LOCUS       sequence-305933-       11577 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing base editor A3A-Delta-178-BE3 in yeast cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11577)
  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 11577)
  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: pJT171_GalL_A3A-Delta-178-BE3
FEATURES             Location/Qualifiers
     source          1..11577
                     /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(2047..2073,2074..2076,2077..6150)
                     /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             2047..6150
                     /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             6163..6411
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6424..6444
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      6460..6707
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(6476..6494)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(6725..6744)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6726..6744)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6754..6771)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6754..6770)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6763..6785)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6911..7366
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6998..7017)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7208..7229
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7667..8074
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             8075..9169
                     /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(9663..9682)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9782..9804
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(9842..9860)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    9902..10405
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        10442..10546
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10547..10615,10616..11407)
                     /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(10765..10784)
                     /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 gatggaatgg acaagaagta ctccattggg ctcgctatcg gcacaaacag cgtcggttgg
     2101 gccgtcatta cggacgagta caaggtgccg agcaaaaaat tcaaagttct gggcaatacc
     2161 gatcgccaca gcataaagaa gaacctcatt ggcgccctcc tgttcgactc cggggagacg
     2221 gccgaagcca cgcggctcaa aagaacagca cggcgcagat atacccgcag aaagaatcgg
     2281 atctgctacc tgcaggagat ctttagtaat gagatggcta aggtggatga ctctttcttc
     2341 cataggctgg aggagtcctt tttggtggag gaggataaaa agcacgagcg ccacccaatc
     2401 tttggcaata tcgtggacga ggtggcgtac catgaaaagt acccaaccat atatcatctg
     2461 aggaagaagc ttgtagacag tactgataag gctgacttgc ggttgatcta tctcgcgctg
     2521 gcgcatatga tcaaatttcg gggacacttc ctcatcgagg gggacctgaa cccagacaac
     2581 agcgatgtcg acaaactctt tatccaactg gttcagactt acaatcagct tttcgaagag
     2641 aacccgatca acgcatccgg agttgacgcc aaagcaatcc tgagcgctag gctgtccaaa
     2701 tcccggcggc tcgaaaacct catcgcacag ctccctgggg agaagaagaa cggcctgttt
     2761 ggtaatctta tcgccctgtc actcgggctg acccccaact ttaaatctaa cttcgacctg
     2821 gccgaagatg ccaagcttca actgagcaaa gacacctacg atgatgatct cgacaatctg
     2881 ctggcccaga tcggcgacca gtacgcagac ctttttttgg cggcaaagaa cctgtcagac
     2941 gccattctgc tgagtgatat tctgcgagtg aacacggaga tcaccaaagc tccgctgagc
     3001 gctagtatga tcaagcgcta tgatgagcac caccaagact tgactttgct gaaggccctt
     3061 gtcagacagc aactgcctga gaagtacaag gaaattttct tcgatcagtc taaaaatggc
     3121 tacgccggat acattgacgg cggagcaagc caggaggaat tttacaaatt tattaagccc
     3181 atcttggaaa aaatggacgg caccgaggag ctgctggtaa agcttaacag agaagatctg
     3241 ttgcgcaaac agcgcacttt cgacaatgga agcatccccc accagattca cctgggcgaa
     3301 ctgcacgcta tcctcaggcg gcaagaggat ttctacccct ttttgaaaga taacagggaa
     3361 aagattgaga aaatcctcac atttcggata ccctactatg taggccccct cgcccgggga
     3421 aattccagat tcgcgtggat gactcgcaaa tcagaagaga ccatcactcc ctggaacttc
     3481 gaggaagtcg tggataaggg ggcctctgcc cagtccttca tcgaaaggat gactaacttt
     3541 gataaaaatc tgcctaacga aaaggtgctt cctaaacact ctctgctgta cgagtacttc
     3601 acagtttata acgagctcac caaggtcaaa tacgtcacag aagggatgag aaagccagca
     3661 ttcctgtctg gagagcagaa gaaagctatc gtggacctcc tcttcaagac gaaccggaaa
     3721 gttaccgtga aacagctcaa agaagactat ttcaaaaaga ttgaatgttt cgactctgtt
     3781 gaaatcagcg gagtggagga tcgcttcaac gcatccctgg gaacgtatca cgatctcctg
     3841 aaaatcatta aagacaagga cttcctggac aatgaggaga acgaggacat tcttgaggac
     3901 attgtcctca cccttacgtt gtttgaagat agggagatga ttgaagaacg cttgaaaact
     3961 tacgctcatc tcttcgacga caaagtcatg aaacagctca agaggcgccg atatacagga
     4021 tgggggcggc tgtcaagaaa actgatcaat gggatccgag acaagcagag tggaaagaca
     4081 atcctggatt ttcttaagtc cgatggattt gccaaccgga acttcatgca gttgatccat
     4141 gatgactctc tcacctttaa ggaggacatc cagaaagcac aagtttctgg ccagggggac
     4201 agtcttcacg agcacatcgc taatcttgca ggtagcccag ctatcaaaaa gggaatactg
     4261 cagaccgtta aggtcgtgga tgaactcgtc aaagtaatgg gaaggcataa gcccgagaat
     4321 atcgttatcg agatggcccg agagaaccaa actacccaga agggacagaa gaacagtagg
     4381 gaaaggatga agaggattga agagggtata aaagaactgg ggtcccaaat ccttaaggaa
     4441 cacccagttg aaaacaccca gcttcagaat gagaagctct acctgtacta cctgcagaac
     4501 ggcagggaca tgtacgtgga tcaggaactg gacatcaatc ggctctccga ctacgacgtg
     4561 gatcatatcg tgccccagtc ttttctcaaa gatgattcta ttgataataa agtgttgaca
     4621 agatccgata aaaatagagg gaagagtgat aacgtcccct cagaagaagt tgtcaagaaa
     4681 atgaaaaatt attggcggca gctgctgaac gccaaactga tcacacaacg gaagttcgat
     4741 aatctgacta aggctgaacg aggtggcctg tctgagttgg ataaagccgg cttcatcaaa
     4801 aggcagcttg ttgagacacg ccagatcacc aagcacgtgg cccaaattct cgattcacgc
     4861 atgaacacca agtacgatga aaatgacaaa ctgattcgag aggtgaaagt tattactctg
     4921 aagtctaagc tggtctcaga tttcagaaag gactttcagt tttataaggt gagagagatc
     4981 aacaattacc accatgcgca tgatgcctac ctgaatgcag tggtaggcac tgcacttatc
     5041 aaaaaatatc ccaagcttga atctgaattt gtttacggag actataaagt gtacgatgtt
     5101 aggaaaatga tcgcaaagtc tgagcaggaa ataggcaagg ccaccgctaa gtacttcttt
     5161 tacagcaata ttatgaattt tttcaagacc gagattacac tggccaatgg agagattcgg
     5221 aagcgaccac ttatcgaaac aaacggagaa acaggagaaa tcgtgtggga caagggtagg
     5281 gatttcgcga cagtccggaa ggtcctgtcc atgccgcagg tgaacatcgt taaaaagacc
     5341 gaagtacaga ccggaggctt ctccaaggaa agtatcctcc cgaaaaggaa cagcgacaag
     5401 ctgatcgcac gcaaaaaaga ttgggacccc aagaaatacg gcggattcga ttctcctaca
     5461 gtcgcttaca gtgtactggt tgtggccaaa gtggagaaag ggaagtctaa aaaactcaaa
     5521 agcgtcaagg aactgctggg catcacaatc atggagcgat caagcttcga aaaaaacccc
     5581 atcgactttc tcgaggcgaa aggatataaa gaggtcaaaa aagacctcat cattaagctt
     5641 cccaagtact ctctctttga gcttgaaaac ggccggaaac gaatgctcgc tagtgcgggc
     5701 gagctgcaga aaggtaacga gctggcactg ccctctaaat acgttaattt cttgtatctg
     5761 gccagccact atgaaaagct caaagggtct cccgaagata atgagcagaa gcagctgttc
     5821 gtggaacaac acaaacacta ccttgatgag atcatcgagc aaataagcga attctccaaa
     5881 agagtgatcc tcgccgacgc taacctcgat aaggtgcttt ctgcttacaa taagcacagg
     5941 gataagccca tcagggagca ggcagaaaac attatccact tgtttactct gaccaacttg
     6001 ggcgcgcctg cagccttcaa gtacttcgac accaccatag acagaaagcg gtacacctct
     6061 acaaaggagg tcctggacgc cacactgatt catcagtcaa ttacggggct ctatgaaaca
     6121 agaatcgacc tctctcagct cggtggagac tctggtggtt caactaattt atctgatatt
     6181 attgaaaaag aaacaggtaa acaattagtt attcaagaat caattttaat gttgccagaa
     6241 gaagttgaag aagttattgg taataaacca gaatctgata ttttagttca tactgcttat
     6301 gatgaatcta ctgatgaaaa tgttatgtta ttgacttcag atgctccaga atataaacca
     6361 tgggctttag ttattcaaga ttctaatggt gaaaataaaa ttaaaatgtt gtctggtggt
     6421 tcacccaaga agaagaggaa ggtgtgatct cttctcgagt catgtaatta gttatgtcac
     6481 gcttacattc acgccctccc cccacatccg ctctaaccga aaaggaagga gttagacaac
     6541 ctgaagtcta ggtccctatt tattttttta tagttatgtt agtattaaga acgttattta
     6601 tatttcaaat ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac attatactga
     6661 aaaccttgct tgagaaggtt ttgggacgct cgaaggcttt aatttgcggc cggtacccaa
     6721 ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga
     6781 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     6841 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     6901 tggcgaatgg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     6961 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     7021 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     7081 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     7141 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     7201 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     7261 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     7321 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttcctg atgcggtatt
     7381 ttctccttac gcatctgtgc ggtatttcac accgcatatc gaccggtcga ggagaacttc
     7441 tagtatatct acatacctaa tattattgcc ttattaaaaa tggaatccca acaattacat
     7501 caaaatccac attctcttca aaatcaattg tcctgtactt ccttgttcat gtgtgttcaa
     7561 aaacgttata tttataggat aattatactc tatttctcaa caagtaattg gttgtttggc
     7621 cgagcggtct aaggcgcctg attcaagaaa tatcttgacc gcagttaact gtgggaatac
     7681 tcaggtatcg taagatgcaa gagttcgaat ctcttagcaa ccattatttt tttcctcaac
     7741 ataacgagaa cacacagggg cgctatcgca cagaatcaaa ttcgatgact ggaaattttt
     7801 tgttaatttc agaggtcgcc tgacgcatat acctttttca actgaaaaat tgggagaaaa
     7861 aggaaaggtg agagcgccgg aaccggcttt tcatatagaa tagagaagcg ttcatgacta
     7921 aatgcttgca tcacaatact tgaagttgac aatattattt aaggacctat tgttttttcc
     7981 aataggtggt tagcaatcgt cttactttct aacttttctt accttttaca tttcagcaat
     8041 atatatatat atatttcaag gatataccat tctaatgtct gcccctaaga agatcgtcgt
     8101 tttgccaggt gaccacgttg gtcaagaaat cacagccgaa gccattaagg ttcttaaagc
     8161 tatttctgat gttcgttcca atgtcaagtt cgatttcgaa aatcatttaa ttggtggtgc
     8221 tgctatcgat gctacaggtg ttccacttcc agatgaggcg ctggaagcct ccaagaaggc
     8281 tgatgccgtt ttgttaggtg ctgtgggtgg tcctaaatgg ggtaccggta gtgttagacc
     8341 tgaacaaggt ttactaaaaa tccgtaaaga acttcaattg tacgccaact taagaccatg
     8401 taactttgca tccgactctc ttttagactt atctccaatc aagccacaat ttgctaaagg
     8461 tactgacttc gttgttgtca gagaattagt gggaggtatt tactttggta agagaaagga
     8521 agacgatggt gatggtgtcg cttgggatag tgaacaatac accgttccag aagtgcaaag
     8581 aatcacaaga atggccgctt tcatggccct acaacatgag ccaccattgc ctatttggtc
     8641 cttggataaa gctaatgttt tggcctcttc aagattatgg agaaaaactg tggaggaaac
     8701 catcaagaac gaattcccta cattgaaggt tcaacatcaa ttgattgatt ctgccgccat
     8761 gatcctagtt aagaacccaa cccacctaaa tggtattata atcaccagca acatgtttgg
     8821 tgatatcatc tccgatgaag cctccgttat cccaggttcc ttgggtttgt tgccatctgc
     8881 gtccttggcc tctttgccag acaagaacac cgcatttggt ttgtacgaac catgccacgg
     8941 ttctgctcca gatttgccaa agaataaggt caaccctatc gccactatct tgtctgctgc
     9001 aatgatgttg aaattgtcat tgaacttgcc tgaagaaggt aaggccattg aagatgcagt
     9061 taaaaaggtt ttggatgcag gtatcagaac tggtgattta ggtggttcca acagtaccac
     9121 cgaagtcggt gatgctgtcg ccgaagaagt taagaaaatc cttgcttaaa aagattctct
     9181 ttttttatga tatttgtaca taaactttat aaatgaaatt cataatagaa acgacacgaa
     9241 attacaaaat ggaatatgtt catagggtag acgaaactat atacgcaatc tacatacatt
     9301 tatcaagaag gagaaaaagg aggatgtaaa ggaatacagg taagcaaatt gatactaatg
     9361 gctcaacgtg ataaggaaaa agaattgcac tttaacatta atattgacaa ggaggagggc
     9421 accacacaaa aagttaggtg taacagaaaa tcatgaaact atgattccta atttatatat
     9481 tggaggattt tctctaaaaa aaaaaaaata caacaaataa aaaacactca atgacctgac
     9541 catttgatgg agtttaagtc aataccttct tgaaccattt cccataatgg tgaaagttcc
     9601 ctcaagaatt ttactctgtc agaaacggcc ttaacgacgt agtcgatatg gtgcactctc
     9661 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     9721 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     9781 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     9841 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttaggac
     9901 ggatcgcttg cctgtaactt acacgcgcct cgtatctttt aatgatggaa taatttggga
     9961 atttactctg tgtttattta tttttatgtt ttgtatttgg attttagaaa gtaaataaag
    10021 aaggtagaag agttacggaa tgaagaaaaa aaaataaaca aaggtttaaa aaatttcaac
    10081 aaaaagcgta ctttacatat atatttatta gacaagaaaa gcagattaaa tagatataca
    10141 ttcgattaac gataagtaaa atgtaaaatc acaggatttt cgtgtgtggt cttctacaca
    10201 gacaagatga aacaattcgg cattaatacc tgagagcagg aagagcaaga taaaaggtag
    10261 tatttgttgg cgatccccct agagtctttt acatcttcgg aaaacaaaaa ctattttttc
    10321 tttaatttct ttttttactt tctattttta atttatatat ttatattaaa aaatttaaat
    10381 tataattatt tttatagcac gtgatgaaaa ggacccaggt ggcacttttc ggggaaatgt
    10441 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
    10501 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
    10561 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
    10621 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
    10681 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
    10741 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
    10801 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
    10861 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
    10921 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
    10981 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
    11041 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
    11101 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
    11161 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
    11221 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
    11281 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
    11341 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
    11401 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
    11461 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
    11521 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttc
//
