LOCUS       sequence-462149-       11332 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  VPR activation domain coupled to catalytically inactive dCas9 for 
            localization; contains tdTomato for labeling and as a spacer.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11332)
  TITLE     HP1 binding creates a local barrier against transcription activation
            and persists during chromatin decondensation
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 11332)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pdCas9-tdTomato-VPR
FEATURES             Location/Qualifiers
     source          1..11332
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             635..655
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      704..713
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(710..736,737..739,740..3226,3227..3229,3230..4813)
                     /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             710..4813
                     /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             4817..4843
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4850..4870
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      4877..4886
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4883..6310
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     CDS             6344..6364
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6431..6580
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             6605..6625
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7055..7411
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALL"
     CDS             7430..7999
                     /codon_start=1
                     /product="transcriptional activation domain from the human 
                     herpesvirus 4 (Epstein-Barr virus) replication and 
                     transcription activator Rta/BRLF1 (Hardwick et al., 1992; 
                     Chavez et al., 2015)"
                     /label=Rta AD
                     /translation="RDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWAN
                     RPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKAL
                     REMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEIL
                     DTFLNDECLLHAMHISTGLSIFDTSLF"
     polyA_signal    8127..8248
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8164..8183)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8218..8237
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(8255..8710)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8392..8413)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8604..8623
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        8737..8841
                     /gene="bla"
                     /label=AmpR promoter
     promoter        8843..9200
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        8843..9034
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(8866..8886)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      9051..9186
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             9235..10029
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             9235..10029
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(9289..9308)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     9899..9918
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(10217..10236)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    10261..10308
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      10637..11225
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11126..11145
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgaca
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agcgctaccg gactcagatc tcgaggccac catgcccaag aagaagcgca aggtgggacg
      661 cgtctgcagg atatcaagct tgcggtaccg cgggcccggg atcgccacca tggacaagaa
      721 gtacagcatc ggcctggcca tcggcaccaa ctctgtgggc tgggccgtga tcaccgacga
      781 gtacaaggtg cccagcaaga aattcaaggt gctgggcaac accgaccggc acagcatcaa
      841 gaagaacctg atcggcgccc tgctgttcga cagcggagaa acagccgagg ccacccggct
      901 gaagagaacc gccagaagaa gatacaccag acggaagaac cggatctgct atctgcaaga
      961 gatcttcagc aacgagatgg ccaaggtgga cgacagcttc ttccacagac tggaagagtc
     1021 cttcctggtg gaagaggata agaagcacga gcggcacccc atcttcggca acatcgtgga
     1081 cgaggtggcc taccacgaga agtaccccac catctaccac ctgagaaaga aactggtgga
     1141 cagcaccgac aaggccgacc tgcggctgat ctatctggcc ctggcccaca tgatcaagtt
     1201 ccggggccac ttcctgatcg agggcgacct gaaccccgac aacagcgacg tggacaagct
     1261 gttcatccag ctggtgcaga cctacaacca gctgttcgag gaaaacccca tcaacgccag
     1321 cggcgtggac gccaaggcca tcctgtctgc cagactgagc aagagcagac ggctggaaaa
     1381 tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg ttcggcaacc tgattgccct
     1441 gagcctgggc ctgaccccca acttcaagag caacttcgac ctggccgagg atgccaaact
     1501 gcagctgagc aaggacacct acgacgacga cctggacaac ctgctggccc agatcggcga
     1561 ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc gacgccatcc tgctgagcga
     1621 catcctgaga gtgaacaccg agatcaccaa ggcccccctg agcgcctcta tgatcaagag
     1681 atacgacgag caccaccagg acctgaccct gctgaaagct ctcgtgcggc agcagctgcc
     1741 tgagaagtac aaagagattt tcttcgacca gagcaagaac ggctacgccg gctacatcga
     1801 tggcggagcc agccaggaag agttctacaa gttcatcaag cccatcctgg aaaagatgga
     1861 cggcaccgag gaactgctcg tgaagctgaa cagagaggac ctgctgcgga agcagcggac
     1921 cttcgacaac ggcagcatcc cccaccagat ccacctggga gagctgcacg ccattctgcg
     1981 gcggcaggaa gatttttacc cattcctgaa ggacaaccgg gaaaagatcg agaagatcct
     2041 gaccttccgc atcccctact acgtgggccc tctggccagg ggaaacagca gattcgcctg
     2101 gatgaccaga aagagcgagg aaaccatcac cccctggaac ttcgaggaag tggtggacaa
     2161 gggcgccagc gcccagagct tcatcgagcg gatgaccaac ttcgataaga acctgcccaa
     2221 cgagaaggtg ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt acaacgagct
     2281 gaccaaagtg aaatacgtga ccgagggaat gagaaagccc gccttcctga gcggcgagca
     2341 gaaaaaagcc atcgtggacc tgctgttcaa gaccaaccgg aaagtgaccg tgaagcagct
     2401 gaaagaggac tacttcaaga aaatcgagtg cttcgactcc gtggaaatct ccggcgtgga
     2461 agatcggttc aacgcctccc tgggcacata ccacgatctg ctgaaaatta tcaaggacaa
     2521 ggacttcctg gacaatgagg aaaacgagga cattctggaa gatatcgtgc tgaccctgac
     2581 actgtttgag gacagagaga tgatcgagga acggctgaaa acctatgccc acctgttcga
     2641 cgacaaagtg atgaagcagc tgaagcggcg gagatacacc ggctggggca ggctgagccg
     2701 gaagctgatc aacggcatcc gggacaagca gtccggcaag acaatcctgg atttcctgaa
     2761 gtccgacggc ttcgccaaca gaaacttcat gcagctgatc cacgacgaca gcctgacctt
     2821 taaagaggac atccagaaag cccaggtgtc cggccagggc gatagcctgc acgagcacat
     2881 tgccaatctg gccggcagcc ccgccattaa gaagggcatc ctgcagacag tgaaggtggt
     2941 ggacgagctc gtgaaagtga tgggccggca caagcccgag aacatcgtga tcgaaatggc
     3001 cagagagaac cagaccaccc agaagggaca gaagaacagc cgcgagagaa tgaagcggat
     3061 cgaagagggc atcaaagagc tgggcagcca gatcctgaaa gaacaccccg tggaaaacac
     3121 ccagctgcag aacgagaagc tgtacctgta ctacctgcag aatgggcggg atatgtacgt
     3181 ggaccaggaa ctggacatca accggctgtc cgactacgat gtggacgcta tcgtgcctca
     3241 gagctttctg aaggacgact ccatcgataa caaagtgctg actcggagcg acaagaaccg
     3301 gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag aagatgaaga actactggcg
     3361 ccagctgctg aatgccaagc tgattaccca gaggaagttc gacaatctga ccaaggccga
     3421 gagaggcggc ctgagcgaac tggataaggc cggcttcatc aagagacagc tggtggaaac
     3481 ccggcagatc acaaagcacg tggcacagat cctggactcc cggatgaaca ctaagtacga
     3541 cgagaacgac aaactgatcc gggaagtgaa agtgatcacc ctgaagtcca agctggtgtc
     3601 cgatttccgg aaggatttcc agttttacaa agtgcgcgag atcaacaact accaccacgc
     3661 ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg atcaaaaagt accctaagct
     3721 ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac gtgcggaaga tgatcgccaa
     3781 gagcgagcag gaaatcggca aggctaccgc caagtacttc ttctacagca acatcatgaa
     3841 ctttttcaag accgagatta ccctggccaa cggcgagatc cggaagcggc ctctgatcga
     3901 gacaaacggc gaaacaggcg agatcgtgtg ggataagggc cgggactttg ccaccgtgcg
     3961 gaaagtgctg tctatgcccc aagtgaatat cgtgaaaaag accgaggtgc agacaggcgg
     4021 cttcagcaaa gagtctatcc tgcccaagag gaacagcgac aagctgatcg ccagaaagaa
     4081 ggactgggac cctaagaagt acggcggctt cgacagcccc accgtggcct attctgtgct
     4141 ggtggtggcc aaagtggaaa agggcaagtc caagaaactg aagagtgtga aagagctgct
     4201 ggggatcacc atcatggaaa gaagcagctt cgagaagaat cccatcgact ttctggaagc
     4261 caagggctac aaagaagtga aaaaggacct gatcatcaag ctgcctaagt actccctgtt
     4321 cgagctggaa aacggccgga agagaatgct ggcctctgcc ggcgaactgc agaagggaaa
     4381 cgaactggcc ctgccctcca aatatgtgaa cttcctgtac ctggccagcc actatgagaa
     4441 gctgaagggc tcccccgagg ataatgagca gaaacagctg tttgtggaac agcacaaaca
     4501 ctacctggac gagatcatcg agcagatcag cgagttctcc aagagagtga tcctggccga
     4561 cgctaatctg gacaaggtgc tgagcgccta caacaagcac agagacaagc ctatcagaga
     4621 gcaggccgag aatatcatcc acctgtttac cctgaccaat ctgggagccc ctgccgcctt
     4681 caagtacttt gacaccacca tcgaccggaa gaggtacacc agcaccaaag aggtgctgga
     4741 cgccaccctg atccaccaga gcatcaccgg cctgtacgag acacggatcg acctgtctca
     4801 gctgggaggc gacgcctatc cctatgacgt gcccgattat gcaccggtcc caaaaaagaa
     4861 aagaaaagta ccggtcgcca ccatggtgag caagggcgag gaggtcatca aagagttcat
     4921 gcgcttcaag gtgcgcatgg agggctccat gaacggccac gagttcgaga tcgagggcga
     4981 gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggcgg
     5041 ccccctgccc ttcgcctggg acatcctgtc cccccagttc atgtacggct ccaaggcgta
     5101 cgtgaagcac cccgccgaca tccccgatta caagaagctg tccttccccg agggcttcaa
     5161 gtgggagcgc gtgatgaact tcgaggacgg cggtctggtg accgtgaccc aggactcctc
     5221 cctgcaggac ggcacgctga tctacaaggt gaagatgcgc ggcaccaact tcccccccga
     5281 cggccccgta atgcagaaga agaccatggg ctgggaggcc tccaccgagc gcctgtaccc
     5341 ccgcgacggc gtgctgaagg gcgagatcca ccaggccctg aagctgaagg acggcggcca
     5401 ctacctggtg gagttcaaga ccatctacat ggccaagaag cccgtgcaac tgcccggcta
     5461 ctactacgtg gacaccaagc tggacatcac ctcccacaac gaggactaca ccatcgtgga
     5521 acagtacgag cgctccgagg gccgccacca cctgttcctg gggcatggca ccggcagcac
     5581 cggcagcggc agctccggca ccgcctcctc cgaggacaac aacatggccg tcatcaaaga
     5641 gttcatgcgc ttcaaggtgc gcatggaggg ctccatgaac ggccacgagt tcgagatcga
     5701 gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa
     5761 gggcggcccc ctgcccttcg cctgggacat cctgtccccc cagttcatgt acggctccaa
     5821 ggcgtacgtg aagcaccccg ccgacatccc cgattacaag aagctgtcct tccccgaggg
     5881 cttcaagtgg gagcgcgtga tgaacttcga ggacggcggt ctggtgaccg tgacccagga
     5941 ctcctccctg caggacggca cgctgatcta caaggtgaag atgcgcggca ccaacttccc
     6001 ccccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcca ccgagcgcct
     6061 gtacccccgc gacggcgtgc tgaagggcga gatccaccag gccctgaagc tgaaggacgg
     6121 cggccactac ctggtggagt tcaagaccat ctacatggcc aagaagcccg tgcaactgcc
     6181 cggctactac tacgtggaca ccaagctgga catcacctcc cacaacgagg actacaccat
     6241 cgtggaacag tacgagcgct ccgagggccg ccaccacctg ttcctgtacg gcatggacga
     6301 gctgtacaag actagtcgca gggggcgtcg actgggtacc cctcctaaga agaagaggaa
     6361 ggttgccccc accatggata aaaagaagag aaaggccccc ccgaccgatg tcagcctggg
     6421 ggacgagctc gacgcattgg acgattttga tctggatatg ctgggaagtg acgccctcga
     6481 tgattttgac cttgacatgc ttggttcgga tgcccttgat gactttgacc tcgacatgct
     6541 cggcagtgac gcccttgatg atttcgacct ggacatgctg attaactcta gaagttccgg
     6601 atctccgaaa aagaaacgca aagttggtag ccagtacctg cccgacaccg acgaccggca
     6661 ccggatcgag gaaaagcgga agcggaccta cgagacattc aagagcatca tgaagaagtc
     6721 ccccttcagc ggccccaccg accctagacc tccacctaga agaatcgccg tgcccagcag
     6781 atccagcgcc agcgtgccaa aacctgcccc ccagccttac cccttcacca gcagcctgag
     6841 caccatcaac tacgacgagt tccctaccat ggtgttcccc agcggccaga tctctcaggc
     6901 ctctgctctg gctccagccc ctcctcaggt gctgcctcag gctcctgctc ctgcaccagc
     6961 tccagccatg gtgtctgcac tggctcaggc accagcaccc gtgcctgtgc tggctcctgg
     7021 acctccacag gctgtggctc caccagcccc taaacctaca caggccggcg agggcacact
     7081 gtctgaagct ctgctgcagc tgcagttcga cgacgaggat ctgggagccc tgctgggaaa
     7141 cagcaccgat cctgccgtgt tcaccgacct ggccagcgtg gacaacagcg agttccagca
     7201 gctgctgaac cagggcatcc ctgtggcccc tcacaccacc gagcccatgc tgatggaata
     7261 ccccgaggcc atcacccggc tcgtgacagg cgctcagagg cctcctgatc cagctcctgc
     7321 ccctctggga gcaccaggcc tgcctaatgg actgctgtct ggcgacgagg acttcagctc
     7381 tatcgccgat atggatttct cagccttgct gggctctggc agcggcagcc gggattccag
     7441 ggaagggatg tttttgccga agcctgaggc cggctccgct attagtgacg tgtttgaggg
     7501 ccgcgaggtg tgccagccaa aacgaatccg gccatttcat cctccaggaa gtccatgggc
     7561 caaccgccca ctccccgcca gcctcgcacc aacaccaacc ggtccagtac atgagccagt
     7621 cgggtcactg accccggcac cagtccctca gccactggat ccagcgcccg cagtgactcc
     7681 cgaggccagt cacctgttgg aggatcccga tgaagagacg agccaggctg tcaaagccct
     7741 tcgggagatg gccgatactg tgattcccca gaaggaagag gctgcaatct gtggccaaat
     7801 ggacctttcc catccgcccc caaggggcca tctggatgag ctgacaacca cacttgagtc
     7861 catgaccgag gatctgaacc tggactcacc cctgaccccg gaattgaacg agattctgga
     7921 taccttcctg aacgacgagt gcctcttgca tgccatgcat atcagcacag gactgtccat
     7981 cttcgacaca tctctgtttt aataagcggc cgcgactcta gatcataatc agccatacca
     8041 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     8101 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     8161 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     8221 gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta
     8281 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     8341 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     8401 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     8461 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     8521 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     8581 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     8641 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     8701 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     8761 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     8821 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     8881 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     8941 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     9001 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     9061 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     9121 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     9181 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     9241 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     9301 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     9361 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     9421 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     9481 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     9541 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     9601 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     9661 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     9721 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     9781 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     9841 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     9901 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     9961 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
    10021 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
    10081 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
    10141 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
    10201 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
    10261 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
    10321 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
    10381 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
    10441 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
    10501 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
    10561 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
    10621 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
    10681 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
    10741 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
    10801 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
    10861 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
    10921 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
    10981 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
    11041 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
    11101 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
    11161 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
    11221 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
    11281 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gc
//
