LOCUS       sequence-99786-e       11475 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  encodes S. pyogenes dCas9 with c-terminal fusion of human p300 HAT 
            core (aa 1048-1664) driven by CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11475)
  AUTHORS   Hilton IB, D'Ippolito AM, Vockley CM, Thakore PI, Crawford GE, Reddy
            TE, Gersbach CA
  TITLE     Epigenome editing by a CRISPR-Cas9-based acetyltransferase activates
            genes from promoters and enhancers.
  JOURNAL   Nat Biotechnol. 2015 May;33(5):510-7. doi: 10.1038/nbt.3199. Epub 
            2015 Apr 6.
  PUBMED    25849900
REFERENCE   2  (bases 1 to 11475)
  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"; doi: "10.1038/nbt.3199"; 
            journalName: "Nat Biotechnol"; date: "2015-05"; volume: "33"; issue:
            "5"; pages: "510-7"
COMMENT     Sequence Label: pcDNA-dCas9-p300 Core
FEATURES             Location/Qualifiers
     source          1..11475
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             907..972
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             979..999
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(1009..1035,1036..1038,1039..3525,3526..3528,
                     3529..5112)
                     /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             5137..5157
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5167..7017
                     /codon_start=1
                     /gene="EP300"
                     /product="catalytic histone acetyltransferase core of the 
                     human E1A-associated protein p300 (Delvecchio et al., 
                     2013)"
                     /label=p300 core
                     /translation="IFKPEELRQALMPTLEALYRQDPESLPFRQPVDPQLLGIPDYFDI
                     VKSPMDLSTIKRKLDTGQYQEPWQYVDDIWLMFNNAWLYNRKTSRVYKYCSKLSEVFEQ
                     EIDPVMQSLGYCCGRKLEFSPQTLCCYGKQLCTIPRDATYYSYQNRYHFCEKCFNEIQG
                     ESVSLGDDPSQPQTTINKEQFSKRKNDTLDPELFVECTECGRKMHQICVLHHEIIWPAG
                     FVCDGCLKKSARTRKENKFSAKRLPSTRLGTFLENRVNDFLRRQNHPESGEVTVRVVHA
                     SDKTVEVKPGMKARFVDSGEMAESFPYRTKALFAFEEIDGVDLCFFGMHVQEYGSDCPP
                     PNQRRVYISYLDSVHFFRPKCLRTAVYHEILIGYLEYVKKLGYTTGHIWACPPSEGDDY
                     IFHCHPPDQKIPKPKRLQEWYKKMLDKAVSERIVHDYKDIFKQATEDRLTSAKELPYFE
                     GDFWPNVLEESIKELEQEEEERKREENTSNESTDVTKGDSKNAKKKNNKKTSKNKSSLS
                     RGNKKKPGMPNVSNDLSQKLYATMEKHKEVFFVIRLIAGPAANSLPPIVDPDPLIPCDL
                     MDGRDAFLTLARDKHLEFSSLRRAQWSTMCMLVELHTQSQD"
     regulatory      6781..6790
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7018..7044
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(7069..7086)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7075..7299
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7345..7773
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7432..7451)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7642..7663
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7782..7802)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7787..8116
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7967..8102
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8029..8048
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             8183..8977
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(8237..8256)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8847..8866
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    9151..9272
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9188..9207)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9242..9261
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9321..9337)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9321..9337)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9334..9356)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9345..9361
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(9369..9375,9376..9393,9394..9399))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9414..9435
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9552..9569)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9723..10308)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9803..9822)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10479..11270,11271..11339))
                     /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     11102..11121
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11340..11444)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gccatggact acaaagacca tgacggtgat tataaagatc atgacatcga ttacaaggat
      961 gacgatgaca agatggcccc caagaagaag aggaaggtgg gccgcggaat ggataagaaa
     1021 tactcaatag gcttagctat cggcacaaat agcgtcggat gggcggtgat cactgatgaa
     1081 tataaggttc cgtctaaaaa gttcaaggtt ctgggaaata cagaccgcca cagtatcaaa
     1141 aaaaatctta taggggctct tttatttgac agtggagaga cagcggaagc gactcgtctc
     1201 aaacggacag ctcgtagaag gtatacacgt cggaagaatc gtatttgtta tctacaggag
     1261 attttttcaa atgagatggc gaaagtagat gatagtttct ttcatcgact tgaagagtct
     1321 tttttggtgg aagaagacaa gaagcatgaa cgtcatccta tttttggaaa tatagtagat
     1381 gaagttgctt atcatgagaa atatccaact atctatcatc tgcgaaaaaa attggtagat
     1441 tctactgata aagcggattt gcgcttaatc tatttggcct tagcgcatat gattaagttt
     1501 cgtggtcatt ttttgattga gggagattta aatcctgata atagtgatgt ggacaaacta
     1561 tttatccagt tggtacaaac ctacaatcaa ttatttgaag aaaaccctat taacgcaagt
     1621 ggagtagatg ctaaagcgat tctttctgca cgattgagta aatcaagacg attagaaaat
     1681 ctcattgctc agctccccgg tgagaagaaa aatggcttat ttgggaatct cattgctttg
     1741 tcattgggtt tgacccctaa ttttaaatca aattttgatt tggcagaaga tgctaaatta
     1801 cagctttcaa aagatactta cgatgatgat ttagataatt tattggcgca aattggagat
     1861 caatatgctg atttgttttt ggcagctaag aatttatcag atgctatttt actttcagat
     1921 atcctaagag taaatactga aataactaag gctcccctat cagcttcaat gattaaacgc
     1981 tacgatgaac atcatcaaga cttgactctt ttaaaagctt tagttcgaca acaacttcca
     2041 gaaaagtata aagaaatctt ttttgatcaa tcaaaaaacg gatatgcagg ttatattgat
     2101 gggggagcta gccaagaaga attttataaa tttatcaaac caattttaga aaaaatggat
     2161 ggtactgagg aattattggt gaaactaaat cgtgaagatt tgctgcgcaa gcaacggacc
     2221 tttgacaacg gctctattcc ccatcaaatt cacttgggtg agctgcatgc tattttgaga
     2281 agacaagaag acttttatcc atttttaaaa gacaatcgtg agaagattga aaaaatcttg
     2341 acttttcgaa ttccttatta tgttggtcca ttggcgcgtg gcaatagtcg ttttgcatgg
     2401 atgactcgga agtctgaaga aacaattacc ccatggaatt ttgaagaagt tgtcgataaa
     2461 ggtgcttcag ctcaatcatt tattgaacgc atgacaaact ttgataaaaa tcttccaaat
     2521 gaaaaagtac taccaaaaca tagtttgctt tatgagtatt ttacggttta taacgaattg
     2581 acaaaggtca aatatgttac tgaaggaatg cgaaaaccag catttctttc aggtgaacag
     2641 aagaaagcca ttgttgattt actcttcaaa acaaatcgaa aagtaaccgt taagcaatta
     2701 aaagaagatt atttcaaaaa aatagaatgt tttgatagtg ttgaaatttc aggagttgaa
     2761 gatagattta atgcttcatt aggtacctac catgatttgc taaaaattat taaagataaa
     2821 gattttttgg ataatgaaga aaatgaagat atcttagagg atattgtttt aacattgacc
     2881 ttatttgaag atagggagat gattgaggaa agacttaaaa catatgctca cctctttgat
     2941 gataaggtga tgaaacagct taaacgtcgc cgttatactg gttggggacg tttgtctcga
     3001 aaattgatta atggtattag ggataagcaa tctggcaaaa caatattaga ttttttgaaa
     3061 tcagatggtt ttgccaatcg caattttatg cagctgatcc atgatgatag tttgacattt
     3121 aaagaagaca ttcaaaaagc acaagtgtct ggacaaggcg atagtttaca tgaacatatt
     3181 gcaaatttag ctggtagccc tgctattaaa aaaggtattt tacagactgt aaaagttgtt
     3241 gatgaattgg tcaaagtaat ggggcggcat aagccagaaa atatcgttat tgaaatggca
     3301 cgtgaaaatc agacaactca aaagggccag aaaaattcgc gagagcgtat gaaacgaatc
     3361 gaagaaggta tcaaagaatt aggaagtcag attcttaaag agcatcctgt tgaaaatact
     3421 caattgcaaa atgaaaagct ctatctctat tatctccaaa atggaagaga catgtatgtg
     3481 gaccaagaat tagatattaa tcgtttaagt gattatgatg tcgatgccat tgttccacaa
     3541 agtttcctta aagacgattc aatagacaat aaggtcttaa cgcgttctga taaaaatcgt
     3601 ggtaaatcgg ataacgttcc aagtgaagaa gtagtcaaaa agatgaaaaa ctattggaga
     3661 caacttctaa acgccaagtt aatcactcaa cgtaagtttg ataatttaac gaaagctgaa
     3721 cgtggaggtt tgagtgaact tgataaagct ggttttatca aacgccaatt ggttgaaact
     3781 cgccaaatca ctaagcatgt ggcacaaatt ttggatagtc gcatgaatac taaatacgat
     3841 gaaaatgata aacttattcg agaggttaaa gtgattacct taaaatctaa attagtttct
     3901 gacttccgaa aagatttcca attctataaa gtacgtgaga ttaacaatta ccatcatgcc
     3961 catgatgcgt atctaaatgc cgtcgttgga actgctttga ttaagaaata tccaaaactt
     4021 gaatcggagt ttgtctatgg tgattataaa gtttatgatg ttcgtaaaat gattgctaag
     4081 tctgagcaag aaataggcaa agcaaccgca aaatatttct tttactctaa tatcatgaac
     4141 ttcttcaaaa cagaaattac acttgcaaat ggagagattc gcaaacgccc tctaatcgaa
     4201 actaatgggg aaactggaga aattgtctgg gataaagggc gagattttgc cacagtgcgc
     4261 aaagtattgt ccatgcccca agtcaatatt gtcaagaaaa cagaagtaca gacaggcgga
     4321 ttctccaagg agtcaatttt accaaaaaga aattcggaca agcttattgc tcgtaaaaaa
     4381 gactgggatc caaaaaaata tggtggtttt gatagtccaa cggtagctta ttcagtccta
     4441 gtggttgcta aggtggaaaa agggaaatcg aagaagttaa aatccgttaa agagttacta
     4501 gggatcacaa ttatggaaag aagttccttt gaaaaaaatc cgattgactt tttagaagct
     4561 aaaggatata aggaagttaa aaaagactta atcattaaac tacctaaata tagtcttttt
     4621 gagttagaaa acggtcgtaa acggatgctg gctagtgccg gagaattaca aaaaggaaat
     4681 gagctggctc tgccaagcaa atatgtgaat tttttatatt tagctagtca ttatgaaaag
     4741 ttgaagggta gtccagaaga taacgaacaa aaacaattgt ttgtggagca gcataagcat
     4801 tatttagatg agattattga gcaaatcagt gaattttcta agcgtgttat tttagcagat
     4861 gccaatttag ataaagttct tagtgcatat aacaaacata gagacaaacc aatacgtgaa
     4921 caagcagaaa atattattca tttatttacg ttgacgaatc ttggagctcc cgctgctttt
     4981 aaatattttg atacaacaat tgatcgtaaa cgatatacgt ctacaaaaga agttttagat
     5041 gccactctta tccatcaatc catcactggt ctttatgaaa cacgcattga tttgagtcag
     5101 ctaggaggtg acccaattgc cggatccaag gctagcccga aaaagaaacg caaagttggg
     5161 cgcgccattt tcaaaccaga agaactacga caggcactga tgccaacttt ggaggcactt
     5221 taccgtcagg atccagaatc ccttcccttt cgtcaacctg tggaccctca gcttttagga
     5281 atccctgatt actttgatat tgtgaagagc cccatggatc tttctaccat taagaggaag
     5341 ttagacactg gacagtatca ggagccctgg cagtatgtcg atgatatttg gcttatgttc
     5401 aataatgcct ggttatataa ccggaaaaca tcacgggtat acaaatactg ctccaagctc
     5461 tctgaggtct ttgaacaaga aattgaccca gtgatgcaaa gccttggata ctgttgtggc
     5521 agaaagttgg agttctctcc acagacactg tgttgctacg gcaaacagtt gtgcacaata
     5581 cctcgtgatg ccacttatta cagttaccag aacaggtatc atttctgtga gaagtgtttc
     5641 aatgagatcc aaggggagag cgtttctttg ggggatgacc cttcccagcc tcaaactaca
     5701 ataaataaag aacaattttc caagagaaaa aatgacacac tggatcctga actgtttgtt
     5761 gaatgtacag agtgcggaag aaagatgcat cagatctgtg tccttcacca tgagatcatc
     5821 tggcctgctg gattcgtctg tgatggctgt ttaaagaaaa gtgcacgaac taggaaagaa
     5881 aataagtttt ctgctaaaag gttgccatct accagacttg gcacctttct agagaatcgt
     5941 gtgaatgact ttctgaggcg acagaatcac cctgagtcag gagaggtcac tgttagagta
     6001 gttcatgctt ctgacaaaac cgtggaagta aaaccaggca tgaaagcaag gtttgtggac
     6061 agtggagaga tggcagaatc ctttccatac cgaaccaaag ccctctttgc ctttgaagaa
     6121 attgatggtg ttgacctgtg cttctttggc atgcatgttc aagagtatgg ctctgactgc
     6181 cctccaccca accagaggag agtatacata tcttacctcg atagtgttca tttcttccgt
     6241 cctaaatgct tgaggactgc agtctatcat gaaatcctaa ttggatattt agaatatgtc
     6301 aagaaattag gttacacaac agggcatatt tgggcatgtc caccaagtga gggagatgat
     6361 tatatcttcc attgccatcc tcctgaccag aagataccca agcccaagcg actgcaggaa
     6421 tggtacaaaa aaatgcttga caaggctgta tcagagcgta ttgtccatga ctacaaggat
     6481 atttttaaac aagctactga agatagatta acaagtgcaa aggaattgcc ttatttcgag
     6541 ggtgatttct ggcccaatgt tctggaagaa agcattaagg aactggaaca ggaggaagaa
     6601 gagagaaaac gagaggaaaa caccagcaat gaaagcacag atgtgaccaa gggagacagc
     6661 aaaaatgcta aaaagaagaa taataagaaa accagcaaaa ataagagcag cctgagtagg
     6721 ggcaacaaga agaaacccgg gatgcccaat gtatctaacg acctctcaca gaaactatat
     6781 gccaccatgg agaagcataa agaggtcttc tttgtgatcc gcctcattgc tggccctgct
     6841 gccaactccc tgcctcccat tgttgatcct gatcctctca tcccctgcga tctgatggat
     6901 ggtcgggatg cgtttctcac gctggcaagg gacaagcacc tggagttctc ttcactccga
     6961 agagcccagt ggtccaccat gtgcatgctg gtggagctgc acacgcagag ccaggactac
     7021 ccgtacgacg ttccggacta cgcttcttga gtttaaaccg ctgatcagcc tcgactgtgc
     7081 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     7141 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     7201 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     7261 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     7321 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     7381 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     7441 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     7501 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     7561 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     7621 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     7681 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     7741 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     7801 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     7861 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     7921 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     7981 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     8041 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     8101 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga
     8161 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     8221 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     8281 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     8341 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     8401 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     8461 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     8521 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     8581 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     8641 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     8701 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     8761 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     8821 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     8881 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     8941 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     9001 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     9061 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     9121 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     9181 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     9241 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     9301 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     9361 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     9421 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     9481 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     9541 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     9601 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     9661 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     9721 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     9781 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     9841 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     9901 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     9961 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
    10021 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
    10081 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
    10141 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
    10201 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
    10261 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
    10321 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
    10381 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
    10441 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
    10501 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
    10561 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
    10621 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
    10681 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
    10741 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
    10801 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
    10861 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
    10921 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
    10981 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
    11041 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
    11101 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
    11161 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
    11221 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
    11281 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
    11341 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
    11401 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
    11461 gtgccacctg acgtc
//
