LOCUS       sequence-174076-        6204 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Targeted CpG methylation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6204)
  AUTHORS   Lei Y, Zhang X, Su J, Jeong M, Gundry MC, Huang YH, Zhou Y, Li W, 
            Goodell MA
  TITLE     Targeted DNA methylation in vivo using an engineered dCas9-MQ1 
            fusion protein.
  JOURNAL   Nat Commun. 2017 Jul 11;8:16026. doi: 10.1038/ncomms16026.
  PUBMED    28695892
REFERENCE   2  (bases 1 to 6204)
  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: "2017-07-11"; volume: "8:16026. doi"; pages: " 
            10.1038/ncomms16026"
COMMENT     Sequence Label: pLV hUbC-dCas9-dMQ1-EGFG
FEATURES             Location/Qualifiers
     source          1..6204
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      7..16
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             16..81
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             88..108
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(118..144,145..147,148..2631,2632..2637,2638..2703,
                     2704..2706,2707..4221)
                     /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(118..144,145..147,148..2634,2635..2637,2638..4221)
                     /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             118..4221
                     /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             4234..4254
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5425..5478
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(5485..5487,5488..6198)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(5485..5487,5488..6198)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(5527..5548)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5788..5807)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6135..6156
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 tctagagcca ccatggacta caaagaccat gacggtgatt ataaagatca tgacatcgat
       61 tacaaggatg acgatgacaa gatggccccc aagaagaaga ggaaggtggg ccgcggaatg
      121 gacaagaagt actccattgg gctcgccatc ggcacaaaca gcgtcggctg ggccgtcatt
      181 acggacgagt acaaggtgcc gagcaaaaaa ttcaaagttc tgggcaatac cgatcgccac
      241 agcataaaga agaacctcat tggcgccctc ctgttcgact ccggggaaac cgccgaagcc
      301 acgcggctca aaagaacagc acggcgcaga tatacccgca gaaagaatcg gatctgctac
      361 ctgcaggaga tctttagtaa tgagatggct aaggtggatg actctttctt ccataggctg
      421 gaggagtcct ttttggtgga ggaggataaa aagcacgagc gccacccaat ctttggcaat
      481 atcgtggacg aggtggcgta ccatgaaaag tacccaacca tatatcatct gaggaagaag
      541 cttgtagaca gtactgataa ggctgacttg cggttgatct atctcgcgct ggcgcatatg
      601 atcaaatttc ggggacactt cctcatcgag ggggacctga acccagacaa cagcgatgtc
      661 gacaaactct ttatccaact ggttcagact tacaatcagc ttttcgaaga gaacccgatc
      721 aacgcatccg gagttgacgc caaagcaatc ctgagcgcta ggctgtccaa atcccggcgg
      781 ctcgaaaacc tcatcgcaca gctccctggg gagaagaaga acggcctgtt tggtaatctt
      841 atcgccctgt cactcgggct gacccccaac tttaaatcta acttcgacct ggccgaagat
      901 gccaagcttc aactgagcaa agacacctac gatgatgatc tcgacaatct gctggcccag
      961 atcggcgacc agtacgcaga cctttttttg gcggcaaaga acctgtcaga cgccattctg
     1021 ctgagtgata ttctgcgagt gaacacggag atcaccaaag ctccgctgag cgctagtatg
     1081 atcaagcgct atgatgagca ccaccaagac ttgactttgc tgaaggccct tgtcagacag
     1141 caactgcctg agaagtacaa ggaaattttc ttcgatcagt ctaaaaatgg ctacgccgga
     1201 tacattgacg gcggagcaag ccaggaggaa ttttacaaat ttattaagcc catcttggaa
     1261 aaaatggacg gcaccgagga gctgctggta aagcttaaca gagaagatct gttgcgcaaa
     1321 cagcgcactt tcgacaatgg aagcatcccc caccagattc acctgggcga actgcacgct
     1381 atcctcaggc ggcaagagga tttctacccc tttttgaaag ataacaggga aaagattgag
     1441 aaaatcctca catttcggat accctactat gtaggccccc tcgcccgggg aaattccaga
     1501 ttcgcgtgga tgactcgcaa atcagaagag accatcactc cctggaactt cgaggaagtc
     1561 gtggataagg gggcctctgc ccagtccttc atcgaaagga tgactaactt tgataaaaat
     1621 ctgcctaacg aaaaggtgct tcctaaacac tctctgctgt acgagtactt cacagtttat
     1681 aacgagctca ccaaggtcaa atacgtcaca gaagggatga gaaagccagc attcctgtct
     1741 ggagagcaga agaaagctat cgtggacctc ctcttcaaga cgaaccggaa agttaccgtg
     1801 aaacagctca aagaagacta tttcaaaaag attgaatgtt tcgactctgt tgaaatcagc
     1861 ggagtggagg atcgcttcaa cgcatccctg ggaacgtatc acgatctcct gaaaatcatt
     1921 aaagacaagg acttcctgga caatgaggag aacgaggaca ttcttgagga cattgtcctc
     1981 acccttacgt tgtttgaaga tagggagatg attgaagaac gcttgaaaac ttacgctcat
     2041 ctcttcgacg acaaagtcat gaaacagctc aagaggcgcc gatatacagg atgggggcgg
     2101 ctgtcaagaa aactgatcaa tgggatccga gacaagcaga gtggaaagac aatcctggat
     2161 tttcttaagt ccgatggatt tgccaaccgg aacttcatgc agttgatcca tgatgactct
     2221 ctcaccttta aggaggacat ccagaaagca caagtttctg gccaggggga cagtcttcac
     2281 gagcacatcg ctaatcttgc aggtagccca gctatcaaaa agggaatact gcagaccgtt
     2341 aaggtcgtgg atgaactcgt caaagtaatg ggaaggcata agcccgagaa tatcgttatc
     2401 gagatggccc gagagaacca aactacccag aagggacaga agaacagtag ggaaaggatg
     2461 aagaggattg aagagggtat aaaagaactg gggtcccaaa tccttaagga acacccagtt
     2521 gaaaacaccc agcttcagaa tgagaagctc tacctgtact acctgcagaa cggcagggac
     2581 atgtacgtgg atcaggaact ggacatcaat cggctctccg actacgacgt ggatgccatc
     2641 gtgccccagt cttttctcaa agatgattct attgataata aagtgttgac aagatccgat
     2701 aaaaatagag ggaagagtga taacgtcccc tcagaagaag ttgtcaagaa aatgaaaaat
     2761 tattggcggc agctgctgaa cgccaaactg atcacacaac ggaagttcga taatctgact
     2821 aaggctgaac gaggtggcct gtctgagttg gataaagccg gcttcatcaa aaggcagctt
     2881 gttgagacac gccagatcac caagcacgtg gcccaaattc tcgattcacg catgaacacc
     2941 aagtacgatg aaaatgacaa actgattcga gaggtgaaag ttattactct gaagtctaag
     3001 ctggtctcag atttcagaaa ggactttcag ttttataagg tgagagagat caacaattac
     3061 caccatgcgc atgatgccta cctgaatgca gtggtaggca ctgcacttat caaaaaatat
     3121 cccaagcttg aatctgaatt tgtttacgga gactataaag tgtacgatgt taggaaaatg
     3181 atcgcaaagt ctgagcagga aataggcaag gccaccgcta agtacttctt ttacagcaat
     3241 attatgaatt ttttcaagac cgagattaca ctggccaatg gagagattcg gaagcgacca
     3301 cttatcgaaa caaacggaga aacaggagaa atcgtgtggg acaagggtag ggatttcgcg
     3361 acagtccgga aggtcctgtc catgccgcag gtgaacatcg ttaaaaagac cgaagtacag
     3421 accggaggct tctccaagga aagtatcctc ccgaaaagga acagcgacaa gctgatcgca
     3481 cgcaaaaaag attgggaccc caagaaatac ggcggattcg attctcctac agtcgcttac
     3541 agtgtactgg ttgtggccaa agtggagaaa gggaagtcta aaaaactcaa aagcgtcaag
     3601 gaactgctgg gcatcacaat catggagcga tcaagcttcg aaaaaaaccc catcgacttt
     3661 ctcgaggcga aaggatataa agaggtcaaa aaagacctca tcattaagct tcccaagtac
     3721 tctctctttg agcttgaaaa cggccggaaa cgaatgctcg ctagtgcggg cgagctgcag
     3781 aaaggtaacg agctggcact gccctctaaa tacgttaatt tcttgtatct ggccagccac
     3841 tatgaaaagc tcaaagggtc tcccgaagat aatgagcaga agcagctgtt cgtggaacaa
     3901 cacaaacact accttgatga gatcatcgag caaataagcg aattctccaa aagagtgatc
     3961 ctcgccgacg ctaacctcga taaggtgctt tctgcttaca ataagcacag ggataagccc
     4021 atcagggagc aggcagaaaa cattatccac ttgtttactc tgaccaactt gggcgcgcct
     4081 gcagccttca agtacttcga caccaccata gacagaaagc ggtacacctc tacaaaggag
     4141 gtcctggacg ccacactgat tcatcagtca attacggggc tctatgaaac aagaatcgac
     4201 ctctctcagc tcggtggaga cagcagggct gaccccaaga agaagaggaa ggtggctagg
     4261 gacagcaaag tggagaacaa aacaaagaag ctgagagtgt tcgaagcctt cgccggcatt
     4321 ggcgcccaga gaaaggccct ggagaaagtg aggaaggacg agtacgagat cgtgggactg
     4381 gccgagtggt atgtgcccgc catcgtcatg taccaggcca tccataacaa cttccacacc
     4441 aagctcgagt acaagtccgt cagcagagag gagatgatcg actacctcga gaacaagacc
     4501 ctgtcctgga acagcaagaa ccccgtcagc aacggatact ggaagaggaa gaaggatgac
     4561 gagctgaaga tcatctacaa cgccatcaag ctgtccgaaa aggagggcaa cattttcgac
     4621 atcagggacc tctacaagag gacactgaag aacatcgacc tgctcaccta cagcttccct
     4681 tcccaggacc tgagccagca aggcatccag aagggcatga agaggggaag cggcaccaga
     4741 tccggcctgc tctgggagat cgaaagagcc ctggactcca ccgagaagaa cgacctgcct
     4801 aagtacctcc tcatggagaa cgtgggagcc ctgctgcaca agaagaacga ggaggagctg
     4861 aaccaatgga agcagaagct ggagagcctg ggctaccaga acagcatcga agtcctcaat
     4921 gctgccgatt tcggatccag ccaggccagg aggagagtgt tcatgatctc caccctcaat
     4981 gagttcgtgg aactgcctaa gggcgacaag aagcccaaga gcatcaaaaa ggtgctgaac
     5041 aagatcgtga gcgagaagga catcctcaac aacctgctga aatacaacct caccgaattc
     5101 aagaagacca agtccaacat caacaaggcc agcctgatcg gctactccaa gttcaactcc
     5161 gagggctatg tgtacgaccc cgagttcaca ggccccacac tgacagctgc cggcgccaac
     5221 tccaggatca agatcaagga cggcagcaac atcaggaaga tgaacagcga cgagaccttc
     5281 ctgtacatcg gctttgacag ccaggacggc aagagagtga acgaaatcga gttcctgacc
     5341 gagaaccaga agatcttcgt gtgtggcaac agcatcagcg tggaggtgct ggaggccatc
     5401 attgacaaga tcggcggccc tagcgagggc agaggaagtc ttctaacatg cggtgacgtg
     5461 gaggagaatc ccggccctgg taccgtgagc aagggcgagg agctgttcac cggggtggtg
     5521 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     5581 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     5641 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     5701 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     5761 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     5821 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     5881 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     5941 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     6001 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     6061 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     6121 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     6181 atggacgagc tgtacaagac cggt
//
