaaggaggat gtaaaggaat
     9361 acaggtaagc aaattgatac taatggctca acgtgataag gaaaaagaat tgcactttaa
     9421 cattaatatt gacaaggagg agggcaccac acaaaaagtt aggtgtaaca gaaaatcatg
     9481 aaactatgat tcctaattta tatattggag gattttctct aaaaaaaaaa aaatacaaca
     9541 aataaaaaac actcaatgac ctgaccattt gatggagttt aagtcaatac cttcttgaac
     9601 catttcccat aatggtgaaa gttccctcaa gaattttact ctgtcagaaa cggccttaac
     9661 gacgtagtcg atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     9721 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     9781 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     9841 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     9901 catgataata atggtttctt aggacggatc gcttgcctgt aacttacacg cgcctcgtat
     9961 cttttaatga tggaataatt tgggaattta ctctgtgttt atttattttt atgttttgta
    10021 tttggatttt agaaagtaaa taaagaaggt agaagagtta cggaatgaag aaaaaaaaat
    10081 aaacaaaggt ttaaaaaatt tcaacaaaaa gcgtacttta catatatatt tattagacaa
    10141 gaaaagcaga ttaaatagat atacattcga ttaacgataa gtaaaatgta aaatcacagg
    10201 attttcgtgt gtggtcttct acacagacaa gatgaaacaa ttcggcatta atacctgaga
    10261 gcaggaagag caagataaaa ggtagtattt gttggcgatc cccctagagt cttttacatc
    10321 ttcggaaaac aaaaactatt ttttctttaa tttctttttt tactttctat ttttaattta
    10381 tatatttata ttaaaaaatt taaattataa ttatttttat agcacgtgat gaaaaggacc
    10441 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
    10501 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
    10561 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
    10621 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
    10681 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
    10741 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
    10801 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
    10861 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
    10921 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
    10981 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
    11041 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
    11101 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
    11161 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
    11221 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
    11281 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
    11341 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
    11401 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
    11461 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
    11521 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
    11581 tagaaaagat caaaggatct tc
//
