LOCUS       sequence-127063-       10203 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This plasmid contains a Cas9 Activator for yeast. The activator is 
            about 1.2-1.8 X more potent than just dCas9-VP64 fusion in yeast. It
            is on a single copy CEN/ARS plasmid with Ura marker, pRS416..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10203)
  AUTHORS   Naranjo S, Smith JD, Artieri CG, Zhang M, Zhou Y, Palmer ME, Fraser 
            HB
  TITLE     Dissecting the Genetic Basis of a Complex cis-Regulatory Adaptation.
  JOURNAL   PLoS Genet. 2015 Dec 29;11(12):e1005751. doi: 
            10.1371/journal.pgen.1005751. eCollection 2015 Dec.
  PUBMED    26713447
REFERENCE   2  (bases 1 to 10203)
  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: "PLoS 
            Genet."; date: "2015-12-29"; volume: "11(12):e1005751. doi"; pages: 
            " 10.1371/journal.pgen.1005751. eCollection 2015 Dec"
COMMENT     Sequence Label: pRS416-Gal4-dCas9-VP64
FEATURES             Location/Qualifiers
     source          1..10203
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        196..416
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             417..1220
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(1351..1806)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1488..1509)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1700..1719
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1936..1958
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1950..1967
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1951..1967
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1977..1996
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1977..1995
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(2014..2261)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     2227..2245
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(2280..2429)
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             complement(join(2478..3992,3993..3995,3996..4061,4062..4067,
                     4068..6551,6552..6554,6555..6581))
                     /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             complement(join(2478..4061,4062..4064,4065..6551,6552..6554,
                     6555..6581))
                     /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             complement(2478..6581)
                     /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"
     protein_bind    complement(6584..6608)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(6624..6965)
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="activation domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 activation domain
                     /translation="ANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
                     VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
                     EDTPPNPKKE"
     CDS             complement(6975..6995)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(7015..7413)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     complement(7431..7451)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7431..7449)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7470..7486)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7470..7486)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    7492..7508
                     /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)."
     promoter        complement(join(7516..7522,7523..7540,7541..7546))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(7699..7716)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7870..8458)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7950..7969)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8629..9420,9421..9489))
                     /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     9252..9271
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9490..9594)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    9631..10134
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     10176..10194
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataccac agcttttcaa ttcaattcat catttttttt ttattctttt ttttgatttc
      241 ggtttctttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg
      301 agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc
      361 cagtattctt aacccaactg cacagaacaa aaacctgcag gaaacgaaga taaatcatgt
      421 cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct gccaagctat
      481 ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt cgtaccacca
      541 aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta aaaacacatg
      601 tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag gcattatccg
      661 ccaagtacaa ttttttactc ttcgaagaca gaaaatttgc tgacattggt aatacagtca
      721 aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt acgaatgcac
      781 acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa gaagtaacaa
      841 aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc ctatctactg
      901 gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt gttatcggct
      961 ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg attatgacac
     1021 ccggtgtggg tttagatgac aagggagacg cattgggtca acagtataga accgtggatg
     1081 atgtggtctc tacaggatct gacattatta ttgttggaag aggactattt gcaaagggaa
     1141 gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca tatttgagaa
     1201 gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata ctaaactcac
     1261 aaattagagc ttcaatttaa ttatatcagt tattacccta tgcggtgtga aataccgcac
     1321 agatgcgtaa ggagaaaata ccgcatcagg aaattgtaaa cgttaatatt ttgttaaaat
     1381 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     1441 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     1501 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     1561 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     1621 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     1681 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     1741 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     1801 gcgcgtcgcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     1861 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     1921 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat
     1981 acgactcact atagggcgaa ttgggtaccg gccgcaaatt aaagccttcg agcgtcccaa
     2041 aaccttctca agcaaggttt tcagtataat gttacatgcg tacacgcgtc tgtacagaaa
     2101 aaaaagaaaa atttgaaata taaataacgt tcttaatact aacataacta taaaaaaata
     2161 aatagggacc tagacttcag gttgtctaac tccttccttt tcggttagag cggatgtggg
     2221 gggagggcgt gaatgtaagc gtgacataac taattacatg actcgagttc ttcctaggaa
     2281 agcatgtcta agtcgaaatc gtccagagcg tctgatccaa gcatatccag gtcaaaatca
     2341 tctaaggcgt cacttccaag catgtcgagg tcaaagtcat ctagcgcatc agaacccagc
     2401 atatccaagt cgaagtcgtc caaagcgtca gaacctccac cgtcgactcc agatctacca
     2461 gaaccgtcag ccctgctgtc tccaccgagc tgagagaggt cgattcttgt ttcatagagc
     2521 cccgtaattg actgatgaat cagtgtggcg tccaggacct cctttgtaga ggtgtaccgc
     2581 tttctgtcta tggtggtgtc gaagtacttg aaggctgcag gcgcgcccaa gttggtcaga
     2641 gtaaacaagt ggataatgtt ttctgcctgc tccctgatgg gcttatccct gtgcttattg
     2701 taagcagaaa gcaccttatc gaggttagcg tcggcgagga tcactctttt ggagaattcg
     2761 cttatttgct cgatgatctc atcaaggtag tgtttgtgtt gttccacgaa cagctgcttc
     2821 tgctcattat cttcgggaga ccctttgagc ttttcatagt ggctggccag atacaagaaa
     2881 ttaacgtatt tagagggcag tgccagctcg ttacctttct gcagctcgcc cgcactagcg
     2941 agcattcgtt tccggccgtt ttcaagctca aagagagagt acttgggaag cttaatgatg
     3001 aggtcttttt tgacctcttt atatcctttc gcctcgagaa agtcgatggg gtttttttcg
     3061 aagcttgatc gctccatgat tgtgatgccc agcagttcct tgacgctttt gagtttttta
     3121 gacttccctt tctccacttt ggccacaacc agtacactgt aagcgactgt aggagaatcg
     3181 aatccgccgt atttcttggg gtcccaatct tttttgcgtg cgatcagctt gtcgctgttc
     3241 cttttcggga ggatactttc cttggagaag cctccggtct gtacttcggt ctttttaacg
     3301 atgttcacct gcggcatgga caggaccttc cggactgtcg cgaaatccct acccttgtcc
     3361 cacacgattt ctcctgtttc tccgtttgtt tcgataagtg gtcgcttccg aatctctcca
     3421 ttggccagtg taatctcggt cttgaaaaaa ttcataatat tgctgtaaaa gaagtactta
     3481 gcggtggcct tgcctatttc ctgctcagac tttgcgatca ttttcctaac atcgtacact
     3541 ttatagtctc cgtaaacaaa ttcagattca agcttgggat attttttgat aagtgcagtg
     3601 cctaccactg cattcaggta ggcatcatgc gcatggtggt aattgttgat ctctctcacc
     3661 ttataaaact gaaagtcctt tctgaaatct gagaccagct tagacttcag agtaataact
     3721 ttcacctctc gaatcagttt gtcattttca tcgtacttgg tgttcatgcg tgaatcgaga
     3781 atttgggcca cgtgcttggt gatctggcgt gtctcaacaa gctgcctttt gatgaagccg
     3841 gctttatcca actcagacag gccacctcgt tcagccttag tcagattatc gaacttccgt
     3901 tgtgtgatca gtttggcgtt cagcagctgc cgccaataat ttttcatttt cttgacaact
     3961 tcttctgagg ggacgttatc actcttccct ctatttttat cggatcttgt caacacttta
     4021 ttatcaatag aatcatcttt gagaaaagac tggggcacga tagcatccac gtcgtagtcg
     4081 gagagccgat tgatgtccag ttcctgatcc acgtacatgt ccctgccgtt ctgcaggtag
     4141 tacaggtaga gcttctcatt ctgaagctgg gtgttttcaa ctgggtgttc cttaaggatt
     4201 tgggacccca gttcttttat accctcttca atcctcttca tcctttccct actgttcttc
     4261 tgtcccttct gggtagtttg gttctctcgg gccatctcga taacgatatt ctcgggctta
     4321 tgccttccca ttactttgac gagttcatcc acgaccttaa cggtctgcag tattcccttt
     4381 ttgatagctg ggctacctgc aagattagcg atgtgctcgt gaagactgtc cccctggcca
     4441 gaaacttgtg ctttctggat gtcctcctta aaggtgagag agtcatcatg gatcaactgc
     4501 atgaagttcc ggttggcaaa tccatcggac ttaagaaaat ccaggattgt ctttccactc
     4561 tgcttgtctc ggatcccatt gatcagtttt cttgacagcc gcccccatcc tgtatatcgg
     4621 cgcctcttga gctgtttcat gactttgtcg tcgaagagat gagcgtaagt tttcaagcgt
     4681 tcttcaatca tctccctatc ttcaaacaac gtaagggtga ggacaatgtc ctcaagaatg
     4741 tcctcgttct cctcattgtc caggaagtcc ttgtctttaa tgattttcag gagatcgtga
     4801 tacgttccca gggatgcgtt gaagcgatcc tccactccgc tgatttcaac agagtcgaaa
     4861 cattcaatct ttttgaaata gtcttctttg agctgtttca cggtaacttt ccggttcgtc
     4921 ttgaagagga ggtccacgat agctttcttc tgctctccag acaggaatgc tggctttctc
     4981 atcccttctg tgacgtattt gaccttggtg agctcgttat aaactgtgaa gtactcgtac
     5041 agcagagagt gtttaggaag caccttttcg ttaggcagat ttttatcaaa gttagtcatc
     5101 ctttcgatga aggactgggc agaggccccc ttatccacga cttcctcgaa gttccaggga
     5161 gtgatggtct cttctgattt gcgagtcatc cacgcgaatc tggaatttcc ccgggcgagg
     5221 gggcctacat agtagggtat ccgaaatgtg aggattttct caatcttttc cctgttatct
     5281 ttcaaaaagg ggtagaaatc ctcttgccgc ctgaggatag cgtgcagttc gcccaggtga
     5341 atctggtggg ggatgcttcc attgtcgaaa gtgcgctgtt tgcgcaacag atcttctctg
     5401 ttaagcttta ccagcagctc ctcggtgccg tccatttttt ccaagatggg cttaataaat
     5461 ttgtaaaatt cctcctggct tgctccgccg tcaatgtatc cggcgtagcc atttttagac
     5521 tgatcgaaga aaatttcctt gtacttctca ggcagttgct gtctgacaag ggccttcagc
     5581 aaagtcaagt cttggtggtg ctcatcatag cgcttgatca tactagcgct cagcggagct
     5641 ttggtgatct ccgtgttcac tcgcagaata tcactcagca gaatggcgtc tgacaggttc
     5701 tttgccgcca aaaaaaggtc tgcgtactgg tcgccgatct gggccagcag attgtcgaga
     5761 tcatcatcgt aggtgtcttt gctcagttga agcttggcat cttcggccag gtcgaagtta
     5821 gatttaaagt tgggggtcag cccgagtgac agggcgataa gattaccaaa caggccgttc
     5881 ttcttctccc cagggagctg tgcgatgagg ttttcgagcc gccgggattt ggacagccta
     5941 gcgctcagga ttgctttggc gtcaactccg gatgcgttga tcgggttctc ttcgaaaagc
     6001 tgattgtaag tctgaaccag ttggataaag agtttgtcga catcgctgtt gtctgggttc
     6061 aggtccccct cgatgaggaa gtgtccccga aatttgatca tatgcgccag cgcgagatag
     6121 atcaaccgca agtcagcctt atcagtactg tctacaagct tcttcctcag atgatatatg
     6181 gttgggtact tttcatggta cgccacctcg tccacgatat tgccaaagat tgggtggcgc
     6241 tcgtgctttt tatcctcctc caccaaaaag gactcctcca gcctatggaa gaaagagtca
     6301 tccaccttag ccatctcatt actaaagatc tcctgcaggt agcagatccg attctttctg
     6361 cgggtatatc tgcgccgtgc tgttcttttg agccgcgtgg cttcggccgt ctccccggag
     6421 tcgaacagga gggcgccaat gaggttcttc tttatgctgt ggcgatcggt attgcccaga
     6481 actttgaatt ttttgctcgg caccttgtac tcgtccgtaa tgacggccca gccgacgctg
     6541 tttgtgccga tagcgagccc aatggagtac ttcttgtcca tagagcctgc ttttttgtac
     6601 aaacttgttt gattcgaccc accctctttt tttgggtttg gtggggtatc ttcatcatcg
     6661 aatagatagt tatatacatc atccattgta gtggtattaa acatccctgt agtgattcca
     6721 aacgcgttat acgcagtttg gtccgtccaa ccaggtgaca gtggttttga attattacca
     6781 tcatcaattt tactagccgt gatttcatta ttcatgaagt tatcatgaac gttagaggag
     6841 gcaattggtt gtgaaagcgc ttgagaattt gtttgagttg ttatgaggtt cggaccgttg
     6901 ctactgttag tgaaagtgaa ggacaatgag ctatcagcaa tattcccact ttgattaaaa
     6961 ttggcgccgc tcgagacctt ccgcttcttc ttgggcatca ctagttctag aaaacttaga
     7021 ttagattgct atgctttctt tctaatgagc aagaagtaaa aaaagttgta atagaacaag
     7081 aaaaatgaaa ctgaaacttg agaaattgaa gaccgtttat taacttaaat atcaatggga
     7141 ggtcatcgaa agagaaaaaa atcaaaaaaa aaattttcaa gaaaaagaaa cgtgataaaa
     7201 atttttattg cctttttcga cgaagaaaaa gaaacgaggc ggtctctttt ttcttttcca
     7261 aacctttagt acgggtaatt aacgacaccc tagaggaaga aagaggggaa atttagtatg
     7321 ctgtgcttgg gtgttttgaa gtggtacggc gatgcgcgga gtccgagaaa atctggaaga
     7381 gtaaaaaagg agtagaaaca ttttgaagct atgagctcca gcttttgttc cctttagtga
     7441 gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgaa attgttatcc
     7501 gctcacaatt ccacacaaca taggagccgg aagcataaag tgtaaagcct ggggtgccta
     7561 atgagtgagg taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     7621 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     7681 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     7741 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     7801 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     7861 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     7921 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     7981 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     8041 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     8101 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     8161 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     8221 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     8281 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     8341 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     8401 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     8461 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     8521 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     8581 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     8641 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     8701 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     8761 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     8821 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     8881 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     8941 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     9001 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     9061 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     9121 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     9181 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     9241 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     9301 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     9361 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     9421 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     9481 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     9541 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     9601 tccccgaaaa gtgccacctg ggtccttttc atcacgtgct ataaaaataa ttataattta
     9661 aattttttaa tataaatata taaattaaaa atagaaagta aaaaaagaaa ttaaagaaaa
     9721 aatagttttt gttttccgaa gatgtaaaag actctagggg gatcgccaac aaatactacc
     9781 ttttatcttg ctcttcctgc tctcaggtat taatgccgaa ttgtttcatc ttgtctgtgt
     9841 agaagaccac acacgaaaat cctgtgattt tacattttac ttatcgttaa tcgaatgtat
     9901 atctatttaa tctgcttttc ttgtctaata aatatatatg taaagtacgc tttttgttga
     9961 aattttttaa acctttgttt attttttttt cttcattccg taactcttct accttcttta
    10021 tttactttct aaaatccaaa tacaaaacat aaaaataaat aaacacagag taaattccca
    10081 aattattcca tcattaaaag atacgaggcg cgtgtaagtt acaggcaagc gatccgtcct
    10141 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
    10201 gtc
//
