LOCUS       sequence-79086-e        9689 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  encodes human-optimized dCas9_VP64 synthetic transcription factor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9689)
  AUTHORS   Farzadfard F, Perli SD, Lu TK
  TITLE     Tunable and Multifunctional Eukaryotic Transcription Factors Based 
            on CRISPR/Cas.
  JOURNAL   ACS Synth Biol. 2013 Sep 11.
  PUBMED    23977949
REFERENCE   2  (bases 1 to 9689)
  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: "ACS Synth 
            Biol. 2013 Sep 11."
COMMENT     Sequence Label: pCMV_dCas9_VP64
FEATURES             Location/Qualifiers
     source          1..9689
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(106..125)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    150..197
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     complement(518..535)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(689..1277)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(769..788)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1448..2239,2240..2308))
                     /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     2071..2090
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2309..2413)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2440..2895
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2527..2546)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2737..2758
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     enhancer        3084..3387
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3388..3591
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3542..3562
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3620..3629
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3626..3652,3653..3655,3656..6139,6140..6145,6146..6211,
                     6212..6214,6215..7729)
                     /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(3626..3652,3653..3655,3656..6142,6143..6145,
                     6146..7729)
                     /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             3626..7729
                     /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             7766..7786
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7793..7813
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7820..7840
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7880..8029
                     /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"
     polyA_signal    8200..8321
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8211..8230)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     8265..8284
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        8433..8932
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(8468..8491)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     8865..8884
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      8950..8959
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             8956..9669
                     /codon_start=1
                     /product="monomeric derivative of red fluorescent protein 
                     from Entacmaea quadricolor (Merzlyak et al., 2007)"
                     /label=TagRFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTEMLYPADGGLEGRSD
                     MALKLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
ORIGIN
        1 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
       61 ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga
      121 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
      181 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
      241 cccaccgaga cgccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
      301 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
      361 agcctcagct cgagtatgtc gaccgatgcc cttgagagcc ttcaacccag tcagctcctt
      421 ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct ttatcatgca
      481 actcgtagga caggtgccgg cagcgctctt ccgcttcctc gctcactgac tcgctgcgct
      541 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
      601 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
      661 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
      721 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
      781 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
      841 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt
      901 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
      961 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     1021 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     1081 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     1141 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     1201 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     1261 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     1321 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     1381 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     1441 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     1501 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     1561 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     1621 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     1681 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     1741 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     1801 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     1861 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     1921 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     1981 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     2041 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     2101 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     2161 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     2221 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     2281 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     2341 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     2401 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat
     2461 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     2521 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     2581 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     2641 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     2701 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     2761 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     2821 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     2881 taacgtttac aatttcccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     2941 tgcgggcctc ttcgctatta cgccagccca agctaccatg ataagtaagt aatattaagg
     3001 tacgggaggt acttggaggt acctagttat taatagtaat caattacggg gtcattagtt
     3061 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     3121 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     3181 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     3241 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     3301 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     3361 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
     3421 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     3481 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
     3541 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg
     3601 aaccgtcaga tccacgcgtg ccaccatgga caagaagtac tccattgggc tcgccatcgg
     3661 cacaaacagc gtcggctggg ccgtcattac ggacgagtac aaggtgccga gcaaaaaatt
     3721 caaagttctg ggcaataccg atcgccacag cataaagaag aacctcattg gcgccctcct
     3781 gttcgactcc ggggagacgg ccgaagccac gcggctcaaa agaacagcac ggcgcagata
     3841 tacccgcaga aagaatcgga tctgctacct gcaggagatc tttagtaatg agatggctaa
     3901 ggtggatgac tctttcttcc ataggctgga ggagtccttt ttggtggagg aggataaaaa
     3961 gcacgagcgc cacccaatct ttggcaatat cgtggacgag gtggcgtacc atgaaaagta
     4021 cccaaccata tatcatctga ggaagaagct tgtagacagt actgataagg ctgacttgcg
     4081 gttgatctat ctcgcgctgg cgcatatgat caaatttcgg ggacacttcc tcatcgaggg
     4141 ggacctgaac ccagacaaca gcgatgtcga caaactcttt atccaactgg ttcagactta
     4201 caatcagctt ttcgaagaga acccgatcaa cgcatccgga gttgacgcca aagcaatcct
     4261 gagcgctagg ctgtccaaat cccggcggct cgaaaacctc atcgcacagc tccctgggga
     4321 gaagaagaac ggcctgtttg gtaatcttat cgccctgtca ctcgggctga cccccaactt
     4381 taaatctaac ttcgacctgg ccgaagatgc caagcttcaa ctgagcaaag acacctacga
     4441 tgatgatctc gacaatctgc tggcccagat cggcgaccag tacgcagacc tttttttggc
     4501 ggcaaagaac ctgtcagacg ccattctgct gagtgatatt ctgcgagtga acacggagat
     4561 caccaaagct ccgctgagcg ctagtatgat caagcgctat gatgagcacc accaagactt
     4621 gactttgctg aaggcccttg tcagacagca actgcctgag aagtacaagg aaattttctt
     4681 cgatcagtct aaaaatggct acgccggata cattgacggc ggagcaagcc aggaggaatt
     4741 ttacaaattt attaagccca tcttggaaaa aatggacggc accgaggagc tgctggtaaa
     4801 gcttaacaga gaagatctgt tgcgcaaaca gcgcactttc gacaatggaa gcatccccca
     4861 ccagattcac ctgggcgaac tgcacgctat cctcaggcgg caagaggatt tctacccctt
     4921 tttgaaagat aacagggaaa agattgagaa aatcctcaca tttcggatac cctactatgt
     4981 aggccccctc gcccggggaa attccagatt cgcgtggatg actcgcaaat cagaagagac
     5041 catcactccc tggaacttcg aggaagtcgt ggataagggg gcctctgccc agtccttcat
     5101 cgaaaggatg actaactttg ataaaaatct gcctaacgaa aaggtgcttc ctaaacactc
     5161 tctgctgtac gagtacttca cagtttataa cgagctcacc aaggtcaaat acgtcacaga
     5221 agggatgaga aagccagcat tcctgtctgg agagcagaag aaagctatcg tggacctcct
     5281 cttcaagacg aaccggaaag ttaccgtgaa acagctcaaa gaagactatt tcaaaaagat
     5341 tgaatgtttc gactctgttg aaatcagcgg agtggaggat cgcttcaacg catccctggg
     5401 aacgtatcac gatctcctga aaatcattaa agacaaggac ttcctggaca atgaggagaa
     5461 cgaggacatt cttgaggaca ttgtcctcac ccttacgttg tttgaagata gggagatgat
     5521 tgaagaacgc ttgaaaactt acgctcatct cttcgacgac aaagtcatga aacagctcaa
     5581 gaggcgccga tatacaggat gggggcggct gtcaagaaaa ctgatcaatg ggatccgaga
     5641 caagcagagt ggaaagacaa tcctggattt tcttaagtcc gatggatttg ccaaccggaa
     5701 cttcatgcag ttgatccatg atgactctct cacctttaag gaggacatcc agaaagcaca
     5761 agtttctggc cagggggaca gtcttcacga gcacatcgct aatcttgcag gtagcccagc
     5821 tatcaaaaag ggaatactgc agaccgttaa ggtcgtggat gaactcgtca aagtaatggg
     5881 aaggcataag cccgagaata tcgttatcga gatggcccga gagaaccaaa ctacccagaa
     5941 gggacagaag aacagtaggg aaaggatgaa gaggattgaa gagggtataa aagaactggg
     6001 gtcccaaatc cttaaggaac acccagttga aaacacccag cttcagaatg agaagctcta
     6061 cctgtactac ctgcagaacg gcagggacat gtacgtggat caggaactgg acatcaatcg
     6121 gctctccgac tacgacgtgg atgccatcgt gccccagtct tttctcaaag atgattctat
     6181 tgataataaa gtgttgacaa gatccgataa aaatagaggg aagagtgata acgtcccctc
     6241 agaagaagtt gtcaagaaaa tgaaaaatta ttggcggcag ctgctgaacg ccaaactgat
     6301 cacacaacgg aagttcgata atctgactaa ggctgaacga ggtggcctgt ctgagttgga
     6361 taaagccggc ttcatcaaaa ggcagcttgt tgagacacgc cagatcacca agcacgtggc
     6421 ccaaattctc gattcacgca tgaacaccaa gtacgatgaa aatgacaaac tgattcgaga
     6481 ggtgaaagtt attactctga agtctaagct ggtctcagat ttcagaaagg actttcagtt
     6541 ttataaggtg agagagatca acaattacca ccatgcgcat gatgcctacc tgaatgcagt
     6601 ggtaggcact gcacttatca aaaaatatcc caagcttgaa tctgaatttg tttacggaga
     6661 ctataaagtg tacgatgtta ggaaaatgat cgcaaagtct gagcaggaaa taggcaaggc
     6721 caccgctaag tacttctttt acagcaatat tatgaatttt ttcaagaccg agattacact
     6781 ggccaatgga gagattcgga agcgaccact tatcgaaaca aacggagaaa caggagaaat
     6841 cgtgtgggac aagggtaggg atttcgcgac agtccggaag gtcctgtcca tgccgcaggt
     6901 gaacatcgtt aaaaagaccg aagtacagac cggaggcttc tccaaggaaa gtatcctccc
     6961 gaaaaggaac agcgacaagc tgatcgcacg caaaaaagat tgggacccca agaaatacgg
     7021 cggattcgat tctcctacag tcgcttacag tgtactggtt gtggccaaag tggagaaagg
     7081 gaagtctaaa aaactcaaaa gcgtcaagga actgctgggc atcacaatca tggagcgatc
     7141 aagcttcgaa aaaaacccca tcgactttct cgaggcgaaa ggatataaag aggtcaaaaa
     7201 agacctcatc attaagcttc ccaagtactc tctctttgag cttgaaaacg gccggaaacg
     7261 aatgctcgct agtgcgggcg agctgcagaa aggtaacgag ctggcactgc cctctaaata
     7321 cgttaatttc ttgtatctgg ccagccacta tgaaaagctc aaagggtctc ccgaagataa
     7381 tgagcagaag cagctgttcg tggaacaaca caaacactac cttgatgaga tcatcgagca
     7441 aataagcgaa ttctccaaaa gagtgatcct cgccgacgct aacctcgata aggtgctttc
     7501 tgcttacaat aagcacaggg ataagcccat cagggagcag gcagaaaaca ttatccactt
     7561 gtttactctg accaacttgg gcgcgcctgc agccttcaag tacttcgaca ccaccataga
     7621 cagaaagcgg tacacctcta caaaggaggt cctggacgcc acactgattc atcagtcaat
     7681 tacggggctc tatgaaacaa gaatcgacct ctctcagctc ggtggagaca gcagggctga
     7741 cgggccctca ctgggttcag ggtcacccaa gaagaaacgc aaagtcgagg atccaaagaa
     7801 gaaaaggaag gttgaagacc ccaagaaaaa gaggaaggtg gatgggatcg gctcaggcag
     7861 caacggcggt ggaggttcag acgctttgga cgatttcgat ctcgatatgc tcggttctga
     7921 cgccctggat gatttcgatc tggatatgct cggcagcgac gctctcgacg atttcgacct
     7981 cgacatgctc gggtcagatg ccttggatga ttttgacctg gatatgctct catgatgaac
     8041 tagtaccggt agatatcagc catggcttcc cgccggcggt ggcggcgcag gatgatggca
     8101 cgctgcccat gtcttgtgcc caggagagcg ggatggaccg tcaccctgca gcctgtgctt
     8161 ctgctaggat caatgtgtag gctagtcgag cagacatgat aagatacatt gatgagtttg
     8221 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     8281 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     8341 attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct
     8401 acaaatgtgg taaaatcgat aaggatccta ccgggtaggg gaggcgcttt tcccaaggca
     8461 gtctggagca tgcgctttag cagccccgct gggcacttgg cgctacacaa gtggcctctg
     8521 gcctcgcaca cattccacat ccaccggtag gcgccaaccg gctccgttct ttggtggccc
     8581 cttcgcgcca ccttctactc ctcccctagt caggaagttc ccccccgccc cgcagctcgc
     8641 gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact agtctcgtgc agatggacag
     8701 caccgctgag caatggaagc gggtaggcct ttggggcagc ggccaatagc agctttgctc
     8761 cttcgctttc tgggctcaga ggctgggaag gggtgggtcc gggggcgggc tcaggggcgg
     8821 gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc ccggcattct gcacgcttca
     8881 aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct ccgggccttt cgacctgcat
     8941 cccaagcttg ccaccatggt gtctaagggc gaagagctga ttaaggagaa catgcacatg
     9001 aagctgtaca tggagggcac cgtgaacaac caccacttca agtgcacatc cgagggcgaa
     9061 ggcaagccct acgagggcac ccagaccatg agaatcaagg tggtcgaggg cggccctctc
     9121 cccttcgcct tcgacatcct ggctaccagc ttcatgtacg gcagcaaaac cttcatcaac
     9181 cacacccagg gcatccccga cttctttaag cagtccttcc ctgagggctt cacatgggag
     9241 agagtcacca catacgaaga cgggggcgtg ctgaccgcta cccaggacac cagcctccag
     9301 gacggctgcc tcatctacaa cgtcaagatc agaggggtga acttcccatc caacggccct
     9361 gtgatgcaga agaaaacact cggctgggag gcctccaccg agatgctgta ccccgctgac
     9421 ggcggcctgg aaggcagaag cgacatggcc ctgaagctcg tgggcggggg ccacctgatc
     9481 tgcaacttga agaccacata cagatccaag aaacccgcta agaacctcaa gatgcccggc
     9541 gtctactatg tggacagaag actggaaaga atcaaggagg ccgacaaaga gacctacgtc
     9601 gagcagcacg aggtggctgt ggccagatac tgcgacctcc ctagcaaact ggggcacaaa
     9661 cttaattgac ccgggtaagc gggactctg
//
