LOCUS       sequence-313790-       11637 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Targeted CpG methylation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11637)
  AUTHORS   Lei Y, Zhang X, Su J, Jeong M, Gundry MC, Huang YH, Zhou Y, Li W, 
            Goodell MA
  TITLE     Targeted DNA methylation in vivo using an engineered dCas9-MQ1 
            fusion protein.
  JOURNAL   Nat Commun. 2017 Jul 11;8:16026. doi: 10.1038/ncomms16026.
  PUBMED    28695892
REFERENCE   2  (bases 1 to 11637)
  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: "Nat 
            Commun."; date: "2017-07-11"; volume: "8:16026. doi"; pages: " 
            10.1038/ncomms16026"
COMMENT     Sequence Label: pcDNA3.1-dCas9-MQ1(Q147L)-EGFP
FEATURES             Location/Qualifiers
     source          1..11637
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      754..763
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             763..828
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             835..855
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(865..891,892..894,895..3378,3379..3384,3385..3450,
                     3451..3453,3454..4968)
                     /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(865..891,892..894,895..3381,3382..3384,3385..4968)
                     /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             865..4968
                     /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             4981..5001
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6172..6225
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(6232..6234,6235..6945)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(6232..6234,6235..6945)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(6274..6295)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6535..6554)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6882..6903
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             6952..6969
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6992..7009)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6998..7222
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7268..7696
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7355..7374)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7565..7586
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7705..7725)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7710..8039
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7890..8025
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7952..7971
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             8106..8900
                     /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(8160..8179)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8770..8789
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    9076..9197
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9113..9132)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9167..9186
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9246..9262)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9246..9262)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9259..9281)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9270..9286
                     /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(9294..9300,9301..9318,9319..9324))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9339..9360
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9477..9494)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9648..10236)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9728..9747)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10407..11198,11199..11267))
                     /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     11030..11049
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11268..11372)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(11447..11466)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagttaag cttggtaccg agctcggatc cactagtcca gtgtggtgga attctgcaga
      721 tatccagcac agtggcggcc gctcgagtct agagccacca tggactacaa agaccatgac
      781 ggtgattata aagatcatga catcgattac aaggatgacg atgacaagat ggcccccaag
      841 aagaagagga aggtgggccg cggaatggac aagaagtact ccattgggct cgccatcggc
      901 acaaacagcg tcggctgggc cgtcattacg gacgagtaca aggtgccgag caaaaaattc
      961 aaagttctgg gcaataccga tcgccacagc ataaagaaga acctcattgg cgccctcctg
     1021 ttcgactccg gggaaaccgc cgaagccacg cggctcaaaa gaacagcacg gcgcagatat
     1081 acccgcagaa agaatcggat ctgctacctg caggagatct ttagtaatga gatggctaag
     1141 gtggatgact ctttcttcca taggctggag gagtcctttt tggtggagga ggataaaaag
     1201 cacgagcgcc acccaatctt tggcaatatc gtggacgagg tggcgtacca tgaaaagtac
     1261 ccaaccatat atcatctgag gaagaagctt gtagacagta ctgataaggc tgacttgcgg
     1321 ttgatctatc tcgcgctggc gcatatgatc aaatttcggg gacacttcct catcgagggg
     1381 gacctgaacc cagacaacag cgatgtcgac aaactcttta tccaactggt tcagacttac
     1441 aatcagcttt tcgaagagaa cccgatcaac gcatccggag ttgacgccaa agcaatcctg
     1501 agcgctaggc tgtccaaatc ccggcggctc gaaaacctca tcgcacagct ccctggggag
     1561 aagaagaacg gcctgtttgg taatcttatc gccctgtcac tcgggctgac ccccaacttt
     1621 aaatctaact tcgacctggc cgaagatgcc aagcttcaac tgagcaaaga cacctacgat
     1681 gatgatctcg acaatctgct ggcccagatc ggcgaccagt acgcagacct ttttttggcg
     1741 gcaaagaacc tgtcagacgc cattctgctg agtgatattc tgcgagtgaa cacggagatc
     1801 accaaagctc cgctgagcgc tagtatgatc aagcgctatg atgagcacca ccaagacttg
     1861 actttgctga aggcccttgt cagacagcaa ctgcctgaga agtacaagga aattttcttc
     1921 gatcagtcta aaaatggcta cgccggatac attgacggcg gagcaagcca ggaggaattt
     1981 tacaaattta ttaagcccat cttggaaaaa atggacggca ccgaggagct gctggtaaag
     2041 cttaacagag aagatctgtt gcgcaaacag cgcactttcg acaatggaag catcccccac
     2101 cagattcacc tgggcgaact gcacgctatc ctcaggcggc aagaggattt ctaccccttt
     2161 ttgaaagata acagggaaaa gattgagaaa atcctcacat ttcggatacc ctactatgta
     2221 ggccccctcg cccggggaaa ttccagattc gcgtggatga ctcgcaaatc agaagagacc
     2281 atcactccct ggaacttcga ggaagtcgtg gataaggggg cctctgccca gtccttcatc
     2341 gaaaggatga ctaactttga taaaaatctg cctaacgaaa aggtgcttcc taaacactct
     2401 ctgctgtacg agtacttcac agtttataac gagctcacca aggtcaaata cgtcacagaa
     2461 gggatgagaa agccagcatt cctgtctgga gagcagaaga aagctatcgt ggacctcctc
     2521 ttcaagacga accggaaagt taccgtgaaa cagctcaaag aagactattt caaaaagatt
     2581 gaatgtttcg actctgttga aatcagcgga gtggaggatc gcttcaacgc atccctggga
     2641 acgtatcacg atctcctgaa aatcattaaa gacaaggact tcctggacaa tgaggagaac
     2701 gaggacattc ttgaggacat tgtcctcacc cttacgttgt ttgaagatag ggagatgatt
     2761 gaagaacgct tgaaaactta cgctcatctc ttcgacgaca aagtcatgaa acagctcaag
     2821 aggcgccgat atacaggatg ggggcggctg tcaagaaaac tgatcaatgg gatccgagac
     2881 aagcagagtg gaaagacaat cctggatttt cttaagtccg atggatttgc caaccggaac
     2941 ttcatgcagt tgatccatga tgactctctc acctttaagg aggacatcca gaaagcacaa
     3001 gtttctggcc agggggacag tcttcacgag cacatcgcta atcttgcagg tagcccagct
     3061 atcaaaaagg gaatactgca gaccgttaag gtcgtggatg aactcgtcaa agtaatggga
     3121 aggcataagc ccgagaatat cgttatcgag atggcccgag agaaccaaac tacccagaag
     3181 ggacagaaga acagtaggga aaggatgaag aggattgaag agggtataaa agaactgggg
     3241 tcccaaatcc ttaaggaaca cccagttgaa aacacccagc ttcagaatga gaagctctac
     3301 ctgtactacc tgcagaacgg cagggacatg tacgtggatc aggaactgga catcaatcgg
     3361 ctctccgact acgacgtgga tgccatcgtg ccccagtctt ttctcaaaga tgattctatt
     3421 gataataaag tgttgacaag atccgataaa aatagaggga agagtgataa cgtcccctca
     3481 gaagaagttg tcaagaaaat gaaaaattat tggcggcagc tgctgaacgc caaactgatc
     3541 acacaacgga agttcgataa tctgactaag gctgaacgag gtggcctgtc tgagttggat
     3601 aaagccggct tcatcaaaag gcagcttgtt gagacacgcc agatcaccaa gcacgtggcc
     3661 caaattctcg attcacgcat gaacaccaag tacgatgaaa atgacaaact gattcgagag
     3721 gtgaaagtta ttactctraa gtctaagctg gtctcagatt tcagaaagga ctttcagttt
     3781 tataaggtga gagagatcaa caattaccac catgcgcatg atgcctacct gaatgcagtg
     3841 gtaggcactg cacttatcaa aaaatatccc aagcttgaat ctgaatttgt ttacggagac
     3901 tataaagtgt acgatgttag gaaaatgatc gcaaagtctg agcaggaaat aggcaaggcc
     3961 accgctaagt acttctttta cagcaatatt atgaattttt tcaagaccga gattacactg
     4021 gccaatggag agattcggaa gcgaccactt atcgaaacaa acggagaaac aggagaaatc
     4081 gtgtgggaca agggtaggga tttcgcgaca gtccggaagg tcctgtccat gccgcaggtg
     4141 aacatcgtta aaaagaccga agtacagacc ggaggcttct ccaaggaaag tatcctcccg
     4201 aaaaggaaca gcgacaagct gatcgcacgc aaaaaagatt gggaccccaa gaaatacggc
     4261 ggattcgatt ctcctacagt cgcttacagt gtactggttg tggccaaagt ggagaaaggg
     4321 aagtctaaaa aactcaaaag cgtcaaggaa ctgctgggca tcacaatcat ggagcgatca
     4381 agcttcgaaa aaaaccccat cgactttctc gaggcgaaag gatataaaga ggtcaaaaaa
     4441 gacctcatca ttaagcttcc caagtactct ctctttgagc ttgaaaacgg ccggaaacga
     4501 atgctcgcta gtgcgggcga gctgcagaaa ggtaacgagc tggcactgcc ctctaaatac
     4561 gttaatttct tgtatctggc cagccactat gaaaagctca aagggtctcc cgaagataat
     4621 gagcagaagc agctgttcgt ggaacaacac aaacactacc ttgatgagat catcgagcaa
     4681 ataagcgaat tctccaaaag agtgatcctc gccgacgcta acctcgataa ggtgctttct
     4741 gcttacaata agcacaggga taagcccatc agggagcagg cagaaaacat tatccacttg
     4801 tttactctga ccaacttggg cgcgcctgca gccttcaagt acttcgacac caccatagac
     4861 agaaagcggt acacctctac aaaggaggtc ctggacgcca cactgattca tcagtcaatt
     4921 acggggctct atgaaacaag aatcgacctc tctcagctcg gtggagacag cagggctgac
     4981 cccaagaaga agaggaaggt ggctagggac agcaaagtgg agaacaaaac aaagaagctg
     5041 agagtgttcg aagccttcgc cggcattggc gcccagagaa aggccctgga gaaagtgagg
     5101 aaggacgagt acgagatcgt gggactggcc gagtggtatg tgcccgccat cgtcatgtac
     5161 caggccatcc ataacaactt ccacaccaag ctcgagtaca agtccgtcag cagagaggag
     5221 atgatcgact acctcgagaa caagaccctg tcctggaaca gcaagaaccc cgtcagcaac
     5281 ggatactgga agaggaagaa ggatgacgag ctgaagatca tctacaacgc catcaagctg
     5341 tccgaaaagg agggcaacat tttcgacatc agggacctct acaagaggac actgaagaac
     5401 atcgacctgc tcacctacag cttcccttgc caggacctga gccagctagg catccagaag
     5461 ggcatgaaga ggggaagcgg caccagatcc ggcctgctct gggagatcga aagagccctg
     5521 gactccaccg agaagaacga cctgcctaag tacctcctca tggagaacgt gggagccctg
     5581 ctgcacaaga agaacgagga ggagctgaac caatggaagc agaagctgga gagcctgggc
     5641 taccagaaca gcatcgaagt cctcaatgct gccgatttcg gatccagcca ggccaggagg
     5701 agagtgttca tgatctccac cctcaatgag ttcgtggaac tgcctaaggg cgacaagaag
     5761 cccaagagca tcaaaaaggt gctgaacaag atcgtgagcg agaaggacat cctcaacaac
     5821 ctgctgaaat acaacctcac cgaattcaag aagaccaagt ccaacatcaa caaggccagc
     5881 ctgatcggct actccaagtt caactccgag ggctatgtgt acgaccccga gttcacaggc
     5941 cccacactga cagctagcgg cgccaactcc aggatcaaga tcaaggacgg cagcaacatc
     6001 aggaagatga acagcgacga gaccttcctg tacatcggct ttgacagcca ggacggcaag
     6061 agagtgaacg aaatcgagtt cctgaccgag aaccagaaga tcttcgtgtg tggcaacagc
     6121 atcagcgtgg aggtgctgga ggccatcatt gacaagatcg gcggccctag cgagggcaga
     6181 ggaagtcttc taacatgcgg tgacgtggag gagaatcccg gccctggtac cgtgagcaag
     6241 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     6301 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     6361 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     6421 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     6481 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     6541 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     6601 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     6661 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     6721 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     6781 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     6841 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     6901 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagaccgg tcatcatcac
     6961 catcaccatt gagtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     7021 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     7081 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     7141 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     7201 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     7261 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     7321 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     7381 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     7441 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     7501 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     7561 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     7621 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     7681 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     7741 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     7801 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     7861 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     7921 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     7981 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     8041 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
     8101 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     8161 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     8221 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     8281 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     8341 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     8401 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     8461 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     8521 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     8581 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     8641 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     8701 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     8761 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     8821 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     8881 ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga ccaagcgacg
     8941 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     9001 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     9061 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     9121 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     9181 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     9241 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     9301 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     9361 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     9421 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     9481 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     9541 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     9601 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     9661 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     9721 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     9781 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     9841 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     9901 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     9961 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
    10021 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
    10081 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
    10141 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
    10201 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
    10261 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
    10321 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
    10381 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
    10441 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
    10501 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
    10561 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
    10621 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
    10681 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
    10741 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
    10801 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
    10861 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
    10921 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
    10981 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
    11041 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
    11101 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
    11161 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
    11221 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
    11281 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
    11341 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
    11401 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
    11461 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
    11521 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
    11581 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgtt
//
