LOCUS       sequence-366133-       14407 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dCas9 fused with 5 copies of GCN4 epitope.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14407)
  AUTHORS   Chan WF, Coughlan HD, Chen Y, Keenan CR, Smyth GK, Perkins AC, 
            Johanson TM, Allan RS
  TITLE     Activation of stably silenced genes by recruitment of a synthetic 
            de-methylating module.
  JOURNAL   Nat Commun. 2022 Sep 23;13(1):5582. doi: 10.1038/s41467-022-33181-4.
  PUBMED    36151095
REFERENCE   2  (bases 1 to 14407)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2022-09-23"; volume: "13(1):5582. doi"; pages: " 
            10.1038/s41467-022-33181-4"
COMMENT     Sequence Label: dCas9-5xGCN4-P2A-BFP
FEATURES             Location/Qualifiers
     source          1..14407
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(182..200)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(183..200)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(490..509)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(723..742)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     842..864
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(902..920)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        988..1092
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1093..1161,1162..1953)
                     /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(1311..1330)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2124..2712
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2613..2632
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2866..2883
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2953..2973)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2958..3287
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3138..3273
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          4421..4486
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             4616..4636
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    5061..5195
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5098..5117)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5152..5171
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             5363..5996
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    6043..6168
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6665..6898
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7083..7127
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    7412..7529
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        7592..7802
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7661..7796
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             7818..7838
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      7887..7896
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(7893..7919,7920..7922,7923..10409,10410..10412,
                     10413..11996)
                     /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             7893..11996
                     /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             12000..12026
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             12006..12032
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     CDS             12045..12065
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             12072..12092
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             12156..12212
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             12279..12335
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             12402..12458
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             12525..12581
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             12648..12704
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             12795..12815
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             12825..12881
                     /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"
     CDS             12894..13592
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     CDS             13602..13622
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    13644..14232
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(13697..13717)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(14115..14126)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             join(14331..14407,1..157)
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
ORIGIN
        1 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
       61 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
      121 ctcagaccct tttagtcagt gtggaaaatc tctagcagca tctagaatta attccgtgta
      181 ttctatagtg tcacctaaat cgtatgtgta tgatacataa ggttatgtat taattgtagc
      241 cgcgttctaa cgacaatatg tacaagccta attgtgtagc atctggctta ctgaagcaga
      301 ccctatcatc tctctcgtaa actgccgtca gagtcggttt ggttggacga accttctgag
      361 tttctggtaa cgccgtcccg cacccggaaa tggtcagcga accaatcagc agggtcatcg
      421 ctagccagat cctctacgcc ggacgcatcg tggccggcat caccggcgcc acaggtgcgg
      481 ttgctggcgc ctatatcgcc gacatcaccg atggggaaga tcgggctcgc cacttcgggc
      541 tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg ggactgttgg
      601 gcgccatctc cttgcatgca ccattccttg cggcggcggt gctcaacggc ctcaacctac
      661 tactgggctg cttcctaatg caggagtcgc ataagggaga gcgtcgaatg gtgcactctc
      721 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
      781 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
      841 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
      901 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
      961 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     1021 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     1081 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     1141 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     1201 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     1261 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     1321 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     1381 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     1441 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     1501 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     1561 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     1621 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     1681 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     1741 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     1801 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     1861 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     1921 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     1981 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     2041 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     2101 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     2161 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     2221 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     2281 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     2341 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     2401 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     2461 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     2521 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     2581 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     2641 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     2701 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     2761 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     2821 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     2881 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     2941 taatgcagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3001 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3061 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3121 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3181 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     3241 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag cttggacaca
     3301 agacaggctt gcgagatatg tttgagaata ccactttatc ccgcgtcagg gagaggcagt
     3361 gcgtaaaaag acgcggactc atgtgaaata ctggttttta gtgcgccaga tctctataat
     3421 ctcgcgcaac ctattttccc ctcgaacact ttttaagccg tagataaaca ggctgggaca
     3481 cttcacatga gcgaaaaata catcgtcacc tgggacatgt tgcagatcca tgcacgtaaa
     3541 ctcgcaagcc gactgatgcc ttctgaacaa tggaaaggca ttattgccgt aagccgtggc
     3601 ggtctgtacc gggtgcgtta ctggcgcgtg aactgggtat tcgtcatgtc gataccgttt
     3661 gtatttccag ctacgatcac gacaaccagc gcgagcttaa agtgctgaaa cgcgcagaag
     3721 gcgatggcga aggcttcatc gttattgatg acctggtgga taccggtggt actgcggttg
     3781 cgattcgtga aatgtatcca aaagcgcact ttgtcaccat cttcgcaaaa ccggctggtc
     3841 gtccgctggt tgatgactat gttgttgata tcccgcaaga tacctggatt gaacagccgt
     3901 gggatatggg cgtcgtattc gtcccgccaa tctccggtcg ctaatctttt caacgcctgg
     3961 cactgccggg cgttgttctt tttaacttca ggcgggttac aatagtttcc agtaagtatt
     4021 ctggaggctg catccatgac acaggcaaac ctgagcgaaa ccctgttcaa accccgcttt
     4081 aaacatcctg aaacctcgac gctagtccgc cgctttaatc acggcgcaca accgcctgtg
     4141 cagtcggccc ttgatggtaa aaccatccct cactggtatc gcatgattaa ccgtctgatg
     4201 tggatctggc gcggcattga cccacgcgaa atcctcgacg tccaggcacg tattgtgatg
     4261 agcgatgccg aacgtaccga cgatgattta tacgatacgg tgattggcta ccgtggcggc
     4321 aactggattt atgagtgggc cccggatctt tgtgaaggaa ccttacttct gtggtgtgac
     4381 ataattggac aaactaccta cagagattta aagctctaag gtaaatataa aatttttaag
     4441 tgtataatgt gttaaactac tgattctaat tgtttgtgta ttttagattc caacctatgg
     4501 aactgatgaa tgggagcagt ggtggaatgc ctttaatgag gaaaacctgt tttgctcaga
     4561 agaaatgcca tctagtgatg atgaggctac tgctgactct caacattcta ctcctccaaa
     4621 aaagaagaga aaggtagaag accccaagga ctttccttca gaattgctaa gttttttgag
     4681 tcatgctgtg tttagtaata gaactcttgc ttgctttgct atttacacca caaaggaaaa
     4741 agctgcactg ctatacaaga aaattatgga aaaatattct gtaaccttta taagtaggca
     4801 taacagttat aatcataaca tactgttttt tcttactcca cacaggcata gagtgtctgc
     4861 tattaataac tatgctcaaa aattgtgtac ctttagcttt ttaatttgta aaggggttaa
     4921 taaggaatat ttgatgtata gtgccttgac tagagatcat aatcagccat accacatttg
     4981 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     5041 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     5101 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     5161 ccaaactcat caatgtatct tatcatgtct ggatcaactg gataactcaa gctaaccaaa
     5221 atcatcccaa acttcccacc ccatacccta ttaccactgc caattacctg tggtttcatt
     5281 tactctaaac ctgtgattcc tctgaattat tttcatttta aagaaattgt atttgttaaa
     5341 tatgtactac aaacttagta gttggaaggg ctaattcact cccaaagaag acaagatatc
     5401 cttgatctgt ggatctacca cacacaaggc tacttccctg attagcagaa ctacacacca
     5461 gggccagggg tcagatatcc actgaccttt ggatggtgct acaagctagt accagttgag
     5521 ccagataagg tagaagaggc caataaagga gagaacacca gcttgttaca ccctgtgagc
     5581 ctgcatggga tggatgaccc ggagagagaa gtgttagagt ggaggtttga cagccgccta
     5641 gcatttcatc acgtggcccg agagctgcat ccggagtact tcaagaactg ctgatatcga
     5701 gcttgctaca agggactttc cgctggggac tttccaggga ggcgtggcct gggcgggact
     5761 ggggagtggc gagccctcag atcctgcata taagcagctg ctttttgcct gtactgggtc
     5821 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
     5881 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
     5941 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg
     6001 cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact
     6061 cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa
     6121 attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg
     6181 gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat
     6241 aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc
     6301 ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag
     6361 acaggatcag aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat
     6421 caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac
     6481 aaaagtaaga ccaccgcaca gcaagcggcc ggccgctgat cttcagacct ggaggaggag
     6541 atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat
     6601 taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg
     6661 gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt
     6721 caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca
     6781 atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca
     6841 agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg
     6901 ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt
     6961 ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag
     7021 aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag
     7081 aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta
     7141 acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag
     7201 gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac
     7261 cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag
     7321 aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac
     7381 ggtatcgcca aatggcagta ttcatccaca attttaaaag aaaagggggg attggggggt
     7441 acagtgcagg ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac
     7501 aaaaacaaat tacaaaaatt caaaattttc gggtttatta cagggacagc agagatccag
     7561 tttggatcga taagcttgat atcgaattca ggcaggcaga agtatgcaaa gcatgcatct
     7621 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     7681 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     7741 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     7801 ctacgcgcgc caccatgccc aagaagaagc gcaaggtggg acgcgtctgc aggatatcaa
     7861 gcttgcggta ccgcgggccc gggatcgcca ccatggacaa gaagtacagc atcggcctgg
     7921 ccatcggcac caactctgtg ggctgggccg tgatcaccga cgagtacaag gtgcccagca
     7981 agaaattcaa ggtgctgggc aacaccgacc ggcacagcat caagaagaac ctgatcggcg
     8041 ccctgctgtt cgacagcgga gaaacagccg aggccacccg gctgaagaga accgccagaa
     8101 gaagatacac cagacggaag aaccggatct gctatctgca agagatcttc agcaacgaga
     8161 tggccaaggt ggacgacagc ttcttccaca gactggaaga gtccttcctg gtggaagagg
     8221 ataagaagca cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg
     8281 agaagtaccc caccatctac cacctgagaa agaaactggt ggacagcacc gacaaggccg
     8341 acctgcggct gatctatctg gccctggccc acatgatcaa gttccggggc cacttcctga
     8401 tcgagggcga cctgaacccc gacaacagcg acgtggacaa gctgttcatc cagctggtgc
     8461 agacctacaa ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg
     8521 ccatcctgtc tgccagactg agcaagagca gacggctgga aaatctgatc gcccagctgc
     8581 ccggcgagaa gaagaatggc ctgttcggca acctgattgc cctgagcctg ggcctgaccc
     8641 ccaacttcaa gagcaacttc gacctggccg aggatgccaa actgcagctg agcaaggaca
     8701 cctacgacga cgacctggac aacctgctgg cccagatcgg cgaccagtac gccgacctgt
     8761 ttctggccgc caagaacctg tccgacgcca tcctgctgag cgacatcctg agagtgaaca
     8821 ccgagatcac caaggccccc ctgagcgcct ctatgatcaa gagatacgac gagcaccacc
     8881 aggacctgac cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag tacaaagaga
     8941 ttttcttcga ccagagcaag aacggctacg ccggctacat cgatggcgga gccagccagg
     9001 aagagttcta caagttcatc aagcccatcc tggaaaagat ggacggcacc gaggaactgc
     9061 tcgtgaagct gaacagagag gacctgctgc ggaagcagcg gaccttcgac aacggcagca
     9121 tcccccacca gatccacctg ggagagctgc acgccattct gcggcggcag gaagattttt
     9181 acccattcct gaaggacaac cgggaaaaga tcgagaagat cctgaccttc cgcatcccct
     9241 actacgtggg ccctctggcc aggggaaaca gcagattcgc ctggatgacc agaaagagcg
     9301 aggaaaccat caccccctgg aacttcgagg aagtggtgga caagggcgcc agcgcccaga
     9361 gcttcatcga gcggatgacc aacttcgata agaacctgcc caacgagaag gtgctgccca
     9421 agcacagcct gctgtacgag tacttcaccg tgtacaacga gctgaccaaa gtgaaatacg
     9481 tgaccgaggg aatgagaaag cccgccttcc tgagcggcga gcagaaaaaa gccatcgtgg
     9541 acctgctgtt caagaccaac cggaaagtga ccgtgaagca gctgaaagag gactacttca
     9601 agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct
     9661 ccctgggcac ataccacgat ctgctgaaaa ttatcaagga caaggacttc ctggacaatg
     9721 aggaaaacga ggacattctg gaagatatcg tgctgaccct gacactgttt gaggacagag
     9781 agatgatcga ggaacggctg aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc
     9841 agctgaagcg gcggagatac accggctggg gcaggctgag ccggaagctg atcaacggca
     9901 tccgggacaa gcagtccggc aagacaatcc tggatttcct gaagtccgac ggcttcgcca
     9961 acagaaactt catgcagctg atccacgacg acagcctgac ctttaaagag gacatccaga
    10021 aagcccaggt gtccggccag ggcgatagcc tgcacgagca cattgccaat ctggccggca
    10081 gccccgccat taagaagggc atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag
    10141 tgatgggccg gcacaagccc gagaacatcg tgatcgaaat ggccagagag aaccagacca
    10201 cccagaaggg acagaagaac agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag
    10261 agctgggcag ccagatcctg aaagaacacc ccgtggaaaa cacccagctg cagaacgaga
    10321 agctgtacct gtactacctg cagaatgggc gggatatgta cgtggaccag gaactggaca
    10381 tcaaccggct gtccgactac gatgtggacg ctatcgtgcc tcagagcttt ctgaaggacg
    10441 actccatcga taacaaagtg ctgactcgga gcgacaagaa ccggggcaag agcgacaacg
    10501 tgccctccga agaggtcgtg aagaagatga agaactactg gcgccagctg ctgaatgcca
    10561 agctgattac ccagaggaag ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg
    10621 aactggataa ggccggcttc atcaagagac agctggtgga aacccggcag atcacaaagc
    10681 acgtggcaca gatcctggac tcccggatga acactaagta cgacgagaac gacaaactga
    10741 tccgggaagt gaaagtgatc accctgaagt ccaagctggt gtccgatttc cggaaggatt
    10801 tccagtttta caaagtgcgc gagatcaaca actaccacca cgcccacgac gcctacctga
    10861 acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt
    10921 acggcgacta caaggtgtac gacgtgcgga agatgatcgc caagagcgag caggaaatcg
    10981 gcaaggctac cgccaagtac ttcttctaca gcaacatcat gaactttttc aagaccgaga
    11041 ttaccctggc caacggcgag atccggaagc ggcctctgat cgagacaaac ggcgaaacag
    11101 gcgagatcgt gtgggataag ggccgggact ttgccaccgt gcggaaagtg ctgtctatgc
    11161 cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc aaagagtcta
    11221 tcctgcccaa gaggaacagc gacaagctga tcgccagaaa gaaggactgg gaccctaaga
    11281 agtacggcgg cttcgacagc cccaccgtgg cctattctgt gctggtggtg gccaaagtgg
    11341 aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct gctggggatc accatcatgg
    11401 aaagaagcag cttcgagaag aatcccatcg actttctgga agccaagggc tacaaagaag
    11461 tgaaaaagga cctgatcatc aagctgccta agtactccct gttcgagctg gaaaacggcc
    11521 ggaagagaat gctggcctct gccggcgaac tgcagaaggg aaacgaactg gccctgccct
    11581 ccaaatatgt gaacttcctg tacctggcca gccactatga gaagctgaag ggctcccccg
    11641 aggataatga gcagaaacag ctgtttgtgg aacagcacaa acactacctg gacgagatca
    11701 tcgagcagat cagcgagttc tccaagagag tgatcctggc cgacgctaat ctggacaagg
    11761 tgctgagcgc ctacaacaag cacagagaca agcctatcag agagcaggcc gagaatatca
    11821 tccacctgtt taccctgacc aatctgggag cccctgccgc cttcaagtac tttgacacca
    11881 ccatcgaccg gaagaggtac accagcacca aagaggtgct ggacgccacc ctgatccacc
    11941 agagcatcac cggcctgtac gagacacgga tcgacctgtc tcagctggga ggcgacgcct
    12001 atccctatga cgtgcccgat tatgccagcc tgggcagcgg ctcccccaag aaaaaacgca
    12061 aggtggaaga tcctaagaaa aagcggaaag tggacggcat tggtagtggg agcaacggca
    12121 gcagcggatc caacggtccg actgacgccg cggaagagga gcttctgagc aaaaactatc
    12181 acctcgaaaa cgaggttgcg cgactgaaga aaggaagcgg gtccggtgga agtggctccg
    12241 gatctggagg ttctggcagc ggaggtagcg gcagtggcga agagctcctt agtaagaact
    12301 atcatctgga aaatgaggta gcgcgcttaa agaaagggtc gggaagtggc ggcagcggaa
    12361 gtgggagtgg agggagcggt tctggcggtt ccggcagtgg agaggagttg ctgtctaaga
    12421 actaccactt agaaaacgaa gtcgcacggc taaaaaaagg ttccggctcc ggcggctccg
    12481 gttctggaag cgggggctcg ggatcaggtg gatctggatc aggagaggaa ttgctttcca
    12541 aaaactacca ccttgagaat gaggtggcca ggttaaagaa ggggagcggc tcggggggta
    12601 gtggatcggg gtcgggcggg tcaggaagcg gtggtagcgg atctggggag gagctgctct
    12661 cgaagaatta ccatttggag aacgaagtgg cgagactaaa gaagggaagc ggtagtggtg
    12721 gttcagggtc tggttcaggt ggcagtgggt ctgggggctc agggtccggg tcgcggccgc
    12781 aaggtggagg tggacccaag aagaagcgca aggtgggaag cggagctact aacttcagcc
    12841 tgctgaagca ggctggagac gtggaggaga accctggacc tgcaccagga agtatgagcg
    12901 agctgattaa ggagaacatg cacatgaagc tgtacatgga gggcaccgtg gacaaccatc
    12961 acttcaagtg cacatccgag ggcgaaggca agccctacga gggcacccag accatgagaa
    13021 tcaaggtggt cgagggcggc cctctcccct tcgccttcga catcctggct actagcttcc
    13081 tctacggcag caagaccttc atcaaccaca cccagggcat ccccgacttc ttcaagcagt
    13141 ccttccctga gggcttcaca tgggagagag tcaccacata cgaagacggg ggcgtgctga
    13201 ccgctaccca ggacaccagc ctccaggacg gctgcctcat ctacaacgtc aagatcagag
    13261 gggtgaactt cacatccaac ggccctgtga tgcagaagaa aacactcggc tgggaggcct
    13321 tcaccgagac gctgtacccc gctgacggcg gcctggaagg cagaaacgac atggccctga
    13381 agctcgtggg cgggagccat ctgatcgcaa acatcaagac cacatataga tccaagaaac
    13441 ccgctaagaa cctcaagatg cctggcgtct actatgtgga ctacagactg gaaagaatca
    13501 aggaggccaa caacgagacc tacgtcgagc agcacgaggt ggcagtggcc agatactgcg
    13561 acctccctag caaactgggg cacaagctta atggtggagg tccaaaaaag aaaaggaagg
    13621 tctaacctgc agggcttatc gataatcaac ctctggatta caaaatttgt gaaagattga
    13681 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
    13741 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
    13801 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
    13861 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
    13921 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
    13981 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
    14041 catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
    14101 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
    14161 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
    14221 ccgcctcccc gcatggtacc tgcagggatc gatcactgag gtacctttaa gaccaatgac
    14281 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
    14341 aattcactcc caacgaagac aagatctgct ttttgcttgt actgggtctc tctggttaga
    14401 ccagatc
//
