LOCUS       sequence-359690-       10657 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Fluorescently tagged dead Cas9 fused to the tripartite 
            transcriptional activator VP64-p65-Rta.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10657)
  AUTHORS   Erdel F, Rademacher A, Vlijm R, Tunnermann J, Frank L, Weinmann R, 
            Schweigert E, Yserentant K, Hummert J, Bauer C, Schumacher S, Al 
            Alwash A, Normand C, Herten DP, Engelhardt J, Rippe K
  TITLE     Mouse Heterochromatin Adopts Digital Compaction States without 
            Showing Hallmarks of HP1-Driven Liquid-Liquid Phase Separation.
  JOURNAL   Mol Cell. 2020 Apr 16;78(2):236-249.e7. doi: 
            10.1016/j.molcel.2020.02.005. Epub 2020 Feb 25.
  PUBMED    32101700
REFERENCE   2  (bases 1 to 10657)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.molcel.2020.02.005"; journalName: "Mol Cell"; date: 
            "2020-04-16- 16"; volume: "78"; issue: "2"; pages: "236-249.e7"
COMMENT     Sequence Label: pdCas9-GFP-VPR
FEATURES             Location/Qualifiers
     source          1..10657
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             628..648
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      697..706
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(703..729,730..732,733..3219,3220..3222,3223..4806)
                     /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             703..4806
                     /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             4810..4836
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4843..4863
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      4921..4930
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4927..4929,4930..4932,4933..5643)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4927..4929,4930..4932,4933..5643)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5233..5252)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5580..5601
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             5677..5697
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5764..5913
                     /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             5938..5958
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6388..6744
                     /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             6763..7332
                     /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    7467..7588
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7504..7523)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7558..7577
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(7595..8050)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7732..7753)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7944..7963
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        8077..8181
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        8183..8374
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(8206..8226)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      8371..8504
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             8553..9347
                     /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             8553..9347
                     /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(8607..8626)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     9217..9236
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(9535..9554)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    9579..9626
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      9955..10543
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10444..10463
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttactgggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgaggc caccatgccc aagaagaagc gcaaggtggg acgcgtctgc
      661 aggatatcaa gcttgcggta ccgcgggccc gggatcgcca ccatggacaa gaagtacagc
      721 atcggcctgg ccatcggcac caactctgtg ggctgggccg tgatcaccga cgagtacaag
      781 gtgcccagca agaaattcaa ggtgctgggc aacaccgacc ggcacagcat caagaagaac
      841 ctgatcggcg ccctgctgtt cgacagcgga gaaacagccg aggccacccg gctgaagaga
      901 accgccagaa gaagatacac cagacggaag aaccggatct gctatctgca agagatcttc
      961 agcaacgaga tggccaaggt ggacgacagc ttcttccaca gactggaaga gtccttcctg
     1021 gtggaagagg ataagaagca cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg
     1081 gcctaccacg agaagtaccc caccatctac cacctgagaa agaaactggt ggacagcacc
     1141 gacaaggccg acctgcggct gatctatctg gccctggccc acatgatcaa gttccggggc
     1201 cacttcctga tcgagggcga cctgaacccc gacaacagcg acgtggacaa gctgttcatc
     1261 cagctggtgc agacctacaa ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg
     1321 gacgccaagg ccatcctgtc tgccagactg agcaagagca gacggctgga aaatctgatc
     1381 gcccagctgc ccggcgagaa gaagaatggc ctgttcggca acctgattgc cctgagcctg
     1441 ggcctgaccc ccaacttcaa gagcaacttc gacctggccg aggatgccaa actgcagctg
     1501 agcaaggaca cctacgacga cgacctggac aacctgctgg cccagatcgg cgaccagtac
     1561 gccgacctgt ttctggccgc caagaacctg tccgacgcca tcctgctgag cgacatcctg
     1621 agagtgaaca ccgagatcac caaggccccc ctgagcgcct ctatgatcaa gagatacgac
     1681 gagcaccacc aggacctgac cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag
     1741 tacaaagaga ttttcttcga ccagagcaag aacggctacg ccggctacat cgatggcgga
     1801 gccagccagg aagagttcta caagttcatc aagcccatcc tggaaaagat ggacggcacc
     1861 gaggaactgc tcgtgaagct gaacagagag gacctgctgc ggaagcagcg gaccttcgac
     1921 aacggcagca tcccccacca gatccacctg ggagagctgc acgccattct gcggcggcag
     1981 gaagattttt acccattcct gaaggacaac cgggaaaaga tcgagaagat cctgaccttc
     2041 cgcatcccct actacgtggg ccctctggcc aggggaaaca gcagattcgc ctggatgacc
     2101 agaaagagcg aggaaaccat caccccctgg aacttcgagg aagtggtgga caagggcgcc
     2161 agcgcccaga gcttcatcga gcggatgacc aacttcgata agaacctgcc caacgagaag
     2221 gtgctgccca agcacagcct gctgtacgag tacttcaccg tgtacaacga gctgaccaaa
     2281 gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc tgagcggcga gcagaaaaaa
     2341 gccatcgtgg acctgctgtt caagaccaac cggaaagtga ccgtgaagca gctgaaagag
     2401 gactacttca agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg
     2461 ttcaacgcct ccctgggcac ataccacgat ctgctgaaaa ttatcaagga caaggacttc
     2521 ctggacaatg aggaaaacga ggacattctg gaagatatcg tgctgaccct gacactgttt
     2581 gaggacagag agatgatcga ggaacggctg aaaacctatg cccacctgtt cgacgacaaa
     2641 gtgatgaagc agctgaagcg gcggagatac accggctggg gcaggctgag ccggaagctg
     2701 atcaacggca tccgggacaa gcagtccggc aagacaatcc tggatttcct gaagtccgac
     2761 ggcttcgcca acagaaactt catgcagctg atccacgacg acagcctgac ctttaaagag
     2821 gacatccaga aagcccaggt gtccggccag ggcgatagcc tgcacgagca cattgccaat
     2881 ctggccggca gccccgccat taagaagggc atcctgcaga cagtgaaggt ggtggacgag
     2941 ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg tgatcgaaat ggccagagag
     3001 aaccagacca cccagaaggg acagaagaac agccgcgaga gaatgaagcg gatcgaagag
     3061 ggcatcaaag agctgggcag ccagatcctg aaagaacacc ccgtggaaaa cacccagctg
     3121 cagaacgaga agctgtacct gtactacctg cagaatgggc gggatatgta cgtggaccag
     3181 gaactggaca tcaaccggct gtccgactac gatgtggacg ctatcgtgcc tcagagcttt
     3241 ctgaaggacg actccatcga taacaaagtg ctgactcgga gcgacaagaa ccggggcaag
     3301 agcgacaacg tgccctccga agaggtcgtg aagaagatga agaactactg gcgccagctg
     3361 ctgaatgcca agctgattac ccagaggaag ttcgacaatc tgaccaaggc cgagagaggc
     3421 ggcctgagcg aactggataa ggccggcttc atcaagagac agctggtgga aacccggcag
     3481 atcacaaagc acgtggcaca gatcctggac tcccggatga acactaagta cgacgagaac
     3541 gacaaactga tccgggaagt gaaagtgatc accctgaagt ccaagctggt gtccgatttc
     3601 cggaaggatt tccagtttta caaagtgcgc gagatcaaca actaccacca cgcccacgac
     3661 gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc
     3721 gagttcgtgt acggcgacta caaggtgtac gacgtgcgga agatgatcgc caagagcgag
     3781 caggaaatcg gcaaggctac cgccaagtac ttcttctaca gcaacatcat gaactttttc
     3841 aagaccgaga ttaccctggc caacggcgag atccggaagc ggcctctgat cgagacaaac
     3901 ggcgaaacag gcgagatcgt gtgggataag ggccgggact ttgccaccgt gcggaaagtg
     3961 ctgtctatgc cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc
     4021 aaagagtcta tcctgcccaa gaggaacagc gacaagctga tcgccagaaa gaaggactgg
     4081 gaccctaaga agtacggcgg cttcgacagc cccaccgtgg cctattctgt gctggtggtg
     4141 gccaaagtgg aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct gctggggatc
     4201 accatcatgg aaagaagcag cttcgagaag aatcccatcg actttctgga agccaagggc
     4261 tacaaagaag tgaaaaagga cctgatcatc aagctgccta agtactccct gttcgagctg
     4321 gaaaacggcc ggaagagaat gctggcctct gccggcgaac tgcagaaggg aaacgaactg
     4381 gccctgccct ccaaatatgt gaacttcctg tacctggcca gccactatga gaagctgaag
     4441 ggctcccccg aggataatga gcagaaacag ctgtttgtgg aacagcacaa acactacctg
     4501 gacgagatca tcgagcagat cagcgagttc tccaagagag tgatcctggc cgacgctaat
     4561 ctggacaagg tgctgagcgc ctacaacaag cacagagaca agcctatcag agagcaggcc
     4621 gagaatatca tccacctgtt taccctgacc aatctgggag cccctgccgc cttcaagtac
     4681 tttgacacca ccatcgaccg gaagaggtac accagcacca aagaggtgct ggacgccacc
     4741 ctgatccacc agagcatcac cggcctgtac gagacacgga tcgacctgtc tcagctggga
     4801 ggcgacgcct atccctatga cgtgcccgat tatgcaccgg tcccaaaaaa gaaaagaaaa
     4861 gtaccggtcg ccacccgaat tctgcagtcg acggtaccgc gggcccggga tccaccggtc
     4921 gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     4981 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     5041 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     5101 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
     5161 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     5221 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     5281 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     5341 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag
     5401 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
     5461 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     5521 aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
     5581 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     5641 aagactagtc gcagggggcg tcgactgggt acccctccta agaagaagag gaaggttgcc
     5701 cccaccatgg ataaaaagaa gagaaaggcc cccccgaccg atgtcagcct gggggacgag
     5761 ctcgacgcat tggacgattt tgatctggat atgctgggaa gtgacgccct cgatgatttt
     5821 gaccttgaca tgcttggttc ggatgccctt gatgactttg acctcgacat gctcggcagt
     5881 gacgcccttg atgatttcga cctggacatg ctgattaact ctagaagttc cggatctccg
     5941 aaaaagaaac gcaaagttgg tagccagtac ctgcccgaca ccgacgaccg gcaccggatc
     6001 gaggaaaagc ggaagcggac ctacgagaca ttcaagagca tcatgaagaa gtcccccttc
     6061 agcggcccca ccgaccctag acctccacct agaagaatcg ccgtgcccag cagatccagc
     6121 gccagcgtgc caaaacctgc cccccagcct taccccttca ccagcagcct gagcaccatc
     6181 aactacgacg agttccctac catggtgttc cccagcggcc agatctctca ggcctctgct
     6241 ctggctccag cccctcctca ggtgctgcct caggctcctg ctcctgcacc agctccagcc
     6301 atggtgtctg cactggctca ggcaccagca cccgtgcctg tgctggctcc tggacctcca
     6361 caggctgtgg ctccaccagc ccctaaacct acacaggccg gcgagggcac actgtctgaa
     6421 gctctgctgc agctgcagtt cgacgacgag gatctgggag ccctgctggg aaacagcacc
     6481 gatcctgccg tgttcaccga cctggccagc gtggacaaca gcgagttcca gcagctgctg
     6541 aaccagggca tccctgtggc ccctcacacc accgagccca tgctgatgga ataccccgag
     6601 gccatcaccc ggctcgtgac aggcgctcag aggcctcctg atccagctcc tgcccctctg
     6661 ggagcaccag gcctgcctaa tggactgctg tctggcgacg aggacttcag ctctatcgcc
     6721 gatatggatt tctcagcctt gctgggctct ggcagcggca gccgggattc cagggaaggg
     6781 atgtttttgc cgaagcctga ggccggctcc gctattagtg acgtgtttga gggccgcgag
     6841 gtgtgccagc caaaacgaat ccggccattt catcctccag gaagtccatg ggccaaccgc
     6901 ccactccccg ccagcctcgc accaacacca accggtccag tacatgagcc agtcgggtca
     6961 ctgaccccgg caccagtccc tcagccactg gatccagcgc ccgcagtgac tcccgaggcc
     7021 agtcacctgt tggaggatcc cgatgaagag acgagccagg ctgtcaaagc ccttcgggag
     7081 atggccgata ctgtgattcc ccagaaggaa gaggctgcaa tctgtggcca aatggacctt
     7141 tcccatccgc ccccaagggg ccatctggat gagctgacaa ccacacttga gtccatgacc
     7201 gaggatctga acctggactc acccctgacc ccggaattga acgagattct ggataccttc
     7261 ctgaacgacg agtgcctctt gcatgccatg catatcagca caggactgtc catcttcgac
     7321 acatctctgt tttaaaacgt tcagagcggc cgcgactcta gatcataatc agccatacca
     7381 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     7441 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     7501 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     7561 gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta
     7621 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     7681 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     7741 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     7801 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     7861 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     7921 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     7981 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     8041 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     8101 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     8161 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     8221 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     8281 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     8341 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgccca
     8401 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     8461 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
     8521 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     8581 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     8641 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     8701 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     8761 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     8821 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     8881 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     8941 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     9001 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     9061 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
     9121 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     9181 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     9241 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     9301 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     9361 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     9421 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     9481 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
     9541 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     9601 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     9661 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     9721 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     9781 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta
     9841 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     9901 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     9961 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
    10021 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
    10081 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
    10141 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
    10201 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
    10261 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
    10321 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
    10381 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
    10441 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
    10501 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
    10561 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
    10621 ccctgattct gtggataacc gtattaccgc catgcat
//
