LOCUS       sequence-306430-       11688 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing base editor A3A-Y130F-BE3 in yeast cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11688)
  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 11688)
  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: pJT174_GalL_A3A-Y130F-BE3
FEATURES             Location/Qualifiers
     source          1..11688
                     /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             1513..2109
                     /codon_start=1
                     /gene="human APOBEC3A"
                     /gene_synonym="A3A"
                     /product="human cytidine deaminase (C to U editing enzyme) 
                     that efficiently edits methylated cytosines (Wang et al., 
                     2018)"
                     /label=APOBEC3A
                     /note="also known as hA3A"
                     /translation="MEASPASGPRHLMDPHIFTSNFNNGIGRHKTYLCYEVERLDNGTS
                     VKMDQHRGFLHNQAKNLLCGFYGRHAELRFLDLVPSLQLDPAQIYRVTWFISWSPCFSW
                     GCAGEVRAFLQENTHVRLRIFAARIFDYDPLYKEALQMLRDAGAQVSIMTYDEFKHCWD
                     TFVDHQGCPFQPWDGLDEHSQALSGRLRAILQNQGN"
     CDS             join(2158..2184,2185..2187,2188..6261)
                     /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             2158..6261
                     /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             6274..6522
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6535..6555
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      6571..6818
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(6587..6605)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(6836..6855)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6837..6855)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6865..6882)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6865..6881)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6874..6896)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7022..7477
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7109..7128)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7319..7340
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7778..8185
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             8186..9280
                     /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(9774..9793)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9893..9915
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(9953..9971)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    10013..10516
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        10553..10657
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10658..10726,10727..11518)
                     /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(10876..10895)
                     /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 ttgattacga ccccctatat
     1921 aaggaggcac tgcaaatgct gcgggatgct ggggcccaag tctccatcat gacctacgat
     1981 gaatttaagc actgctggga cacctttgtg gaccaccagg gatgtccctt ccagccctgg
     2041 gatggactag atgagcacag ccaagccctg agtgggaggc tgcgggccat tctccagaat
     2101 cagggaaact ctggttctga aactccaggt acttctgaat ctgctactcc agaatctatg
     2161 gacaagaagt actccattgg gctcgctatc ggcacaaaca gcgtcggttg ggccgtcatt
     2221 acggacgagt acaaggtgcc gagcaaaaaa ttcaaagttc tgggcaatac cgatcgccac
     2281 agcataaaga agaacctcat tggcgccctc ctgttcgact ccggggagac ggccgaagcc
     2341 acgcggctca aaagaacagc acggcgcaga tatacccgca gaaagaatcg gatctgctac
     2401 ctgcaggaga tctttagtaa tgagatggct aaggtggatg actctttctt ccataggctg
     2461 gaggagtcct ttttggtgga ggaggataaa aagcacgagc gccacccaat ctttggcaat
     2521 atcgtggacg aggtggcgta ccatgaaaag tacccaacca tatatcatct gaggaagaag
     2581 cttgtagaca gtactgataa ggctgacttg cggttgatct atctcgcgct ggcgcatatg
     2641 atcaaatttc ggggacactt cctcatcgag ggggacctga acccagacaa cagcgatgtc
     2701 gacaaactct ttatccaact ggttcagact tacaatcagc ttttcgaaga gaacccgatc
     2761 aacgcatccg gagttgacgc caaagcaatc ctgagcgcta ggctgtccaa atcccggcgg
     2821 ctcgaaaacc tcatcgcaca gctccctggg gagaagaaga acggcctgtt tggtaatctt
     2881 atcgccctgt cactcgggct gacccccaac tttaaatcta acttcgacct ggccgaagat
     2941 gccaagcttc aactgagcaa agacacctac gatgatgatc tcgacaatct gctggcccag
     3001 atcggcgacc agtacgcaga cctttttttg gcggcaaaga acctgtcaga cgccattctg
     3061 ctgagtgata ttctgcgagt gaacacggag atcaccaaag ctccgctgag cgctagtatg
     3121 atcaagcgct atgatgagca ccaccaagac ttgactttgc tgaaggccct tgtcagacag
     3181 caactgcctg agaagtacaa ggaaattttc ttcgatcagt ctaaaaatgg ctacgccgga
     3241 tacattgacg gcggagcaag ccaggaggaa ttttacaaat ttattaagcc catcttggaa
     3301 aaaatggacg gcaccgagga gctgctggta aagcttaaca gagaagatct gttgcgcaaa
     3361 cagcgcactt tcgacaatgg aagcatcccc caccagattc acctgggcga actgcacgct
     3421 atcctcaggc ggcaagagga tttctacccc tttttgaaag ataacaggga aaagattgag
     3481 aaaatcctca catttcggat accctactat gtaggccccc tcgcccgggg aaattccaga
     3541 ttcgcgtgga tgactcgcaa atcagaagag accatcactc cctggaactt cgaggaagtc
     3601 gtggataagg gggcctctgc ccagtccttc atcgaaagga tgactaactt tgataaaaat
     3661 ctgcctaacg aaaaggtgct tcctaaacac tctctgctgt acgagtactt cacagtttat
     3721 aacgagctca ccaaggtcaa atacgtcaca gaagggatga gaaagccagc attcctgtct
     3781 ggagagcaga agaaagctat cgtggacctc ctcttcaaga cgaaccggaa agttaccgtg
     3841 aaacagctca aagaagacta tttcaaaaag attgaatgtt tcgactctgt tgaaatcagc
     3901 ggagtggagg atcgcttcaa cgcatccctg ggaacgtatc acgatctcct gaaaatcatt
     3961 aaagacaagg acttcctgga caatgaggag aacgaggaca ttcttgagga cattgtcctc
     4021 acccttacgt tgtttgaaga tagggagatg attgaagaac gcttgaaaac ttacgctcat
     4081 ctcttcgacg acaaagtcat gaaacagctc aagaggcgcc gatatacagg atgggggcgg
     4141 ctgtcaagaa aactgatcaa tgggatccga gacaagcaga gtggaaagac aatcctggat
     4201 tttcttaagt ccgatggatt tgccaaccgg aacttcatgc agttgatcca tgatgactct
     4261 ctcaccttta aggaggacat ccagaaagca caagtttctg gccaggggga cagtcttcac
     4321 gagcacatcg ctaatcttgc aggtagccca gctatcaaaa agggaatact gcagaccgtt
     4381 aaggtcgtgg atgaactcgt caaagtaatg ggaaggcata agcccgagaa tatcgttatc
     4441 gagatggccc gagagaacca aactacccag aagggacaga agaacagtag ggaaaggatg
     4501 aagaggattg aagagggtat aaaagaactg gggtcccaaa tccttaagga acacccagtt
     4561 gaaaacaccc agcttcagaa tgagaagctc tacctgtact acctgcagaa cggcagggac
     4621 atgtacgtgg atcaggaact ggacatcaat cggctctccg actacgacgt ggatcatatc
     4681 gtgccccagt cttttctcaa agatgattct attgataata aagtgttgac aagatccgat
     4741 aaaaatagag ggaagagtga taacgtcccc tcagaagaag ttgtcaagaa aatgaaaaat
     4801 tattggcggc agctgctgaa cgccaaactg atcacacaac ggaagttcga taatctgact
     4861 aaggctgaac gaggtggcct gtctgagttg gataaagccg gcttcatcaa aaggcagctt
     4921 gttgagacac gccagatcac caagcacgtg gcccaaattc tcgattcacg catgaacacc
     4981 aagtacgatg aaaatgacaa actgattcga gaggtgaaag ttattactct gaagtctaag
     5041 ctggtctcag atttcagaaa ggactttcag ttttataagg tgagagagat caacaattac
     5101 caccatgcgc atgatgccta cctgaatgca gtggtaggca ctgcacttat caaaaaatat
     5161 cccaagcttg aatctgaatt tgtttacgga gactataaag tgtacgatgt taggaaaatg
     5221 atcgcaaagt ctgagcagga aataggcaag gccaccgcta agtacttctt ttacagcaat
     5281 attatgaatt ttttcaagac cgagattaca ctggccaatg gagagattcg gaagcgacca
     5341 cttatcgaaa caaacggaga aacaggagaa atcgtgtggg acaagggtag ggatttcgcg
     5401 acagtccgga aggtcctgtc catgccgcag gtgaacatcg ttaaaaagac cgaagtacag
     5461 accggaggct tctccaagga aagtatcctc ccgaaaagga acagcgacaa gctgatcgca
     5521 cgcaaaaaag attgggaccc caagaaatac ggcggattcg attctcctac agtcgcttac
     5581 agtgtactgg ttgtggccaa agtggagaaa gggaagtcta aaaaactcaa aagcgtcaag
     5641 gaactgctgg gcatcacaat catggagcga tcaagcttcg aaaaaaaccc catcgacttt
     5701 ctcgaggcga aaggatataa agaggtcaaa aaagacctca tcattaagct tcccaagtac
     5761 tctctctttg agcttgaaaa cggccggaaa cgaatgctcg ctagtgcggg cgagctgcag
     5821 aaaggtaacg agctggcact gccctctaaa tacgttaatt tcttgtatct ggccagccac
     5881 tatgaaaagc tcaaagggtc tcccgaagat aatgagcaga agcagctgtt cgtggaacaa
     5941 cacaaacact accttgatga gatcatcgag caaataagcg aattctccaa aagagtgatc
     6001 ctcgccgacg ctaacctcga taaggtgctt tctgcttaca ataagcacag ggataagccc
     6061 atcagggagc aggcagaaaa cattatccac ttgtttactc tgaccaactt gggcgcgcct
     6121 gcagccttca agtacttcga caccaccata gacagaaagc ggtacacctc tacaaaggag
     6181 gtcctggacg ccacactgat tcatcagtca attacggggc tctatgaaac aagaatcgac
     6241 ctctctcagc tcggtggaga ctctggtggt tcaactaatt tatctgatat tattgaaaaa
     6301 gaaacaggta aacaattagt tattcaagaa tcaattttaa tgttgccaga agaagttgaa
     6361 gaagttattg gtaataaacc agaatctgat attttagttc atactgctta tgatgaatct
     6421 actgatgaaa atgttatgtt attgacttca gatgctccag aatataaacc atgggcttta
     6481 gttattcaag attctaatgg tgaaaataaa attaaaatgt tgtctggtgg ttcacccaag
     6541 aagaagagga aggtgtgatc tcttctcgag tcatgtaatt agttatgtca cgcttacatt
     6601 cacgccctcc ccccacatcc gctctaaccg aaaaggaagg agttagacaa cctgaagtct
     6661 aggtccctat ttattttttt atagttatgt tagtattaag aacgttattt atatttcaaa
     6721 tttttctttt ttttctgtac agacgcgtgt acgcatgtaa cattatactg aaaaccttgc
     6781 ttgagaaggt tttgggacgc tcgaaggctt taatttgcgg ccggtaccca attcgcccta
     6841 tagtgagtcg tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa
     6901 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     6961 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     7021 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     7081 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     7141 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     7201 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     7261 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     7321 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     7381 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     7441 aacgcgaatt ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta
     7501 cgcatctgtg cggtatttca caccgcatat cgaccggtcg aggagaactt ctagtatatc
     7561 tacataccta atattattgc cttattaaaa atggaatccc aacaattaca tcaaaatcca
     7621 cattctcttc aaaatcaatt gtcctgtact tccttgttca tgtgtgttca aaaacgttat
     7681 atttatagga taattatact ctatttctca acaagtaatt ggttgtttgg ccgagcggtc
     7741 taaggcgcct gattcaagaa atatcttgac cgcagttaac tgtgggaata ctcaggtatc
     7801 gtaagatgca agagttcgaa tctcttagca accattattt ttttcctcaa cataacgaga
     7861 acacacaggg gcgctatcgc acagaatcaa attcgatgac tggaaatttt ttgttaattt
     7921 cagaggtcgc ctgacgcata tacctttttc aactgaaaaa ttgggagaaa aaggaaaggt
     7981 gagagcgccg gaaccggctt ttcatataga atagagaagc gttcatgact aaatgcttgc
     8041 atcacaatac ttgaagttga caatattatt taaggaccta ttgttttttc caataggtgg
     8101 ttagcaatcg tcttactttc taacttttct taccttttac atttcagcaa tatatatata
     8161 tatatttcaa ggatatacca ttctaatgtc tgcccctaag aagatcgtcg ttttgccagg
     8221 tgaccacgtt ggtcaagaaa tcacagccga agccattaag gttcttaaag ctatttctga
     8281 tgttcgttcc aatgtcaagt tcgatttcga aaatcattta attggtggtg ctgctatcga
     8341 tgctacaggt gttccacttc cagatgaggc gctggaagcc tccaagaagg ctgatgccgt
     8401 tttgttaggt gctgtgggtg gtcctaaatg gggtaccggt agtgttagac ctgaacaagg
     8461 tttactaaaa atccgtaaag aacttcaatt gtacgccaac ttaagaccat gtaactttgc
     8521 atccgactct cttttagact tatctccaat caagccacaa tttgctaaag gtactgactt
     8581 cgttgttgtc agagaattag tgggaggtat ttactttggt aagagaaagg aagacgatgg
     8641 tgatggtgtc gcttgggata gtgaacaata caccgttcca gaagtgcaaa gaatcacaag
     8701 aatggccgct ttcatggccc tacaacatga gccaccattg cctatttggt ccttggataa
     8761 agctaatgtt ttggcctctt caagattatg gagaaaaact gtggaggaaa ccatcaagaa
     8821 cgaattccct acattgaagg ttcaacatca attgattgat tctgccgcca tgatcctagt
     8881 taagaaccca acccacctaa atggtattat aatcaccagc aacatgtttg gtgatatcat
     8941 ctccgatgaa gcctccgtta tcccaggttc cttgggtttg ttgccatctg cgtccttggc
     9001 ctctttgcca gacaagaaca ccgcatttgg tttgtacgaa ccatgccacg gttctgctcc
     9061 agatttgcca aagaataagg tcaaccctat cgccactatc ttgtctgctg caatgatgtt
     9121 gaaattgtca ttgaacttgc ctgaagaagg taaggccatt gaagatgcag ttaaaaaggt
     9181 tttggatgca ggtatcagaa ctggtgattt aggtggttcc aacagtacca ccgaagtcgg
     9241 tgatgctgtc gccgaagaag ttaagaaaat ccttgcttaa aaagattctc tttttttatg
     9301 atatttgtac ataaacttta taaatgaaat tcataataga aacgacacga aattacaaaa
     9361 tggaatatgt tcatagggta gacgaaacta tatacgcaat ctacatacat ttatcaagaa
     9421 ggagaaaaag gaggatgtaa aggaatacag gtaagcaaat tgatactaat ggctcaacgt
     9481 gataaggaaa aagaattgca ctttaacatt aatattgaca aggaggaggg caccacacaa
     9541 aaagttaggt gtaacagaaa atcatgaaac tatgattcct aatttatata ttggaggatt
     9601 ttctctaaaa aaaaaaaaat acaacaaata aaaaacactc aatgacctga ccatttgatg
     9661 gagtttaagt caataccttc ttgaaccatt tcccataatg gtgaaagttc cctcaagaat
     9721 tttactctgt cagaaacggc cttaacgacg tagtcgatat ggtgcactct cagtacaatc
     9781 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     9841 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     9901 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     9961 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagga cggatcgctt
    10021 gcctgtaact tacacgcgcc tcgtatcttt taatgatgga ataatttggg aatttactct
    10081 gtgtttattt atttttatgt tttgtatttg gattttagaa agtaaataaa gaaggtagaa
    10141 gagttacgga atgaagaaaa aaaaataaac aaaggtttaa aaaatttcaa caaaaagcgt
    10201 actttacata tatatttatt agacaagaaa agcagattaa atagatatac attcgattaa
    10261 cgataagtaa aatgtaaaat cacaggattt tcgtgtgtgg tcttctacac agacaagatg
    10321 aaacaattcg gcattaatac ctgagagcag gaagagcaag ataaaaggta gtatttgttg
    10381 gcgatccccc tagagtcttt tacatcttcg gaaaacaaaa actatttttt ctttaatttc
    10441 tttttttact ttctattttt aatttatata tttatattaa aaaatttaaa ttataattat
    10501 ttttatagca cgtgatgaaa aggacccagg tggcactttt cggggaaatg tgcgcggaac
    10561 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
    10621 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
    10681 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
    10741 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
    10801 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
    10861 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
    10921 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
    10981 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
    11041 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
    11101 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
    11161 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
    11221 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
    11281 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
    11341 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
    11401 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
    11461 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
    11521 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
    11581 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
    11641 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttc
//
