LOCUS       sequence-197562-       10864 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  DNMT3A 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 10864)
  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 10864)
  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: DNMT3A-dCas9
FEATURES             Location/Qualifiers
     source          1..10864
                     /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             843..1775
                     /codon_start=1
                     /product="catalytic domain of DNA methyltransferase 3A"
                     /label=DNMT3A catalytic domain
                     /translation="PSRLQMFFANNHDQEFDPPKVYPPVPAEKRKPIRVLSLFDGIATG
                     LLVLKDLGIQVDRYIASEVCEDSITVGMVRHQGKIMYVGDVRSVTQKHIQEWGPFDLVI
                     GGSPCNDLSIVNPARKGLYEGTGRLFFEFYRLLHDARPKEGDDRPFFWLFENVVAMGVS
                     DKRDISRFLESNPVMIDAKEVSAAHRARYFWGNLPGMNRPLASTVNDKLELQECLEHGR
                     IAKFSKVRTITTRSNSIKQGKDQHFPVFMNEKEDILWCTEMERVFGFPVHYTDVSNMSR
                     LARQRLLGRSWSVPVIRHLFAPLKEYFACV"
     CDS             join(1836..1862,1863..1865,1866..4349,4350..4355,4356..4421,
                     4422..4424,4425..5939)
                     /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(1836..1862,1863..1865,1866..4352,4353..4355,
                     4356..5939)
                     /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             1836..5939
                     /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             6015..6062
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6081..6137
                     /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(6201..6220)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6245..6293
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6495..6923
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6582..6601)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6792..6813
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6932..6952)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6937..7266
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7117..7252
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7179..7198
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             7333..8127
                     /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(7387..7406)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7997..8016
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    8303..8424
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8340..8359)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8394..8413
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8473..8489)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8473..8489)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8486..8508)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8497..8513
                     /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(8521..8527,8528..8545,8546..8551))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8566..8587
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8704..8721)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8875..9463)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8955..8974)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9634..10425,10426..10494))
                     /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     10257..10276
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10495..10599)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(10674..10693)
                     /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 ggtacccgcg
      841 ccccctcccg gctccagatg ttcttcgcta ataaccacga ccaggaattt gaccctccaa
      901 aggtttaccc acctgtccca gctgagaaga ggaagcccat ccgggtgctg tctctctttg
      961 atggaatcgc tacagggctc ctggtgctga aggacttggg cattcaggtg gaccgctaca
     1021 ttgcctcgga ggtgtgtgag gactccatca cggtgggcat ggtgcggcac caggggaaga
     1081 tcatgtacgt cggggacgtc cgcagcgtca cacagaagca tatccaggag tggggcccat
     1141 tcgatctggt gattgggggc agtccctgca atgacctctc catcgtcaac cctgctcgca
     1201 agggcctcta cgagggcact ggccggctct tctttgagtt ctaccgcctc ctgcatgatg
     1261 cgcggcccaa ggagggagat gatcgcccct tcttctggct ctttgagaat gtggtggcca
     1321 tgggcgttag tgacaagagg gacatctcgc gatttctcga gtccaaccct gtgatgattg
     1381 atgccaaaga agtgtcagct gcacacaggg cccgctactt ctggggtaac cttcccggta
     1441 tgaacaggcc gttggcatcc actgtgaatg ataagctgga gctgcaggag tgtctggagc
     1501 atggcaggat agccaagttc agcaaagtga ggaccattac tacgaggtca aactccataa
     1561 agcagggcaa agaccagcat tttcctgtct tcatgaatga gaaagaggac atcttatggt
     1621 gcactgaaat ggaaagggta tttggtttcc cagtccacta tactgacgtc tccaacatga
     1681 gccgcttggc gaggcagaga ctgctgggcc ggtcatggag cgtgccagtc atccgccacc
     1741 tcttcgctcc gctgaaggag tattttgcgt gtgtgagtac cggagggagt ggtggaagcg
     1801 gcggttctgg tggctcaggc ggcagcggcc ggccaatgga caagaagtac tccattgggc
     1861 tcgctatcgg cacaaacagc gtcggctggg ccgtcattac ggacgagtac aaggtgccga
     1921 gcaaaaaatt caaagttctg ggcaataccg atcgccacag cataaagaag aacctcattg
     1981 gcgccctcct gttcgactcc ggggagacgg ccgaagccac gcggctcaaa agaacagcac
     2041 ggcgcagata tacccgcaga aagaatcgga tctgctacct gcaggagatc tttagtaatg
     2101 agatggctaa ggtggatgac tctttcttcc ataggctgga ggagtccttt ttggtggagg
     2161 aggataaaaa gcacgagcgc cacccaatct ttggcaatat cgtggacgag gtggcgtacc
     2221 atgaaaagta cccaaccata tatcatctga ggaagaagct tgtagacagt actgataagg
     2281 ctgacttgcg gttgatctat ctcgcgctgg cgcatatgat caaatttcgg ggacacttcc
     2341 tcatcgaggg ggacctgaac ccagacaaca gcgatgtcga caaactcttt atccaactgg
     2401 ttcagactta caatcagctt ttcgaagaga acccgatcaa cgcatccgga gttgacgcca
     2461 aagcaatcct gagcgctagg ctgtccaaat cccggcggct cgaaaacctc atcgcacagc
     2521 tccctgggga gaagaagaac ggcctgtttg gtaatcttat cgccctgtca ctcgggctga
     2581 cccccaactt taaatctaac ttcgacctgg ccgaagatgc caagcttcaa ctgagcaaag
     2641 acacctacga tgatgatctc gacaatctgc tggcccagat cggcgaccag tacgcagacc
     2701 tttttttggc ggcaaagaac ctgtcagacg ccattctgct gagtgatatt ctgcgagtga
     2761 acacggagat caccaaagct ccgctgagcg ctagtatgat caagcgctat gatgagcacc
     2821 accaagactt gactttgctg aaggcccttg tcagacagca actgcctgag aagtacaagg
     2881 aaattttctt cgatcagtct aaaaatggct acgccggata cattgacggc ggagcaagcc
     2941 aggaggaatt ttacaaattt attaagccca tcttggaaaa aatggacggc accgaggagc
     3001 tgctggtaaa gcttaacaga gaagatctgt tgcgcaaaca gcgcactttc gacaatggaa
     3061 gcatccccca ccagattcac ctgggcgaac tgcacgctat cctcaggcgg caagaggatt
     3121 tctacccctt tttgaaagat aacagggaaa agattgagaa aatcctcaca tttcggatac
     3181 cctactatgt aggccccctc gcccggggaa attccagatt cgcgtggatg actcgcaaat
     3241 cagaagagac catcactccc tggaacttcg aggaagtcgt ggataagggg gcctctgccc
     3301 agtccttcat cgaaaggatg actaactttg ataaaaatct gcctaacgaa aaggtgcttc
     3361 ctaaacactc tctgctgtac gagtacttca cagtttataa cgagctcacc aaggtcaaat
     3421 acgtcacaga agggatgaga aagccagcat tcctgtctgg agagcagaag aaagctatcg
     3481 tggacctcct cttcaagacg aaccggaaag ttaccgtgaa acagctcaaa gaagactatt
     3541 tcaaaaagat tgaatgtttc gactctgttg aaatcagcgg agtggaggat cgcttcaacg
     3601 catccctggg aacgtatcac gatctcctga aaatcattaa agacaaggac ttcctggaca
     3661 atgaggagaa cgaggacatt cttgaggaca ttgtcctcac ccttacgttg tttgaagata
     3721 gggagatgat tgaagaacgc ttgaaaactt acgctcatct cttcgacgac aaagtcatga
     3781 aacagctcaa gaggcgccga tatacaggat gggggcggct gtcaagaaaa ctgatcaatg
     3841 ggatccgaga caagcagagt ggaaagacaa tcctggattt tcttaagtcc gatggatttg
     3901 ccaacaggaa cttcatgcag ttgatccatg atgactctct cacctttaag gaggacatcc
     3961 agaaagcaca agtttctggc cagggggaca gtcttcacga gcacatcgct aatcttgcag
     4021 gtagcccagc tatcaaaaag ggaatactgc agaccgttaa ggtcgtggat gaactcgtca
     4081 aagtaatggg aaggcataag cccgagaata tcgttatcga gatggcccga gagaaccaaa
     4141 ctacccagaa gggacagaag aacagtaggg aaaggatgaa gaggattgaa gagggtataa
     4201 aagaactggg gtcccaaatc cttaaggaac acccagttga aaacacccag cttcagaatg
     4261 agaagctcta cctgtactac ctgcagaacg gcagggacat gtacgtggat caggaactgg
     4321 acatcaatcg gctctccgac tacgacgtgg atgctatcgt gccccagtct tttctcaaag
     4381 atgattctat tgataataaa gtgttgacaa gatccgataa aaatagaggg aagagtgata
     4441 acgtcccctc agaagaagtt gtcaagaaaa tgaaaaatta ttggcggcag ctgctgaacg
     4501 ccaaactgat cacacaacgg aagttcgata atctgactaa ggctgaacga ggtggcctgt
     4561 ctgagttgga taaagccggc ttcatcaaaa ggcagcttgt tgagacacgc cagatcacca
     4621 agcacgtggc ccaaattctc gattcacgca tgaacaccaa gtacgatgaa aatgacaaac
     4681 tgattcgaga ggtgaaagtt attactctga agtctaagct ggtctcagat ttcagaaagg
     4741 actttcagtt ttataaggtg agagagatca acaattacca ccatgcgcat gatgcctacc
     4801 tgaatgcagt ggtaggcact gcacttatca aaaaatatcc caagcttgaa tctgaatttg
     4861 tttacggaga ctataaagtg tacgatgtta ggaaaatgat cgcaaagtct gagcaggaaa
     4921 taggcaaggc caccgctaag tacttctttt acagcaatat tatgaatttt ttcaagaccg
     4981 agattacact ggccaatgga gagattcgga agcgaccact tatcgaaaca aacggagaaa
     5041 caggagaaat cgtgtgggac aagggtaggg atttcgcgac agtccggaag gtcctgtcca
     5101 tgccgcaggt gaacatcgtt aaaaagaccg aagtacagac cggaggcttc tccaaggaaa
     5161 gtatcctccc gaaaaggaac agcgacaagc tgatcgcacg caaaaaagat tgggacccca
     5221 agaaatacgg cggattcgat tctcctacag tcgcttacag tgtactggtt gtggccaaag
     5281 tggagaaagg gaagtctaaa aaactcaaaa gcgtcaagga actgctgggc atcacaatca
     5341 tggagcgatc aagcttcgaa aaaaacccca tcgactttct cgaggcgaaa ggatataaag
     5401 aggtcaaaaa agacctcatc attaagcttc ccaagtactc tctctttgag cttgaaaacg
     5461 gccggaaacg aatgctcgct agtgcgggcg agctgcagaa aggtaacgag ctggcactgc
     5521 cctctaaata cgttaatttc ttgtatctgg ccagccacta tgaaaagctc aaagggtctc
     5581 ccgaagataa tgagcagaag cagctgttcg tggaacaaca caaacactac cttgatgaga
     5641 tcatcgagca aataagcgaa ttctccaaaa gagtgatcct cgccgacgct aacctcgata
     5701 aggtgctttc tgcttacaat aagcacaggg ataagcccat cagggagcag gcagaaaaca
     5761 ttatccactt gtttactctg accaacttgg gcgcgcctgc agccttcaag tacttcgaca
     5821 ccaccataga cagaaagcgg tacacctcta caaaggaggt cctggacgcc acactgattc
     5881 atcagtcaat tacggggctc tatgaaacaa gaatcgacct ctctcagctc ggtggagacg
     5941 gtggtagtgg aggttcagga ggatccgggg ggagcggagg gagcgctagc ggtggaggct
     6001 ccggaggtgg atctaaaagg ccggcggcca cgaaaaaggc cggtcaggca aaaaagaaaa
     6061 agggtggtag tggaagcgga gctactaact tcagcctgct gaagcaggct ggagacgtgg
     6121 aggagaaccc tggacctgcg gccgcatgaa agggttcgat ccctaccggt tagtaatgag
     6181 tttaaacggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     6241 atgacggcaa taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg
     6301 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     6361 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     6421 gctcgcagcc aacgtcgggg cggcaggccc tgccatagca gatctgcgca gctggggctc
     6481 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     6541 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     6601 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     6661 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     6721 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     6781 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     6841 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     6901 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     6961 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     7021 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     7081 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     7141 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     7201 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     7261 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     7321 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     7381 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     7441 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     7501 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     7561 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     7621 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     7681 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     7741 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     7801 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     7861 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     7921 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     7981 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     8041 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     8101 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa tgaccgacca
     8161 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     8221 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     8281 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     8341 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     8401 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     8461 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     8521 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     8581 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     8641 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     8701 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     8761 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     8821 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     8881 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     8941 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     9001 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     9061 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     9121 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     9181 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     9241 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     9301 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     9361 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     9421 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     9481 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     9541 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     9601 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     9661 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     9721 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     9781 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     9841 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     9901 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     9961 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
    10021 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
    10081 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
    10141 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
    10201 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
    10261 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
    10321 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
    10381 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
    10441 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
    10501 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
    10561 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
    10621 acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc
    10681 tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct
    10741 gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg
    10801 aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg
    10861 cgtt
//
