LOCUS       sequence-273643-       11313 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mTN3-GGS6-dCas9 (iCas9) in yeast. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11313)
  AUTHORS   Standage-Beier K, Brookhouser N, Balachandran P, Zhang Q, Brafman 
            DA, Wang X
  TITLE     RNA-Guided Recombinase-Cas9 Fusion Targets Genomic DNA Deletion and 
            Integration.
  JOURNAL   CRISPR J. 2019 Aug;2:209-222. doi: 10.1089/crispr.2019.0013.
  PUBMED    31436506
REFERENCE   2  (bases 1 to 11313)
  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: "CRISPR J.";
            date: "2019-08"; volume: "2:209-222. doi"; pages: " 
            10.1089/crispr.2019.0013"
COMMENT     Sequence Label: p415:iCas9
FEATURES             Location/Qualifiers
     source          1..11313
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        79..183
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(184..252,253..1044)
                     /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(402..421)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1215..1803
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1704..1723
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1957..1974
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2091..2112
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2127..2132,2133..2150,2151..2157)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2165..2181
                     /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     2170..2192
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2189..2205
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2189..2205
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2224..2244
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2226..2244
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     2653..2676
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     2697..2713
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2697..2713
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             2730..2750
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(3258..3284,3285..3287,3288..5771,5772..5777,5778..5843,
                     5844..5846,5847..7361)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=Cas9m4
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the mutations D10A, D839A, 
                     H840A, and N863A (Mali et al., 2013)"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             join(3258..3284,3285..3287,3288..5774,5775..5777,
                     5778..7361)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             3258..7361
                     /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
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             7374..7394
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      7410..7657
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(7426..7444)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(7675..7694)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7676..7694)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7704..7721)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7704..7720)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7713..7735)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7861..8316
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7948..7967)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8158..8179
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8617..9024
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             9025..10119
                     /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(10613..10632)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     10732..10754
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(10792..10810)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    join(10852..11313,1..42)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
ORIGIN
        1 tattaaaaaa tttaaattat aattattttt atagcacgtg atgaaaagga cccaggtggc
       61 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
      121 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
      181 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
      241 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
      301 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
      361 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
      421 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
      481 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
      541 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
      601 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
      661 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
      721 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
      781 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
      841 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
      901 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
      961 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     1021 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     1081 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     1141 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     1201 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     1261 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     1321 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     1381 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     1441 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     1501 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     1561 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     1621 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     1681 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     1741 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     1801 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     1861 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     1921 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     1981 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     2041 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     2101 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     2161 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     2221 cgcgcaatta accctcacta aagggaacaa aagctggagc tcgtgacagc cctccgaagg
     2281 aagactctcc tccgtgcgtc ctcgtcttca ccggtcgcgt tcctgaaacg cagatgtgcc
     2341 tcgcgccgca ctgctccgaa caataaagat tctacaatac tagcttttat ggttatgaag
     2401 aggaaaaatt ggcagtaacc tggccccaca aaccttcaaa tgaacgaatc aaattaacaa
     2461 ccataggatg ataatgcgat tagtttttta gccttatttc tggggtaatt aatcagcgaa
     2521 gcgatgattt ttgatctatt aacggatata taaatgcaaa aactgcataa ccactttaac
     2581 taatactttc aacattttcg gtttgtatta cttcttattc aaatgtaata aaagtatcaa
     2641 caaaaaattg ttaatatacc tctatacttt aacgtcaagg agaaaaaacc ccggattcta
     2701 gaactagtgg atcccccggg aaaaaaatgc ccaaaaagaa gaggaaagta ggtggcagta
     2761 tgaggatatt tggatacgca cgagttagta catctcaaca gtctctggac atacagataa
     2821 gggcattgaa ggatgctgga gtgaaagcta atcggatctt taccgataag gctagtggca
     2881 gctctacaga cagagagggc ttggatctgc tgagaatgaa ggttgaggaa ggggatgtga
     2941 tactcgtcaa gaaacttgac agactctcac gagacactgc cgatatgatc caactgatta
     3001 aggaatttga cgcccaaggt gtagccgtaa gatttattga tgacggtatc agcaccgacg
     3061 gctatatggg ccagatggtc gtcactatac tgagcgctgt cgcacaagcg gagagaagac
     3121 gaatcctgca acggacaaat gaaggtcgcc aagaagcaaa gctcaaagga attaagttcg
     3181 ggcgcaggag gactgtagac agaggtggtt caggaggaag tggaggttca gggggaagtg
     3241 gtggctccgg tgggtctatg gacaagaagt actccattgg gctcgccatc ggcacaaaca
     3301 gcgtaggttg ggccgtcatt acggacgagt acaaggtgcc gagcaaaaaa ttcaaagttc
     3361 tgggcaatac cgatcgccac agcataaaga agaacctcat tggcgccctc ctgttcgact
     3421 ccggggagac ggccgaagcc acgcggctca aaagaacagc acggcgcaga tatacccgca
     3481 gaaagaatcg gatctgctac ctgcaggaga tctttagtaa tgagatggct aaggtggatg
     3541 actctttctt ccataggctg gaggagtcct ttttggtgga ggaggataaa aagcacgagc
     3601 gccacccaat ctttggcaat atcgtggacg aggtggcgta ccatgaaaag tacccaacca
     3661 tatatcatct gaggaagaag cttgtagaca gtactgataa ggctgacttg cggttgatct
     3721 atctcgcgct ggcgcatatg atcaaatttc ggggacactt cctcatcgag ggggacctga
     3781 acccagacaa cagcgatgtc gacaaactct ttatccaact ggttcagact tacaatcagc
     3841 ttttcgaaga gaacccgatc aacgcatccg gagttgacgc caaagcaatc ctgagcgcta
     3901 ggctgtccaa atcccggcgg ctcgaaaacc tcatcgcaca gctccctggg gagaagaaga
     3961 acggcctgtt tggtaatctt atcgccctgt cactcgggct gacccccaac tttaaatcta
     4021 acttcgacct ggccgaagat gccaagcttc aactgagcaa agacacctac gatgatgatc
     4081 tcgacaatct gctggcccag atcggcgacc agtacgcaga cctttttttg gcggcaaaga
     4141 acctgtcaga cgccattctg ctgagtgata ttctgcgagt gaacacggag atcaccaaag
     4201 ctccgctgag cgctagtatg atcaagcgct atgatgagca ccaccaagac ttgactttgc
     4261 tgaaggccct tgtcagacag caactgcctg agaagtacaa ggaaattttc ttcgatcagt
     4321 ctaaaaatgg ctacgccgga tacattgacg gcggagcaag ccaggaggaa ttttacaaat
     4381 ttattaagcc catcttggaa aaaatggacg gcaccgagga gctgctggta aagcttaaca
     4441 gagaagatct gttgcgcaaa cagcgcactt tcgacaatgg aagcatcccc caccagattc
     4501 acctgggcga actgcacgct atcctcaggc ggcaagagga tttctacccc tttttgaaag
     4561 ataacaggga aaagattgag aaaatcctca catttcggat accctactat gtaggccccc
     4621 tcgcccgggg aaattccaga ttcgcgtgga tgactcgcaa atcagaagag accatcactc
     4681 cctggaactt cgaggaagtc gtggataagg gggcctctgc ccagtccttc atcgaaagga
     4741 tgactaactt tgataaaaat ctgcctaacg aaaaggtgct tcctaaacac tctctgctgt
     4801 acgagtactt cacagtttat aacgagctca ccaaggtcaa atacgtcaca gaagggatga
     4861 gaaagccagc attcctgtct ggagagcaga agaaagctat cgtggacctc ctcttcaaga
     4921 cgaaccggaa agttaccgtg aaacagctca aagaagacta tttcaaaaag attgaatgtt
     4981 tcgactctgt tgaaatcagc ggagtggagg atcgcttcaa cgcatccctg ggaacgtatc
     5041 acgatctcct gaaaatcatt aaagacaagg acttcctgga caatgaggag aacgaggaca
     5101 ttcttgagga cattgtcctc acccttacgt tgtttgaaga tagggagatg attgaagaac
     5161 gcttgaaaac ttacgctcat ctcttcgacg acaaagtcat gaaacagctc aagaggcgcc
     5221 gatatacagg atgggggcgg ctgtcaagaa aactgatcaa tgggatccga gacaagcaga
     5281 gtggaaagac aatcctggat tttcttaagt ccgatggatt tgccaaccgg aacttcatgc
     5341 agttgatcca tgatgactct ctcaccttta aggaggacat ccagaaagca caagtttctg
     5401 gccaggggga cagtcttcac gagcacatcg ctaatcttgc aggtagccca gctatcaaaa
     5461 agggaatact gcagaccgtt aaggtcgtgg atgaactcgt caaagtaatg ggaaggcata
     5521 agcccgagaa tatcgttatc gagatggccc gagagaacca aactacccag aagggacaga
     5581 agaacagtag ggaaaggatg aagaggattg aagagggtat aaaagaactg gggtcccaaa
     5641 tccttaagga acacccagtt gaaaacaccc agcttcagaa tgagaagctc tacctgtact
     5701 acctgcagaa cggcagggac atgtacgtgg atcaggaact ggacatcaat cggctctccg
     5761 actacgacgt ggatgccatc gtgccccagt cttttctcaa agatgattct attgataata
     5821 aagtgttgac aagatccgat aaaaatagag ggaagagtga taacgtcccc tcagaagaag
     5881 ttgtcaagaa aatgaaaaat tattggcggc agctgctgaa cgccaaactg atcacacaac
     5941 ggaagttcga taatctgact aaggctgaac gaggtggcct gtctgagttg gataaagccg
     6001 gcttcatcaa aaggcagctt gttgagacac gccagatcac caagcacgtg gcccaaattc
     6061 tcgattcacg catgaacacc aagtacgatg aaaatgacaa actgattcga gaggtgaaag
     6121 ttattactct gaagtctaag ctggtctcag atttcagaaa ggactttcag ttttataagg
     6181 tgagagagat caacaattac caccatgcgc atgatgccta cctgaatgca gtggtaggca
     6241 ctgcacttat caaaaaatat cccaagcttg aatctgaatt tgtttacgga gactataaag
     6301 tgtacgatgt taggaaaatg atcgcaaagt ctgagcagga aataggcaag gccaccgcta
     6361 agtacttctt ttacagcaat attatgaatt ttttcaagac cgagattaca ctggccaatg
     6421 gagagattcg gaagcgacca cttatcgaaa caaacggaga aacaggagaa atcgtgtggg
     6481 acaagggtag ggatttcgcg acagtccgga aggtcctgtc catgccgcag gtgaacatcg
     6541 ttaaaaagac cgaagtacag accggaggct tctccaagga aagtatcctc ccgaaaagga
     6601 acagcgacaa gctgatcgca cgcaaaaaag attgggaccc caagaaatac ggcggattcg
     6661 attctcctac agtcgcttac agtgtactgg ttgtggccaa agtggagaaa gggaagtcta
     6721 aaaaactcaa aagcgtcaag gaactgctgg gcatcacaat catggagcga tcaagcttcg
     6781 aaaaaaaccc catcgacttt ctcgaggcga aaggatataa agaggtcaaa aaagacctca
     6841 tcattaagct tcccaagtac tctctctttg agcttgaaaa cggccggaaa cgaatgctcg
     6901 ctagtgcggg cgagctgcag aaaggtaacg agctggcact gccctctaaa tacgttaatt
     6961 tcttgtatct ggccagccac tatgaaaagc tcaaagggtc tcccgaagat aatgagcaga
     7021 agcagctgtt cgtggaacaa cacaaacact accttgatga gatcatcgag caaataagcg
     7081 aattctccaa aagagtgatc ctcgccgacg ctaacctcga taaggtgctt tctgcttaca
     7141 ataagcacag ggataagccc atcagggagc aggcagaaaa cattatccac ttgtttactc
     7201 tgaccaactt gggcgcgcct gcagccttca agtacttcga caccaccata gacagaaagc
     7261 ggtacacctc tacaaaggag gtcctggacg ccacactgat tcatcagtca attacggggc
     7321 tctatgaaac aagaatcgac ctctctcagc tcggtggaga cagcagggct gaccccaaga
     7381 agaagaggaa ggtgtgatct cttctcgagt catgtaatta gttatgtcac gcttacattc
     7441 acgccctccc cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta
     7501 ggtccctatt tattttttta tagttatgtt agtattaaga acgttattta tatttcaaat
     7561 ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac attatactga aaaccttgct
     7621 tgagaaggtt ttgggacgct cgaaggcttt aatttgcggc cggtacccaa ttcgccctat
     7681 agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     7741 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     7801 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     7861 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     7921 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     7981 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     8041 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     8101 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     8161 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     8221 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     8281 acgcgaattt taacaaaata ttaacgctta caatttcctg atgcggtatt ttctccttac
     8341 gcatctgtgc ggtatttcac accgcatatc gaccggtcga ggagaacttc tagtatatct
     8401 acatacctaa tattattgcc ttattaaaaa tggaatccca acaattacat caaaatccac
     8461 attctcttca aaatcaattg tcctgtactt ccttgttcat gtgtgttcaa aaacgttata
     8521 tttataggat aattatactc tatttctcaa caagtaattg gttgtttggc cgagcggtct
     8581 aaggcgcctg attcaagaaa tatcttgacc gcagttaact gtgggaatac tcaggtatcg
     8641 taagatgcaa gagttcgaat ctcttagcaa ccattatttt tttcctcaac ataacgagaa
     8701 cacacagggg cgctatcgca cagaatcaaa ttcgatgact ggaaattttt tgttaatttc
     8761 agaggtcgcc tgacgcatat acctttttca actgaaaaat tgggagaaaa aggaaaggtg
     8821 agagcgccgg aaccggcttt tcatatagaa tagagaagcg ttcatgacta aatgcttgca
     8881 tcacaatact tgaagttgac aatattattt aaggacctat tgttttttcc aataggtggt
     8941 tagcaatcgt cttactttct aacttttctt accttttaca tttcagcaat atatatatat
     9001 atatttcaag gatataccat tctaatgtct gcccctaaga agatcgtcgt tttgccaggt
     9061 gaccacgttg gtcaagaaat cacagccgaa gccattaagg ttcttaaagc tatttctgat
     9121 gttcgttcca atgtcaagtt cgatttcgaa aatcatttaa ttggtggtgc tgctatcgat
     9181 gctacaggtg ttccacttcc agatgaggcg ctggaagcct ccaagaaggc tgatgccgtt
     9241 ttgttaggtg ctgtgggtgg tcctaaatgg ggtaccggta gtgttagacc tgaacaaggt
     9301 ttactaaaaa tccgtaaaga acttcaattg tacgccaact taagaccatg taactttgca
     9361 tccgactctc ttttagactt atctccaatc aagccacaat ttgctaaagg tactgacttc
     9421 gttgttgtca gagaattagt gggaggtatt tactttggta agagaaagga agacgatggt
     9481 gatggtgtcg cttgggatag tgaacaatac accgttccag aagtgcaaag aatcacaaga
     9541 atggccgctt tcatggccct acaacatgag ccaccattgc ctatttggtc cttggataaa
     9601 gctaatgttt tggcctcttc aagattatgg agaaaaactg tggaggaaac catcaagaac
     9661 gaattcccta cattgaaggt tcaacatcaa ttgattgatt ctgccgccat gatcctagtt
     9721 aagaacccaa cccacctaaa tggtattata atcaccagca acatgtttgg tgatatcatc
     9781 tccgatgaag cctccgttat cccaggttcc ttgggtttgt tgccatctgc gtccttggcc
     9841 tctttgccag acaagaacac cgcatttggt ttgtacgaac catgccacgg ttctgctcca
     9901 gatttgccaa agaataaggt caaccctatc gccactatct tgtctgctgc aatgatgttg
     9961 aaattgtcat tgaacttgcc tgaagaaggt aaggccattg aagatgcagt taaaaaggtt
    10021 ttggatgcag gtatcagaac tggtgattta ggtggttcca acagtaccac cgaagtcggt
    10081 gatgctgtcg ccgaagaagt taagaaaatc cttgcttaaa aagattctct ttttttatga
    10141 tatttgtaca taaactttat aaatgaaatt cataatagaa acgacacgaa attacaaaat
    10201 ggaatatgtt catagggtag acgaaactat atacgcaatc tacatacatt tatcaagaag
    10261 gagaaaaagg aggatgtaaa ggaatacagg taagcaaatt gatactaatg gctcaacgtg
    10321 ataaggaaaa agaattgcac tttaacatta atattgacaa ggaggagggc accacacaaa
    10381 aagttaggtg taacagaaaa tcatgaaact atgattccta atttatatat tggaggattt
    10441 tctctaaaaa aaaaaaaata caacaaataa aaaacactca atgacctgac catttgatgg
    10501 agtttaagtc aataccttct tgaaccattt cccataatgg tgaaagttcc ctcaagaatt
    10561 ttactctgtc agaaacggcc ttaacgacgt agtcgatatg gtgcactctc agtacaatct
    10621 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
    10681 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
    10741 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
    10801 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttaggac ggatcgcttg
    10861 cctgtaactt acacgcgcct cgtatctttt aatgatggaa taatttggga atttactctg
    10921 tgtttattta tttttatgtt ttgtatttgg attttagaaa gtaaataaag aaggtagaag
    10981 agttacggaa tgaagaaaaa aaaataaaca aaggtttaaa aaatttcaac aaaaagcgta
    11041 ctttacatat atatttatta gacaagaaaa gcagattaaa tagatataca ttcgattaac
    11101 gataagtaaa atgtaaaatc acaggatttt cgtgtgtggt cttctacaca gacaagatga
    11161 aacaattcgg cattaatacc tgagagcagg aagagcaaga taaaaggtag tatttgttgg
    11221 cgatccccct agagtctttt acatcttcgg aaaacaaaaa ctattttttc tttaatttct
    11281 ttttttactt tctattttta atttatatat tta
//
