LOCUS       sequence-305805-       11613 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses nCDA1-Delta-190-BE3 in yeast cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11613)
  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 11613)
  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: pJT90_GalL_nCDA1-Delta-190-BE3
FEATURES             Location/Qualifiers
     source          1..11613
                     /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(2083..2109,2110..2112,2113..6186)
                     /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             2083..6186
                     /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             6199..6447
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6460..6480
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      6496..6743
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(6512..6530)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(6761..6780)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6762..6780)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6790..6807)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6790..6806)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6799..6821)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6947..7402
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7034..7053)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7244..7265
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7703..8110
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             8111..9205
                     /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(9699..9718)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9818..9840
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(9878..9896)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    9938..10441
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        10478..10582
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10583..10651,10652..11443)
                     /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(10801..10820)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     1021 tcactaaagg gaacaaaagc tggagctcgt gacagccctc cgaaggaaga ctctcctccg
     1081 tgcgtcctcg tcttcaccgg tcgcgttcct gaaacgcaga tgtgcctcgc gccgcactgc
     1141 tccgaacaat aaagattcta caatactagc ttttatggtt atgaagagga aaaattggca
     1201 gtaacctggc cccacaaacc ttcaaatgaa cgaatcaaat taacaaccat aggatgataa
     1261 tgcgattagt tttttagcct tatttctggg gtaattaatc agcgaagcga tgatttttga
     1321 tctattaacg gatatataaa tgcaaaaact gcataaccac tttaactaat actttcaaca
     1381 ttttcggttt gtattacttc ttattcaaat gtaataaaag tatcaacaaa aaattgttaa
     1441 tatacctcta tactttaacg tcaaggagaa aaaaccccgg attctagaac tagtggatcc
     1501 cccgggaaaa aaatgaccga cgctgagtac gtgagaatcc atgagaagtt ggacatctac
     1561 acgtttaaga aacagttttt caacaacaaa aaatccgtgt cgcatagatg ctacgttctc
     1621 tttgaattaa aacgacgggg tgaacgtaga gcgtgttttt ggggctatgc tgtgaataaa
     1681 ccacagagcg ggacagaacg tggcattcac gccgaaatct ttagcattag aaaagtcgaa
     1741 gaatacctgc gcgacaaccc cggacaattc acgataaatt ggtactcatc ctggagtcct
     1801 tgtgcagatt gcgctgaaaa gatcttagaa tggtataacc aggagctgcg ggggaacggc
     1861 cacactttga aaatctgggc ttgcaaactc tattacgaga aaaatgcgag gaatcaaatt
     1921 gggctgtgga acctcagaga taacggggtt gggttgaatg taatggtaag tgaacactac
     1981 caatgttgca ggaaaatatt catccaatcg tcgcacaatc aattgaatga gaatagatgg
     2041 cttgagaaga ctttgaagcg agctgaaaaa cgacggagcg agatggacaa gaagtactcc
     2101 attgggctcg ctatcggcac aaacagcgtc ggttgggccg tcattacgga cgagtacaag
     2161 gtgccgagca aaaaattcaa agttctgggc aataccgatc gccacagcat aaagaagaac
     2221 ctcattggcg ccctcctgtt cgactccggg gagacggccg aagccacgcg gctcaaaaga
     2281 acagcacggc gcagatatac ccgcagaaag aatcggatct gctacctgca ggagatcttt
     2341 agtaatgaga tggctaaggt ggatgactct ttcttccata ggctggagga gtcctttttg
     2401 gtggaggagg ataaaaagca cgagcgccac ccaatctttg gcaatatcgt ggacgaggtg
     2461 gcgtaccatg aaaagtaccc aaccatatat catctgagga agaagcttgt agacagtact
     2521 gataaggctg acttgcggtt gatctatctc gcgctggcgc atatgatcaa atttcgggga
     2581 cacttcctca tcgaggggga cctgaaccca gacaacagcg atgtcgacaa actctttatc
     2641 caactggttc agacttacaa tcagcttttc gaagagaacc cgatcaacgc atccggagtt
     2701 gacgccaaag caatcctgag cgctaggctg tccaaatccc ggcggctcga aaacctcatc
     2761 gcacagctcc ctggggagaa gaagaacggc ctgtttggta atcttatcgc cctgtcactc
     2821 gggctgaccc ccaactttaa atctaacttc gacctggccg aagatgccaa gcttcaactg
     2881 agcaaagaca cctacgatga tgatctcgac aatctgctgg cccagatcgg cgaccagtac
     2941 gcagaccttt ttttggcggc aaagaacctg tcagacgcca ttctgctgag tgatattctg
     3001 cgagtgaaca cggagatcac caaagctccg ctgagcgcta gtatgatcaa gcgctatgat
     3061 gagcaccacc aagacttgac tttgctgaag gcccttgtca gacagcaact gcctgagaag
     3121 tacaaggaaa ttttcttcga tcagtctaaa aatggctacg ccggatacat tgacggcgga
     3181 gcaagccagg aggaatttta caaatttatt aagcccatct tggaaaaaat ggacggcacc
     3241 gaggagctgc tggtaaagct taacagagaa gatctgttgc gcaaacagcg cactttcgac
     3301 aatggaagca tcccccacca gattcacctg ggcgaactgc acgctatcct caggcggcaa
     3361 gaggatttct accccttttt gaaagataac agggaaaaga ttgagaaaat cctcacattt
     3421 cggataccct actatgtagg ccccctcgcc cggggaaatt ccagattcgc gtggatgact
     3481 cgcaaatcag aagagaccat cactccctgg aacttcgagg aagtcgtgga taagggggcc
     3541 tctgcccagt ccttcatcga aaggatgact aactttgata aaaatctgcc taacgaaaag
     3601 gtgcttccta aacactctct gctgtacgag tacttcacag tttataacga gctcaccaag
     3661 gtcaaatacg tcacagaagg gatgagaaag ccagcattcc tgtctggaga gcagaagaaa
     3721 gctatcgtgg acctcctctt caagacgaac cggaaagtta ccgtgaaaca gctcaaagaa
     3781 gactatttca aaaagattga atgtttcgac tctgttgaaa tcagcggagt ggaggatcgc
     3841 ttcaacgcat ccctgggaac gtatcacgat ctcctgaaaa tcattaaaga caaggacttc
     3901 ctggacaatg aggagaacga ggacattctt gaggacattg tcctcaccct tacgttgttt
     3961 gaagataggg agatgattga agaacgcttg aaaacttacg ctcatctctt cgacgacaaa
     4021 gtcatgaaac agctcaagag gcgccgatat acaggatggg ggcggctgtc aagaaaactg
     4081 atcaatggga tccgagacaa gcagagtgga aagacaatcc tggattttct taagtccgat
     4141 ggatttgcca accggaactt catgcagttg atccatgatg actctctcac ctttaaggag
     4201 gacatccaga aagcacaagt ttctggccag ggggacagtc ttcacgagca catcgctaat
     4261 cttgcaggta gcccagctat caaaaaggga atactgcaga ccgttaaggt cgtggatgaa
     4321 ctcgtcaaag taatgggaag gcataagccc gagaatatcg ttatcgagat ggcccgagag
     4381 aaccaaacta cccagaaggg acagaagaac agtagggaaa ggatgaagag gattgaagag
     4441 ggtataaaag aactggggtc ccaaatcctt aaggaacacc cagttgaaaa cacccagctt
     4501 cagaatgaga agctctacct gtactacctg cagaacggca gggacatgta cgtggatcag
     4561 gaactggaca tcaatcggct ctccgactac gacgtggatc atatcgtgcc ccagtctttt
     4621 ctcaaagatg attctattga taataaagtg ttgacaagat ccgataaaaa tagagggaag
     4681 agtgataacg tcccctcaga agaagttgtc aagaaaatga aaaattattg gcggcagctg
     4741 ctgaacgcca aactgatcac acaacggaag ttcgataatc tgactaaggc tgaacgaggt
     4801 ggcctgtctg agttggataa agccggcttc atcaaaaggc agcttgttga gacacgccag
     4861 atcaccaagc acgtggccca aattctcgat tcacgcatga acaccaagta cgatgaaaat
     4921 gacaaactga ttcgagaggt gaaagttatt actctgaagt ctaagctggt ctcagatttc
     4981 agaaaggact ttcagtttta taaggtgaga gagatcaaca attaccacca tgcgcatgat
     5041 gcctacctga atgcagtggt aggcactgca cttatcaaaa aatatcccaa gcttgaatct
     5101 gaatttgttt acggagacta taaagtgtac gatgttagga aaatgatcgc aaagtctgag
     5161 caggaaatag gcaaggccac cgctaagtac ttcttttaca gcaatattat gaattttttc
     5221 aagaccgaga ttacactggc caatggagag attcggaagc gaccacttat cgaaacaaac
     5281 ggagaaacag gagaaatcgt gtgggacaag ggtagggatt tcgcgacagt ccggaaggtc
     5341 ctgtccatgc cgcaggtgaa catcgttaaa aagaccgaag tacagaccgg aggcttctcc
     5401 aaggaaagta tcctcccgaa aaggaacagc gacaagctga tcgcacgcaa aaaagattgg
     5461 gaccccaaga aatacggcgg attcgattct cctacagtcg cttacagtgt actggttgtg
     5521 gccaaagtgg agaaagggaa gtctaaaaaa ctcaaaagcg tcaaggaact gctgggcatc
     5581 acaatcatgg agcgatcaag cttcgaaaaa aaccccatcg actttctcga ggcgaaagga
     5641 tataaagagg tcaaaaaaga cctcatcatt aagcttccca agtactctct ctttgagctt
     5701 gaaaacggcc ggaaacgaat gctcgctagt gcgggcgagc tgcagaaagg taacgagctg
     5761 gcactgccct ctaaatacgt taatttcttg tatctggcca gccactatga aaagctcaaa
     5821 gggtctcccg aagataatga gcagaagcag ctgttcgtgg aacaacacaa acactacctt
     5881 gatgagatca tcgagcaaat aagcgaattc tccaaaagag tgatcctcgc cgacgctaac
     5941 ctcgataagg tgctttctgc ttacaataag cacagggata agcccatcag ggagcaggca
     6001 gaaaacatta tccacttgtt tactctgacc aacttgggcg cgcctgcagc cttcaagtac
     6061 ttcgacacca ccatagacag aaagcggtac acctctacaa aggaggtcct ggacgccaca
     6121 ctgattcatc agtcaattac ggggctctat gaaacaagaa tcgacctctc tcagctcggt
     6181 ggagactctg gtggttcaac taatttatct gatattattg aaaaagaaac aggtaaacaa
     6241 ttagttattc aagaatcaat tttaatgttg ccagaagaag ttgaagaagt tattggtaat
     6301 aaaccagaat ctgatatttt agttcatact gcttatgatg aatctactga tgaaaatgtt
     6361 atgttattga cttcagatgc tccagaatat aaaccatggg ctttagttat tcaagattct
     6421 aatggtgaaa ataaaattaa aatgttgtct ggtggttcac ccaagaagaa gaggaaggtg
     6481 tgatctcttc tcgagtcatg taattagtta tgtcacgctt acattcacgc cctcccccca
     6541 catccgctct aaccgaaaag gaaggagtta gacaacctga agtctaggtc cctatttatt
     6601 tttttatagt tatgttagta ttaagaacgt tatttatatt tcaaattttt cttttttttc
     6661 tgtacagacg cgtgtacgca tgtaacatta tactgaaaac cttgcttgag aaggttttgg
     6721 gacgctcgaa ggctttaatt tgcggccggt acccaattcg ccctatagtg agtcgtatta
     6781 cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     6841 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     6901 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggacgc gccctgtagc
     6961 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     7021 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     7081 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     7141 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     7201 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     7261 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     7321 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     7381 aaaatattaa cgcttacaat ttcctgatgc ggtattttct ccttacgcat ctgtgcggta
     7441 tttcacaccg catatcgacc ggtcgaggag aacttctagt atatctacat acctaatatt
     7501 attgccttat taaaaatgga atcccaacaa ttacatcaaa atccacattc tcttcaaaat
     7561 caattgtcct gtacttcctt gttcatgtgt gttcaaaaac gttatattta taggataatt
     7621 atactctatt tctcaacaag taattggttg tttggccgag cggtctaagg cgcctgattc
     7681 aagaaatatc ttgaccgcag ttaactgtgg gaatactcag gtatcgtaag atgcaagagt
     7741 tcgaatctct tagcaaccat tatttttttc ctcaacataa cgagaacaca caggggcgct
     7801 atcgcacaga atcaaattcg atgactggaa attttttgtt aatttcagag gtcgcctgac
     7861 gcatatacct ttttcaactg aaaaattggg agaaaaagga aaggtgagag cgccggaacc
     7921 ggcttttcat atagaataga gaagcgttca tgactaaatg cttgcatcac aatacttgaa
     7981 gttgacaata ttatttaagg acctattgtt ttttccaata ggtggttagc aatcgtctta
     8041 ctttctaact tttcttacct tttacatttc agcaatatat atatatatat ttcaaggata
     8101 taccattcta atgtctgccc ctaagaagat cgtcgttttg ccaggtgacc acgttggtca
     8161 agaaatcaca gccgaagcca ttaaggttct taaagctatt tctgatgttc gttccaatgt
     8221 caagttcgat ttcgaaaatc atttaattgg tggtgctgct atcgatgcta caggtgttcc
     8281 acttccagat gaggcgctgg aagcctccaa gaaggctgat gccgttttgt taggtgctgt
     8341 gggtggtcct aaatggggta ccggtagtgt tagacctgaa caaggtttac taaaaatccg
     8401 taaagaactt caattgtacg ccaacttaag accatgtaac tttgcatccg actctctttt
     8461 agacttatct ccaatcaagc cacaatttgc taaaggtact gacttcgttg ttgtcagaga
     8521 attagtggga ggtatttact ttggtaagag aaaggaagac gatggtgatg gtgtcgcttg
     8581 ggatagtgaa caatacaccg ttccagaagt gcaaagaatc acaagaatgg ccgctttcat
     8641 ggccctacaa catgagccac cattgcctat ttggtccttg gataaagcta atgttttggc
     8701 ctcttcaaga ttatggagaa aaactgtgga ggaaaccatc aagaacgaat tccctacatt
     8761 gaaggttcaa catcaattga ttgattctgc cgccatgatc ctagttaaga acccaaccca
     8821 cctaaatggt attataatca ccagcaacat gtttggtgat atcatctccg atgaagcctc
     8881 cgttatccca ggttccttgg gtttgttgcc atctgcgtcc ttggcctctt tgccagacaa
     8941 gaacaccgca tttggtttgt acgaaccatg ccacggttct gctccagatt tgccaaagaa
     9001 taaggtcaac cctatcgcca ctatcttgtc tgctgcaatg atgttgaaat tgtcattgaa
     9061 cttgcctgaa gaaggtaagg ccattgaaga tgcagttaaa aaggttttgg atgcaggtat
     9121 cagaactggt gatttaggtg gttccaacag taccaccgaa gtcggtgatg ctgtcgccga
     9181 agaagttaag aaaatccttg cttaaaaaga ttctcttttt ttatgatatt tgtacataaa
     9241 ctttataaat gaaattcata atagaaacga cacgaaatta caaaatggaa tatgttcata
     9301 gggtagacga aactatatac gcaatctaca tacatttatc aagaaggaga aaaaggagga
     9361 tgtaaaggaa tacaggtaag caaattgata ctaatggctc aacgtgataa ggaaaaagaa
     9421 ttgcacttta acattaatat tgacaaggag gagggcacca cacaaaaagt taggtgtaac
     9481 agaaaatcat gaaactatga ttcctaattt atatattgga ggattttctc taaaaaaaaa
     9541 aaaatacaac aaataaaaaa cactcaatga cctgaccatt tgatggagtt taagtcaata
     9601 ccttcttgaa ccatttccca taatggtgaa agttccctca agaattttac tctgtcagaa
     9661 acggccttaa cgacgtagtc gatatggtgc actctcagta caatctgctc tgatgccgca
     9721 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     9781 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     9841 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
     9901 taggttaatg tcatgataat aatggtttct taggacggat cgcttgcctg taacttacac
     9961 gcgcctcgta tcttttaatg atggaataat ttgggaattt actctgtgtt tatttatttt
    10021 tatgttttgt atttggattt tagaaagtaa ataaagaagg tagaagagtt acggaatgaa
    10081 gaaaaaaaaa taaacaaagg tttaaaaaat ttcaacaaaa agcgtacttt acatatatat
    10141 ttattagaca agaaaagcag attaaataga tatacattcg attaacgata agtaaaatgt
    10201 aaaatcacag gattttcgtg tgtggtcttc tacacagaca agatgaaaca attcggcatt
    10261 aatacctgag agcaggaaga gcaagataaa aggtagtatt tgttggcgat ccccctagag
    10321 tcttttacat cttcggaaaa caaaaactat tttttcttta atttcttttt ttactttcta
    10381 tttttaattt atatatttat attaaaaaat ttaaattata attattttta tagcacgtga
    10441 tgaaaaggac ccaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
    10501 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
    10561 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
    10621 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
    10681 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
    10741 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
    10801 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
    10861 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
    10921 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
    10981 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
    11041 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
    11101 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
    11161 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
    11221 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
    11281 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
    11341 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
    11401 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
    11461 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
    11521 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
    11581 gtcagacccc gtagaaaaga tcaaaggatc ttc
//
