LOCUS       sequence-197550-       12163 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Full-length [FL] mouse Ezh2 fused to N-terminus of dCas9; pCDNA3 
            vector backbone, mammalian expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12163)
  AUTHORS   O'Geen H, Ren C, Nicolet CM, Perez AA, Halmai J, Le VM, Mackay JP, 
            Farnham PJ, Segal DJ
  TITLE     dCas9-based epigenome editing suggests acquisition of histone 
            methylation is not sufficient for target gene repression.
  JOURNAL   Nucleic Acids Res. 2017 Sep 29;45(17):9901-9916. doi: 
            10.1093/nar/gkx578.
  PUBMED    28973434
REFERENCE   2  (bases 1 to 12163)
  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"; doi: "10"; journalName: 
            "Nucleic Acids Res"; date: "2017-09-29- 29"; volume: "45"; issue: 
            "17"; pages: "9901-9916"
COMMENT     Sequence Label: Ezh2[FL]-dCas9
FEATURES             Location/Qualifiers
     source          1..12163
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             702..722
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             741..806
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             join(3135..3161,3162..3164,3165..5648,5649..5654,5655..5720,
                     5721..5723,5724..7238)
                     /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(3135..3161,3162..3164,3165..5651,5652..5654,
                     5655..7238)
                     /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             3135..7238
                     /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             7314..7361
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7380..7436
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     complement(7500..7519)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7544..7592
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7794..8222
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7881..7900)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8091..8112
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8231..8251)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8236..8565
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8416..8551
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8478..8497
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             8632..9426
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(8686..8705)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     9296..9315
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    9602..9723
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9639..9658)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9693..9712
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9772..9788)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9772..9788)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9785..9807)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9796..9812
                     /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(9820..9826,9827..9844,9845..9850))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9865..9886
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10003..10020)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10174..10762)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10254..10273)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10933..11724,11725..11793))
                     /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     11556..11575
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11794..11898)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(11973..11992)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
      661 ccagcctccg gactctagag gatcgaaccc ttgccaccat gcccaagaag aagaggaagg
      721 tgggaggctc cggaggaagc gactataagg accacgacgg agactacaag gatcatgata
      781 ttgattacaa agacgatgac gataagggtg gcgggtccgg cggtggatcc ggtaccatgg
      841 gccagactgg gaagaaatct gagaagggac cggtttgttg gcggaagcgt gtaaaatcag
      901 agtacatgag actgagacag ctcaagaggt tcagaagagc tgatgaagta aagactatgt
      961 ttagttccaa tcgtcagaaa attttggaaa gaactgaaac cttaaaccaa gagtggaagc
     1021 agcggaggat acagcctgtg cacatcatga cttctgtgag ctcattgcgc gggactaggg
     1081 agtgttcagt caccagtgac ttggattttc cagcacaagt catcccgtta aagaccctga
     1141 atgcagtcgc ctcggtgcct ataatgtact cttggtcgcc cttacaacag aattttatgg
     1201 tggaagacga aactgtttta cataacattc cttatatggg ggatgaagtt ctggatcagg
     1261 atggcacttt cattgaagaa ctaataaaaa attatgatgg aaaagtgcat ggtgacagag
     1321 aatgtggatt tataaatgat gaaatttttg tggagttggt aaatgctctt ggtcaatata
     1381 atgatgatga tgatgacgat gatggagatg atccagatga aagagaagaa aaacagaaag
     1441 atctagagga taatcgagat gataaagaaa cttgcccacc tcggaaattt cctgctgata
     1501 aaatatttga agccatttcc tcaatgtttc cagataaggg caccgcagaa gaactgaaag
     1561 aaaaatataa agaactcacg gagcagcagc tcccaggtgc tctgcctcct gaatgtactc
     1621 caaacatcga tggaccaaat gccaaatctg ttcagaggga gcaaagcttg cattcatttc
     1681 atacgctctt ctgtcgacga tgttttaagt atgactgctt cctacatccc ttccatgcaa
     1741 cacccaacac atataagagg aagaacacag aaacagcttt ggacaacaag ccttgtggac
     1801 cacagtgtta ccagcatctg gagggagcta aggagtttgc tgctgctctt actgctgagc
     1861 gtataaagac accacctaaa cgcccagggg gccgcagaag aggaagactt ccgaataaca
     1921 gtagcagacc cagcaccccc accatcagtg tgctggagtc aaaggataca gacagtgaca
     1981 gagaagcagg gactgaaact gggggagaga acaatgataa agaagaagaa gagaaaaaag
     2041 atgagacgtc cagctcctct gaagcaaatt ctcggtgtca aacaccaata aagatgaagc
     2101 caaatattga acctcctgag aatgtggagt ggagtggtgc tgaagcctcc atgtttagag
     2161 tcctcattgg tacttactac gataactttt gtgccattgc taggctaatt gggaccaaaa
     2221 catgtagaca ggtgtatgag tttagagtca aggagtccag tatcatagca cctgttccca
     2281 ctgaggatgt agacactcct ccaagaaaga agaaaaggaa acatcggttg tgggctgcac
     2341 actgcagaaa gatacaactg aaaaaggacg gctcctctaa ccatgtttac aactatcaac
     2401 cctgtgacca tccacggcag ccttgtgaca gttcgtgccc ttgtgtgata gcacaaaatt
     2461 tttgtgaaaa gttttgtcaa tgtagttcag agtgtcaaaa ccgctttcct ggatgtcggt
     2521 gcaaagcaca atgcaacacc aaacagtgtc catgctacct ggctgtccga gagtgtgacc
     2581 ctgacctctg tctcacgtgt ggagctgctg accattggga cagtaaaaat gtatcctgta
     2641 agaactgtag cattcagcgg ggctctaaaa agcacttact gctggcaccg tctgatgtgg
     2701 caggctgggg catctttatc aaagatcctg tacagaaaaa tgaattcatc tcagaatact
     2761 gtggggagat tatttctcag gatgaagcag acagaagagg aaaagtgtat gacaaataca
     2821 tgtgcagctt tctgttcaac ttgaacaatg attttgtggt ggatgcaacc cgaaagggca
     2881 acaaaattcg ttttgctaat cattcagtaa atccaaactg ctatgcaaaa gttatgatgg
     2941 ttaatggtga ccacaggata ggcatctttg ctaagagggc tatccagact ggtgaagagt
     3001 tgttttttga ttacagatac agccaggctg atgccctgaa gtatgtgggc atcgaacgag
     3061 aaatggaaat ccctagtacc ggagggagtg gtggaagcgg cggttctggt ggctcaggcg
     3121 gcagcggccg gccaatggac aagaagtact ccattgggct cgctatcggc acaaacagcg
     3181 tcggctgggc cgtcattacg gacgagtaca aggtgccgag caaaaaattc aaagttctgg
     3241 gcaataccga tcgccacagc ataaagaaga acctcattgg cgccctcctg ttcgactccg
     3301 gggagacggc cgaagccacg cggctcaaaa gaacagcacg gcgcagatat acccgcagaa
     3361 agaatcggat ctgctacctg caggagatct ttagtaatga gatggctaag gtggatgact
     3421 ctttcttcca taggctggag gagtcctttt tggtggagga ggataaaaag cacgagcgcc
     3481 acccaatctt tggcaatatc gtggacgagg tggcgtacca tgaaaagtac ccaaccatat
     3541 atcatctgag gaagaagctt gtagacagta ctgataaggc tgacttgcgg ttgatctatc
     3601 tcgcgctggc gcatatgatc aaatttcggg gacacttcct catcgagggg gacctgaacc
     3661 cagacaacag cgatgtcgac aaactcttta tccaactggt tcagacttac aatcagcttt
     3721 tcgaagagaa cccgatcaac gcatccggag ttgacgccaa agcaatcctg agcgctaggc
     3781 tgtccaaatc ccggcggctc gaaaacctca tcgcacagct ccctggggag aagaagaacg
     3841 gcctgtttgg taatcttatc gccctgtcac tcgggctgac ccccaacttt aaatctaact
     3901 tcgacctggc cgaagatgcc aagcttcaac tgagcaaaga cacctacgat gatgatctcg
     3961 acaatctgct ggcccagatc ggcgaccagt acgcagacct ttttttggcg gcaaagaacc
     4021 tgtcagacgc cattctgctg agtgatattc tgcgagtgaa cacggagatc accaaagctc
     4081 cgctgagcgc tagtatgatc aagcgctatg atgagcacca ccaagacttg actttgctga
     4141 aggcccttgt cagacagcaa ctgcctgaga agtacaagga aattttcttc gatcagtcta
     4201 aaaatggcta cgccggatac attgacggcg gagcaagcca ggaggaattt tacaaattta
     4261 ttaagcccat cttggaaaaa atggacggca ccgaggagct gctggtaaag cttaacagag
     4321 aagatctgtt gcgcaaacag cgcactttcg acaatggaag catcccccac cagattcacc
     4381 tgggcgaact gcacgctatc ctcaggcggc aagaggattt ctaccccttt ttgaaagata
     4441 acagggaaaa gattgagaaa atcctcacat ttcggatacc ctactatgta ggccccctcg
     4501 cccggggaaa ttccagattc gcgtggatga ctcgcaaatc agaagagacc atcactccct
     4561 ggaacttcga ggaagtcgtg gataaggggg cctctgccca gtccttcatc gaaaggatga
     4621 ctaactttga taaaaatctg cctaacgaaa aggtgcttcc taaacactct ctgctgtacg
     4681 agtacttcac agtttataac gagctcacca aggtcaaata cgtcacagaa gggatgagaa
     4741 agccagcatt cctgtctgga gagcagaaga aagctatcgt ggacctcctc ttcaagacga
     4801 accggaaagt taccgtgaaa cagctcaaag aagactattt caaaaagatt gaatgtttcg
     4861 actctgttga aatcagcgga gtggaggatc gcttcaacgc atccctggga acgtatcacg
     4921 atctcctgaa aatcattaaa gacaaggact tcctggacaa tgaggagaac gaggacattc
     4981 ttgaggacat tgtcctcacc cttacgttgt ttgaagatag ggagatgatt gaagaacgct
     5041 tgaaaactta cgctcatctc ttcgacgaca aagtcatgaa acagctcaag aggcgccgat
     5101 atacaggatg ggggcggctg tcaagaaaac tgatcaatgg gatccgagac aagcagagtg
     5161 gaaagacaat cctggatttt cttaagtccg atggatttgc caacaggaac ttcatgcagt
     5221 tgatccatga tgactctctc acctttaagg aggacatcca gaaagcacaa gtttctggcc
     5281 agggggacag tcttcacgag cacatcgcta atcttgcagg tagcccagct atcaaaaagg
     5341 gaatactgca gaccgttaag gtcgtggatg aactcgtcaa agtaatggga aggcataagc
     5401 ccgagaatat cgttatcgag atggcccgag agaaccaaac tacccagaag ggacagaaga
     5461 acagtaggga aaggatgaag aggattgaag agggtataaa agaactgggg tcccaaatcc
     5521 ttaaggaaca cccagttgaa aacacccagc ttcagaatga gaagctctac ctgtactacc
     5581 tgcagaacgg cagggacatg tacgtggatc aggaactgga catcaatcgg ctctccgact
     5641 acgacgtgga tgctatcgtg ccccagtctt ttctcaaaga tgattctatt gataataaag
     5701 tgttgacaag atccgataaa aatagaggga agagtgataa cgtcccctca gaagaagttg
     5761 tcaagaaaat gaaaaattat tggcggcagc tgctgaacgc caaactgatc acacaacgga
     5821 agttcgataa tctgactaag gctgaacgag gtggcctgtc tgagttggat aaagccggct
     5881 tcatcaaaag gcagcttgtt gagacacgcc agatcaccaa gcacgtggcc caaattctcg
     5941 attcacgcat gaacaccaag tacgatgaaa atgacaaact gattcgagag gtgaaagtta
     6001 ttactctgaa gtctaagctg gtctcagatt tcagaaagga ctttcagttt tataaggtga
     6061 gagagatcaa caattaccac catgcgcatg atgcctacct gaatgcagtg gtaggcactg
     6121 cacttatcaa aaaatatccc aagcttgaat ctgaatttgt ttacggagac tataaagtgt
     6181 acgatgttag gaaaatgatc gcaaagtctg agcaggaaat aggcaaggcc accgctaagt
     6241 acttctttta cagcaatatt atgaattttt tcaagaccga gattacactg gccaatggag
     6301 agattcggaa gcgaccactt atcgaaacaa acggagaaac aggagaaatc gtgtgggaca
     6361 agggtaggga tttcgcgaca gtccggaagg tcctgtccat gccgcaggtg aacatcgtta
     6421 aaaagaccga agtacagacc ggaggcttct ccaaggaaag tatcctcccg aaaaggaaca
     6481 gcgacaagct gatcgcacgc aaaaaagatt gggaccccaa gaaatacggc ggattcgatt
     6541 ctcctacagt cgcttacagt gtactggttg tggccaaagt ggagaaaggg aagtctaaaa
     6601 aactcaaaag cgtcaaggaa ctgctgggca tcacaatcat ggagcgatca agcttcgaaa
     6661 aaaaccccat cgactttctc gaggcgaaag gatataaaga ggtcaaaaaa gacctcatca
     6721 ttaagcttcc caagtactct ctctttgagc ttgaaaacgg ccggaaacga atgctcgcta
     6781 gtgcgggcga gctgcagaaa ggtaacgagc tggcactgcc ctctaaatac gttaatttct
     6841 tgtatctggc cagccactat gaaaagctca aagggtctcc cgaagataat gagcagaagc
     6901 agctgttcgt ggaacaacac aaacactacc ttgatgagat catcgagcaa ataagcgaat
     6961 tctccaaaag agtgatcctc gccgacgcta acctcgataa ggtgctttct gcttacaata
     7021 agcacaggga taagcccatc agggagcagg cagaaaacat tatccacttg tttactctga
     7081 ccaacttggg cgcgcctgca gccttcaagt acttcgacac caccatagac agaaagcggt
     7141 acacctctac aaaggaggtc ctggacgcca cactgattca tcagtcaatt acggggctct
     7201 atgaaacaag aatcgacctc tctcagctcg gtggagacgg tggtagtgga ggttcaggag
     7261 gatccggggg gagcggaggg agcgctagcg gtggaggctc cggaggtgga tctaaaaggc
     7321 cggcggccac gaaaaaggcc ggtcaggcaa aaaagaaaaa gggtggtagt ggaagcggag
     7381 ctactaactt cagcctgctg aagcaggctg gagacgtgga ggagaaccct ggacctgcgg
     7441 ccgcatgaaa gggttcgatc cctaccggtt agtaatgagt ttaaacgggg gaggctaact
     7501 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     7561 aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     7621 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     7681 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     7741 ggcaggccct gccatagcag atctgcgcag ctggggctct agggggtatc cccacgcgcc
     7801 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     7861 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     7921 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     7981 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     8041 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     8101 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     8161 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     8221 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     8281 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     8341 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     8401 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     8461 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     8521 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     8581 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     8641 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     8701 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     8761 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     8821 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     8881 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     8941 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     9001 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     9061 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     9121 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     9181 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     9241 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     9301 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     9361 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     9421 ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc
     9481 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     9541 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     9601 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     9661 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     9721 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     9781 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     9841 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     9901 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     9961 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
    10021 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
    10081 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
    10141 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
    10201 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
    10261 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
    10321 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
    10381 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
    10441 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
    10501 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
    10561 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
    10621 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
    10681 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
    10741 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
    10801 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
    10861 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
    10921 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
    10981 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
    11041 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
    11101 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
    11161 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
    11221 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
    11281 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
    11341 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
    11401 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
    11461 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
    11521 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
    11581 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
    11641 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
    11701 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
    11761 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
    11821 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
    11881 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
    11941 agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
    12001 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
    12061 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg
    12121 cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gtt
//
