LOCUS       sequence-205527-       11128 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dCas9 and DNMT WT driven by pGK promoter and Puromycin driven by 
            EF1a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11128)
  AUTHORS   Pflueger C, Tan D, Swain T, Nguyen TV, Pflueger J, Nefzger C, Polo 
            JM, Ford E, Lister R
  TITLE     A modular dCas9-SunTag DNMT3A epigenome editing system overcomes 
            pervasive off-target activity of direct fusion dCas9-DNMT3A 
            constructs.
  JOURNAL   Genome Res. 2018 Jun 15. pii: gr.233049.117. doi: 
            10.1101/gr.233049.117.
  PUBMED    29907613
REFERENCE   2  (bases 1 to 11128)
  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: "Genome Res.
            2018 Jun 15. pii: gr.233049.117. doi: 10.1101/gr.233049.117."
COMMENT     Sequence Label: LLP249 pGK-dCas9-DNMT3a V3
FEATURES             Location/Qualifiers
     source          1..11128
                     /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"
     promoter        213..712
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(248..271)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     645..664
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             744..764
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             765..791
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             795..821
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             825..851
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(885..911,912..914,915..3398,3399..3404,3405..3470,
                     3471..3473,3474..4988)
                     /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(885..911,912..914,915..3401,3402..3404,3405..4988)
                     /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             885..4988
                     /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             5025..5045
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5085..5105
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5127..6059
                     /codon_start=1
                     /product="catalytic domain of DNA methyltransferase 3A"
                     /label=DNMT3A catalytic domain
                     /translation="PSRLQMFFANNHDQEFDPPKVYPPVPAEKRKPIRVLSLFDGIATG
                     LLVLKDLGIQVDRYIASEVCEDSITVGMVRHQGKIMYVGDVRSVTQKHIQEWGPFDLVI
                     GGSPCNDLSIVNPARKGLYEGTGRLFFEFYRLLHDARPKEGDDRPFFWLFENVVAMGVS
                     DKRDISRFLESNPVMIDAKEVSAAHRARYFWGNLPGMNRPLASTVNDKLELQECLEHGR
                     IAKFSKVRTITTRSNSIKQGKDQHFPVFMNEKEDILWCTEMERVFGFPVHYTDVSNMSR
                     LARQRLLGRSWSVPVIRHLFAPLKEYFACV"
     CDS             6159..6179
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(6208..6227)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6252..6299
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        6545..7723
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          6776..7714
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     7671..7691
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             7736..8335
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(7736..7755)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8232..8252
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     intron          8374..8603
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     8586..8605
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     polyA_signal    8748..8869
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8785..8804)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(8971..8987)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8971..8987)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8984..9006)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8995..9011
                     /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(9019..9025,9026..9043,9044..9049))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9064..9085
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9202..9219)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9373..9961)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9453..9472)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10132..10923,10924..10992))
                     /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     10755..10774
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10993..11097)
                     /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 ttaattaata ccgggtaggg gaggcgcttt tcccaaggca
      241 gtctggagca tgcgctttag cagccccgct gggcacttgg cgctacacaa gtggcctctg
      301 gcctcgcaca cattccacat ccaccggtag gcgccaaccg gctccgttct ttggtggccc
      361 cttcgcgcca ccttctactc ctcccctagt caggaagttc ccccccgccc cgcagctcgc
      421 gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact agtctcgtgc agatggacag
      481 caccgctgag caatggaagc gggtaggcct ttggggcagc ggccaatagc agctttgctc
      541 cttcgctttc tgggctcaga ggctgggaag gggtgggtcc gggggcgggc tcaggggcgg
      601 gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc ccggcattct gcacgcttca
      661 aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct ccgggccttt cggcggccgc
      721 acctgcagcc caagcttacc atgccaaaga agaagcggaa ggtctacccc tatgacgtgc
      781 ctgattacgc cgcttatcct tacgatgtgc ccgactatgc agcctaccca tacgatgtgc
      841 ccgactacgc cgcaatggcc ggtatccacg gagtcccagc agccatggac aagaagtact
      901 ccattgggct cgctatcggc acaaacagcg tcggctgggc cgtcattacg gacgagtaca
      961 aggtgccgag caaaaaattc aaagttctgg gcaataccga tcgccacagc ataaagaaga
     1021 acctcattgg cgccctcctg ttcgactccg gggagacggc cgaagccacg cggctcaaaa
     1081 gaacagcacg gcgcagatat acccgcagaa agaatcggat ctgctacctg caggagatct
     1141 ttagtaatga gatggctaag gtggatgact ctttcttcca taggctggag gagtcctttt
     1201 tggtggagga ggataaaaag cacgagcgcc acccaatctt tggcaatatc gtggacgagg
     1261 tggcgtacca tgaaaagtac ccaaccatat atcatctgag gaagaagctt gtagacagta
     1321 ctgataaggc tgacttgcgg ttgatctatc tcgcgctggc gcatatgatc aaatttcggg
     1381 gacacttcct catcgagggg gacctgaacc cagacaacag cgatgtcgac aaactcttta
     1441 tccaactggt tcagacttac aatcagcttt tcgaagagaa cccgatcaac gcatccggag
     1501 ttgacgccaa agcaatcctg agcgctaggc tgtccaaatc ccggcggctc gaaaacctca
     1561 tcgcacagct ccctggggag aagaagaacg gcctgtttgg taatcttatc gccctgtcac
     1621 tcgggctgac ccccaacttt aaatctaact tcgacctggc cgaagatgcc aagcttcaac
     1681 tgagcaaaga cacctacgat gatgatctcg acaatctgct ggcccagatc ggcgaccagt
     1741 acgcagacct ttttttggcg gcaaagaacc tgtcagacgc cattctgctg agtgatattc
     1801 tgcgagtgaa cacggagatc accaaagctc cgctgagcgc tagtatgatc aagcgctatg
     1861 atgagcacca ccaagacttg actttgctga aggcccttgt cagacagcaa ctgcctgaga
     1921 agtacaagga aattttcttc gatcagtcta aaaatggcta cgccggatac attgacggcg
     1981 gagcaagcca ggaggaattt tacaaattta ttaagcccat cttggaaaaa atggacggca
     2041 ccgaggagct gctggtaaag cttaacagag aagatctgtt gcgcaaacag cgcactttcg
     2101 acaatggaag catcccccac cagattcacc tgggcgaact gcacgctatc ctcaggcggc
     2161 aagaggattt ctaccccttt ttgaaagata acagggaaaa gattgagaaa atcctcacat
     2221 ttcggatacc ctactatgtg ggccccctcg cccggggaaa ttccagattc gcgtggatga
     2281 ctcgcaaatc agaagagacc atcactccct ggaacttcga ggaagtcgtg gataaggggg
     2341 cctctgccca gtccttcatc gaaaggatga ctaactttga taaaaatctg cctaacgaaa
     2401 aggtgcttcc taaacactct ctgctgtacg agtacttcac agtttataac gagctcacca
     2461 aggtcaaata cgtcacagaa gggatgagaa agccagcatt cctgtctgga gagcagaaga
     2521 aagctatcgt ggacctcctc ttcaagacga accggaaagt taccgtgaaa cagctcaaag
     2581 aagactattt caaaaagatt gaatgtttcg actctgttga aatcagcgga gtggaggatc
     2641 gcttcaacgc atccctggga acgtatcacg atctcctgaa aatcattaaa gacaaggact
     2701 tcctggacaa tgaggagaac gaggacattc ttgaggacat tgtcctcacc cttacgttgt
     2761 ttgaagatag ggagatgatt gaagaacgct tgaaaactta cgctcatctc ttcgacgaca
     2821 aagtcatgaa acagctcaag aggcgccgat atacaggatg ggggcggctg tcaagaaaac
     2881 tgatcaatgg gatccgagac aagcagagtg gaaagacaat cctggatttt cttaagtccg
     2941 atggatttgc caacaggaac ttcatgcagt tgatccatga tgactctctc acctttaagg
     3001 aggacatcca gaaagcacaa gtttctggcc agggggacag tcttcacgag cacatcgcta
     3061 atcttgcagg tagcccagct atcaaaaagg gaatactgca gaccgttaag gtcgtggatg
     3121 aactcgtcaa agtaatggga aggcataagc ccgagaatat cgttatcgag atggcccgag
     3181 agaaccaaac tacccagaag ggacagaaga acagtaggga aaggatgaag aggattgaag
     3241 agggtataaa agaactgggg tcccaaatcc ttaaggaaca cccagttgaa aacacccagc
     3301 ttcagaatga gaagctctac ctgtactacc tgcagaacgg cagggacatg tacgtggatc
     3361 aggaactgga catcaatcgg ctctccgact acgacgtgga tgcaatcgtg ccccagtctt
     3421 ttctcaaaga tgattctatt gataataaag tgttgacaag atccgataaa aatagaggga
     3481 agagtgataa cgtcccctca gaagaagttg tcaagaaaat gaaaaattat tggcggcagc
     3541 tgctgaacgc caaactgatc acacaacgga agttcgataa tctgactaag gctgaacgag
     3601 gtggcctgtc tgagttggat aaagccggct tcatcaaaag gcagcttgtt gagacacgcc
     3661 agatcaccaa gcacgtggcc caaattctcg attcacgcat gaacaccaag tacgatgaaa
     3721 atgacaaact gattcgagag gtgaaagtta ttactctgaa gtctaagctg gtctcagatt
     3781 tcagaaagga ctttcagttt tataaggtga gagagatcaa caattaccac catgcgcatg
     3841 atgcctacct gaatgcagtg gtaggcactg cacttatcaa aaaatatccc aagcttgaat
     3901 ctgaatttgt ttacggagac tataaagtgt acgatgttag gaaaatgatc gcaaagtctg
     3961 agcaggaaat aggcaaggcc accgctaagt acttctttta cagcaatatt atgaattttt
     4021 tcaagaccga gattacactg gccaatggag agattcggaa gcgaccactt atcgaaacaa
     4081 acggagaaac aggagaaatc gtgtgggaca agggtaggga tttcgcgaca gtccggaagg
     4141 tcctgtccat gccgcaggtg aacatcgtta aaaagaccga agtacagacc ggaggcttct
     4201 ccaaggaaag tatcctcccg aaaaggaaca gcgacaagct gatcgcacgc aaaaaagatt
     4261 gggaccccaa gaaatacggc ggattcgatt ctcctacagt cgcttacagt gtactggttg
     4321 tggccaaagt ggagaaaggg aagtctaaaa aactcaaaag cgtcaaggaa ctgctgggca
     4381 tcacaatcat ggagcgatca agcttcgaaa aaaaccccat cgactttctc gaggcgaaag
     4441 gatataaaga ggtcaaaaaa gacctcatca ttaagcttcc caagtactct ctctttgagc
     4501 ttgaaaacgg ccggaaacga atgctcgcta gtgcgggcga gctgcagaaa ggtaacgagc
     4561 tggcactgcc ctctaaatac gttaatttct tgtatctggc cagccactat gaaaagctca
     4621 aagggtctcc cgaagataat gagcagaagc agctgttcgt ggaacaacac aaacactacc
     4681 ttgatgagat catcgagcaa ataagcgaat tctccaaaag agtgatcctc gccgacgcta
     4741 acctcgataa ggtgctttct gcttacaata agcacaggga taagcccatc agggagcagg
     4801 cagaaaacat tatccacttg tttactctga ccaacttggg cgcgcctgca gccttcaagt
     4861 acttcgacac caccatagac agaaagcggt acacctctac aaaggaggtc ctggacgcca
     4921 cactgattca tcagtcaatt acggggctct atgaaacaag aatcgacctc tctcagctcg
     4981 gtggagacag cagggctgac gggggaggat cagggggagg gtcccccaaa aagaaacgca
     5041 aagtgaccgg caagaaaacc agcggccagg ccggacaggc ctctcctaag aaaaaaagaa
     5101 aagtgggacg cgctggcggc ggaagcccct cccggctcca gatgttcttc gctaataacc
     5161 acgaccagga atttgaccct ccaaaggttt acccacctgt cccagctgag aagaggaagc
     5221 ccatccgggt gctgtctctc tttgatggaa tcgctacagg gctcctggtg ctgaaggact
     5281 tgggcattca ggtggaccgc tacattgcct cggaggtgtg tgaggactcc atcacggtgg
     5341 gcatggtgcg gcaccagggg aagatcatgt acgtcgggga cgtccgcagc gtcacacaga
     5401 agcatatcca ggagtggggc ccattcgatc tggtgattgg gggcagtccc tgcaatgacc
     5461 tctccatcgt caaccctgct cgcaagggcc tctacgaggg cactggccgg ctcttctttg
     5521 agttctaccg cctcctgcat gatgcgcggc ccaaggaggg agatgatcgc cccttcttct
     5581 ggctctttga gaatgtggtg gccatgggcg ttagtgacaa gagggacatc tcgcgatttc
     5641 tcgagtccaa ccctgtgatg attgatgcca aagaagtgtc agctgcacac agggcccgct
     5701 acttctgggg taaccttccc ggtatgaaca ggccgttggc atccactgtg aatgataagc
     5761 tggagctgca ggagtgtctg gagcatggca ggatagccaa gttcagcaaa gtgaggacca
     5821 ttactacgag gtcaaactcc ataaagcagg gcaaagacca gcattttcct gtcttcatga
     5881 atgagaaaga ggacatctta tggtgcactg aaatggaaag ggtatttggt ttcccagtcc
     5941 actatactga cgtgtccaac atgagccgct tggcgaggca gagactgctg ggccggtcat
     6001 ggagcgtgcc agtcatccgc cacctcttcg ctccgctgaa ggagtatttt gcgtgtgtgg
     6061 aggtgcacac caaccaagat ccgcttgatg ctgaagttca tacaaaccag gatcccctcg
     6121 atgccgaagt ccatactaat caggacccac tggacgcacc caagaaaaag cgcaaagtcg
     6181 gcagcggcgc ctgaggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     6241 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
     6301 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     6361 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     6421 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagtgcaaa gatggataaa
     6481 gttttaaaca gagaggaatc tttgcagcta atggaccttc taggtcttga aaggagtggg
     6541 aattggctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt
     6601 ggggggaggg gtcggcaatt gaaccggtgc ctagagaagg tggcgcgggg taaactggga
     6661 aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag
     6721 tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacaggtaag
     6781 tgccgtgtgt ggttcccgcg ggcctggcct ctttacgggt tatggccctt gcgtgccttg
     6841 aattacttcc acctggctgc agtacgtgat tcttgatccc gagcttcggg ttggaagtgg
     6901 gtgggagagt tcgaggcctt gcgcttaagg agccccttcg cctcgtgctt gagttgaggc
     6961 ctggcctggg cgctggggcc gccgcgtgcg aatctggtgg caccttcgcg cctgtctcgc
     7021 tgctttcgat aagtctctag ccatttaaaa tttttgatga cctgctgcga cgcttttttt
     7081 ctggcaagat agtcttgtaa atgcgggcca agatctgcac actggtattt cggtttttgg
     7141 ggccgcgggc ggcgacgggg cccgtgcgtc ccagcgcaca tgttcggcga ggcggggcct
     7201 gcgagcgcgg ccaccgagaa tcggacgggg gtagtctcaa gctggccggc ctgctctggt
     7261 gcctggcctc gcgccgccgt gtatcgcccc gccctgggcg gcaaggctgg cccggtcggc
     7321 accagttgcg tgagcggaaa gatggccgct tcccggccct gctgcaggga gctcaaaatg
     7381 gaggacgcgg cgctcgggag agcgggcggg tgagtcaccc acacaaagga aaagggcctt
     7441 tccgtcctca gccgtcgctt catgtgactc cacggagtac cgggcgccgt ccaggcacct
     7501 cgattagttc tcgagctttt ggagtacgtc gtctttaggt tggggggagg ggttttatgc
     7561 gatggagttt ccccacactg agtgggtgga gactgaagtt aggccagctt ggcacttgat
     7621 gtaattctcc ttggaatttg ccctttttga gtttggatct tggttcattc tcaagcctca
     7681 gacagtggtt caaagttttt ttcttccatt tcaggtgtcg tgacgtacgg ccaccatgac
     7741 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtcccccggg ccgtacgcac
     7801 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgacc cggaccgcca
     7861 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg
     7921 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt
     7981 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg
     8041 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc
     8101 gtggttcctg gccaccgtcg gcgtgtcgcc cgaccaccag ggcaagggtc tgggcagcgc
     8161 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac
     8221 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt
     8281 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgatggaa
     8341 ttaattcgct gtctgcgagg gccagctgtt ggggtgagta ctccctctca aaagcgggca
     8401 tgacttctgc gctaagattg tcagtttcca aaaacgagga ggatttgata ttcacctggc
     8461 ccgcggtgat gcctttgagg gtggccgcgt ccatctggtc agaaaagaca atctttttgt
     8521 tgtcaagctt gaggtgtggc aggcttgaga tctggccata cacttgagtg acaatgacat
     8581 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caactgcagg tcgagataac
     8641 tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     8701 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     8761 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     8821 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa tgcgtcgagt
     8881 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     8941 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     9001 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     9061 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     9121 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     9181 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     9241 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     9301 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     9361 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     9421 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     9481 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     9541 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     9601 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     9661 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     9721 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     9781 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     9841 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     9901 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     9961 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
    10021 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
    10081 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
    10141 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
    10201 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
    10261 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
    10321 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
    10381 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
    10441 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
    10501 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
    10561 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
    10621 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
    10681 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
    10741 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
    10801 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
    10861 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
    10921 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
    10981 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
    11041 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
    11101 atttccccga aaagtgccac ctgacgtc
//
