LOCUS       sequence-481931-       17260 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes a CRISPR/dCas9-based fusion protein designed for targeted 
            DNA methylation..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 17260)
  TITLE     Versatile molecular tools enabling customizable DNA methylation in 
            Arabidopsis thaliana
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 17260)
  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: 
            "Unpublished"
COMMENT     Sequence Label: MQ1v-dCas9-BFP (M39)
FEATURES             Location/Qualifiers
     source          1..17260
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             3119..3139
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(3140..3166,3167..3169,3170..5656,5657..5659,
                     5660..7243)
                     /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             3140..7243
                     /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             7259..7279
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7283..7303
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7352..8050
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     terminator      8086..8338
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(8367..8383)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8367..8383)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8380..8402)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8391..8407
                     /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(8415..8421,8422..8439,8440..8445))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8460..8481
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8672..9348
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     primer_bind     9285..9305
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             9415..10440
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDDLMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     polyA_signal    10481..10655
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    10733..10757
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             11182..11976
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      12063..12651
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12552..12571
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12805..12822
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    12837..12977
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(12903..12922)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     13063..13085
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      13321..13515
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(13581..14654)
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     CDS             complement(15083..15712)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     misc_feature    17013..17037
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     17225..17247
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     17239..17256
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     17240..17256
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 agcttgcatg cctgcaggtc gactctagac ccggggatat attactcaac ttttgattcg
       61 ctatttgctg tgcacctgtg gcgttcatca catcttttgt gacactgttt gcactggtca
      121 ttgctattac aaaggacctt cctgatgttg aaggagatcg aaagtaagta actgcacgca
      181 taaccatttt ctttccgctc tttggctcaa tccatttgac agtcaaagac aatgtttaac
      241 cagctccgtt tgatatattg tctttatgtg tttgttcaag catgtttagt taatcatgcc
      301 tttgattgat cttgaatagg ttccaaatat caaccctggc aacaaaactt ggagtgagaa
      361 acattgcatt cctcggttct ggacttctgc tagtaaatta tgtttcagcc atatcactag
      421 ctttctacat gcctcaggtg aattcatcta tttccgtctt aactatttcg gttaattaaa
      481 gcacgaacac cattactgca tgtagaagct tgataaacta tcgccaccaa tttatttttg
      541 ttgcgatatt gttactttcc tcagtatgca gctttgaaaa gaccaaccct cttatccttt
      601 aacaatgaac aggtttttag aggtagcttg atgattcctg cacatgtgat cttggcttca
      661 ggcttaattt tccaggtaaa gcattatgag atactcttat atctcttaca tacttttgag
      721 ataatgcaca agaacttcat aactatatgc tttagtttct gcatttgaca ctgccaaatt
      781 cattaatctc taatatcttt gttgttgatc tttggtagac atgggtacta gaaaaagcaa
      841 actacaccaa ggtaaaatac ttttgtacaa acataaactc gttatcacgg aacatcaatg
      901 gagtgtatat ctaacggagt gtagaaacat ttgattattg caggaagcta tctcaggata
      961 ttatcggttt atatggaatc tcttctacgc agagtatctg ttattcccct tcctctagct
     1021 ttcaatttca tggtgaggat atgcagtttt ctttgtatat cattcttctt cttctttgta
     1081 gcttggagtc aaaatcggtt ccttcatgta catacatcaa ggatatgtcc ttctgaattt
     1141 ttatatcttg caataaaaat gcttgtacca attgaaacac cagctttttg agttctatga
     1201 tcactgactt ggttctaacc aaaaaaaaaa aaaatgttta atttacatat ctaaaagtag
     1261 gtttagggaa acctaaacag taaaatattt gtatattatt cgaatttcac tcatcataaa
     1321 aacttaaatt gcaccataaa attttgtttt actattaatg atgtaatttg tgtaacttaa
     1381 gataaaaata atattccgta agttaaccgg ctaaaaccac gtataaacca gggaacctgt
     1441 taaaccggtt ctttactgga taaagaaatg aaagcccatg tagacagctc cattagagcc
     1501 caaaccctaa atttctcatc tatataaaag gagtgacatt agggtttttg ttcgtcctct
     1561 taaagcttct cgttttctct gccgtctctc tcattcgcgc gacgcaaacg atcttcaggt
     1621 gatcttcttt ctccaaatcc tctctcataa ctctgatttc gtacttgtgt atttgagctc
     1681 acgctctgtt tctctcacca cagccatggc tagagattct aaggttgaga acaagactaa
     1741 gaaattgaga gttttcgaag ctttcgctgg aattggtgct cagagaaagg ctcttgagaa
     1801 agttagaaaa gatgagtatg aaatcgttgg attggctgaa tggtacgttc ctgctatcgt
     1861 tatgtaccaa gctatccata acaacttcca tactaagctt gagtacaagt ctgtttctag
     1921 agaagagatg atcgattacc ttgaaaacaa gacattgtct tggaactcta agaacccagt
     1981 ttctaacgga tactggaaga gaaagaaaga tgatgagctt aagatcatct acaacgctat
     2041 caagttgtct gagaaggagg gtaacatctt cgatatcaga gatctttaca agagaacttt
     2101 gaagaacatc gatcttttga catactcttt cccttgtcaa gatctttctc agttgggaat
     2161 ccaaaagggt atgaaaagag gatctggtac tagatctgga cttttgtggg aaattgaaag
     2221 agctcttgat tctacagaga agaacgattt gccaaagtac cttttgatgg aaaacgttgg
     2281 tgctcttttg cataagaaaa atgaagagga attgaaccaa tggaagcaga aacttgagtc
     2341 tttgggatac cagaattcta ttgaagttct taacgctgct gattttggtt cttctcaagc
     2401 tagaagaaga gttttcatga tctctactct taatgagttc gttgaattgc ctaagggtga
     2461 taagaagcca aagtctatca agaaagttct taacaagatc gtttctgaga aggatatcct
     2521 taacaacctt ttgaagtaca acttgactga attcaagaaa acaaagtcta acatcaacaa
     2581 ggcttctttg atcggatact ctaaatttaa ctctgagggt tatgtttacg atcctgaatt
     2641 cacaggacca actcttacag cttctggtgc taattctaga atcaagatca aggatggatc
     2701 taacatcaga aagatgaact ctgatgagac ttttctttac atcggattcg attctcagga
     2761 tggaaagaga gttaacgaga tcgaattctt gacagaaaac caaaagatct tcgtttgtgg
     2821 taactctatc tctgttgagg ttcttgaagc tattatcgat aaaatcggag gtccttctgg
     2881 agggccgagc tctggcgcac ccccaccaag tggagggtct cctgccgggt ccccaacatc
     2941 tactgaagaa ggcaccagcg aatccgcaac gcccgagtca ggccctggta catccacaga
     3001 accatctgaa ggtagtgcgc ctggttcccc agctggaagc cctacttcca ccgaagaagg
     3061 cacgtcaacc gaaccaagtg aaggatctgc ccctgggacc agcactgaac catctgagcc
     3121 aaaaaagaag agaaaggtaa tggacaagaa gtacagcatc ggcctggcca tcggcaccaa
     3181 ctctgtgggc tgggccgtga tcaccgacga gtacaaggtg cccagcaaga aattcaaggt
     3241 gctgggcaac accgaccggc acagcatcaa gaagaacctg atcggcgccc tgctgttcga
     3301 cagcggagaa acagccgagg ccacccggct gaagagaacc gccagaagaa gatacaccag
     3361 acggaagaac cggatctgct atctgcaaga gatcttcagc aacgagatgg ccaaggtgga
     3421 cgacagcttc ttccacagac tggaagagtc cttcctggtg gaagaggata agaagcacga
     3481 gcggcacccc atcttcggca acatcgtgga cgaggtggcc taccacgaga agtaccccac
     3541 catctaccac ctgagaaaga aactggtgga cagcaccgac aaggccgacc tgcggctgat
     3601 ctatctggcc ctggcccaca tgatcaagtt ccggggccac ttcctgatcg agggcgacct
     3661 gaaccccgac aacagcgacg tggacaagct gttcatccag ctggtgcaga cctacaacca
     3721 gctgttcgag gaaaacccca tcaacgccag cggcgtggac gccaaggcca tcctgtctgc
     3781 cagactgagc aagagcagac ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa
     3841 gaatggcctg ttcggcaacc tgattgccct gagcctgggc ctgaccccca acttcaagag
     3901 caacttcgac ctggccgagg atgccaaact gcagctgagc aaggacacct acgacgacga
     3961 cctggacaac ctgctggccc agatcggcga ccagtacgcc gacctgtttc tggccgccaa
     4021 gaacctgtcc gacgccatcc tgctgagcga catcctgaga gtgaacaccg agatcaccaa
     4081 ggcccccctg agcgcctcta tgatcaagag atacgacgag caccaccagg acctgaccct
     4141 gctgaaagct ctcgtgcggc agcagctgcc tgagaagtac aaagagattt tcttcgacca
     4201 gagcaagaac ggctacgccg gctacatcga tggcggagcc agccaggaag agttctacaa
     4261 gttcatcaag cccatcctgg aaaagatgga cggcaccgag gaactgctcg tgaagctgaa
     4321 cagagaggac ctgctgcgga agcagcggac cttcgacaac ggcagcatcc cccaccagat
     4381 ccacctggga gagctgcacg ccattctgcg gcggcaggaa gatttttacc cattcctgaa
     4441 ggacaaccgg gaaaagatcg agaagatcct gaccttccgc atcccctact acgtgggccc
     4501 tctggccagg ggaaacagca gattcgcctg gatgaccaga aagagcgagg aaaccatcac
     4561 cccctggaac ttcgaggaag tggtggacaa gggcgccagc gcccagagct tcatcgagcg
     4621 gatgaccaac ttcgataaga acctgcccaa cgagaaggtg ctgcccaagc acagcctgct
     4681 gtacgagtac ttcaccgtgt acaacgagct gaccaaagtg aaatacgtga ccgagggaat
     4741 gagaaagccc gccttcctga gcggcgagca gaaaaaagcc atcgtggacc tgctgttcaa
     4801 gaccaaccgg aaagtgaccg tgaagcagct gaaagaggac tacttcaaga aaatcgagtg
     4861 cttcgactcc gtggaaatct ccggcgtgga agatcggttc aacgcctccc tgggcacata
     4921 ccacgatctg ctgaaaatta tcaaggacaa ggacttcctg gacaatgagg aaaacgagga
     4981 cattctggaa gatatcgtgc tgaccctgac actgtttgag gacagagaga tgatcgagga
     5041 acggctgaaa acctatgccc acctgttcga cgacaaagtg atgaagcagc tgaagcggcg
     5101 gagatacacc ggctggggca ggctgagccg gaagctgatc aacggcatcc gggacaagca
     5161 gtccggcaag acaatcctgg atttcctgaa gtccgacggc ttcgccaaca gaaacttcat
     5221 gcagctgatc cacgacgaca gcctgacctt taaagaggac atccagaaag cccaggtgtc
     5281 cggccagggc gatagcctgc acgagcacat tgccaatctg gccggcagcc ccgccattaa
     5341 gaagggcatc ctgcagacag tgaaggtggt ggacgagctc gtgaaagtga tgggccggca
     5401 caagcccgag aacatcgtga tcgaaatggc cagagagaac cagaccaccc agaagggaca
     5461 gaagaacagc cgcgagagaa tgaagcggat cgaagagggc atcaaagagc tgggcagcca
     5521 gatcctgaaa gaacaccccg tggaaaacac ccagctgcag aacgagaagc tgtacctgta
     5581 ctacctgcag aatgggcggg atatgtacgt ggaccaggaa ctggacatca accggctgtc
     5641 cgactacgat gtggacgcta tcgtgcctca gagctttctg aaggacgact ccatcgataa
     5701 caaagtgctg actcggagcg acaagaaccg gggcaagagc gacaacgtgc cctccgaaga
     5761 ggtcgtgaag aagatgaaga actactggcg ccagctgctg aatgccaagc tgattaccca
     5821 gaggaagttc gacaatctga ccaaggccga gagaggcggc ctgagcgaac tggataaggc
     5881 cggcttcatc aagagacagc tggtggaaac ccggcagatc acaaagcacg tggcacagat
     5941 cctggactcc cggatgaaca ctaagtacga cgagaacgac aaactgatcc gggaagtgaa
     6001 agtgatcacc ctgaagtcca agctggtgtc cgatttccgg aaggatttcc agttttacaa
     6061 agtgcgcgag atcaacaact accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg
     6121 aaccgccctg atcaaaaagt accctaagct ggaaagcgag ttcgtgtacg gcgactacaa
     6181 ggtgtacgac gtgcggaaga tgatcgccaa gagcgagcag gaaatcggca aggctaccgc
     6241 caagtacttc ttctacagca acatcatgaa ctttttcaag accgagatta ccctggccaa
     6301 cggcgagatc cggaagcggc ctctgatcga gacaaacggc gaaacaggcg agatcgtgtg
     6361 ggataagggc cgggactttg ccaccgtgcg gaaagtgctg tctatgcccc aagtgaatat
     6421 cgtgaaaaag accgaggtgc agacaggcgg cttcagcaaa gagtctatcc tgcccaagag
     6481 gaacagcgac aagctgatcg ccagaaagaa ggactgggac cctaagaagt acggcggctt
     6541 cgacagcccc accgtggcct attctgtgct ggtggtggcc aaagtggaaa agggcaagtc
     6601 caagaaactg aagagtgtga aagagctgct ggggatcacc atcatggaaa gaagcagctt
     6661 cgagaagaat cccatcgact ttctggaagc caagggctac aaagaagtga aaaaggacct
     6721 gatcatcaag ctgcctaagt actccctgtt cgagctggaa aacggccgga agagaatgct
     6781 ggcctctgcc ggcgaactgc agaagggaaa cgaactggcc ctgccctcca aatatgtgaa
     6841 cttcctgtac ctggccagcc actatgagaa gctgaagggc tcccccgagg ataatgagca
     6901 gaaacagctg tttgtggaac agcacaaaca ctacctggac gagatcatcg agcagatcag
     6961 cgagttctcc aagagagtga tcctggccga cgctaatctg gacaaggtgc tgagcgccta
     7021 caacaagcac agagacaagc ctatcagaga gcaggccgag aatatcatcc acctgtttac
     7081 cctgaccaat ctgggagccc ctgccgcctt caagtacttt gacaccacca tcgaccggaa
     7141 gaggtacacc agcaccaaag aggtgctgga cgccaccctg atccaccaga gcatcaccgg
     7201 cctgtacgag acacggatcg acctgtctca gctgggaggc gacggtggcg gaggctcgcc
     7261 aaaaaagaag agaaaggtag acccaaagaa aaaacgaaaa gtaagcggaa gtgagacccc
     7321 aggtacatcc gaatcagcaa cgcctgaaag cagcgagctg attaaggaga acatgcacat
     7381 gaagctgtac atggagggca ccgtggacaa ccatcacttc aagtgcacat ccgagggcga
     7441 aggcaagccc tacgagggca cccagaccat gagaatcaag gtggtcgagg gcggccctct
     7501 ccccttcgcc ttcgacatcc tggctactag cttcctctac ggcagcaaga ccttcatcaa
     7561 ccacacccag ggcatccccg acttcttcaa gcagtccttc cctgagggct tcacatggga
     7621 gagagtcacc acatacgaag acgggggcgt gctgaccgct acccaggaca ccagcctcca
     7681 ggacggctgc ctcatctaca acgtcaagat cagaggggtg aacttcacat ccaacggccc
     7741 tgtgatgcag aagaaaacac tcggctggga ggccttcacc gagacgctgt accccgctga
     7801 cggcggcctg gaaggcagaa acgacatggc cctgaagctc gtgggcggga gccatctgat
     7861 cgcaaacatc aagaccacat atagatccaa gaaacccgct aagaacctca agatgcctgg
     7921 cgtctactat gtggactaca gactggaaag aatcaaggag gccaacaacg agacctacgt
     7981 cgagcagcac gaggtggcag tggccagata ctgcgacctc cctagcaaac tggggcacaa
     8041 gcttaattaa ggatcctgat tgatcgatag agctcgaatt tccccgatcg ttcaaacatt
     8101 tggcaataaa gtttcttaag attgaatcct gttgccggtc ttgcgatgat tatcatataa
     8161 tttctgttga attacgttaa gcatgtaata attaacatgt aatgcatgac gttatttatg
     8221 agatgggttt ttatgattag agtcccgcaa ttatacattt aatacgcgat agaaaacaaa
     8281 atatagcgcg caaactagga taaattatcg cgcgcggtgt catctatgtt actagatcgg
     8341 ggtaccgagc tcgaattcgt aatcatgtca tagctgtttc ctgtgtgaaa ttgttatccg
     8401 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     8461 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     8521 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     8581 ggctagagca gcttgccaac atggtggagc acgacactct cgtctactcc aagaatatca
     8641 aagatacagt ctcagaagac caaagggcta ttgagacttt tcaacaaagg gtaatatcgg
     8701 gaaacctcct cggattccat tgcccagcta tctgtcactt catcaaaagg acagtagaaa
     8761 aggaaggtgg cacctacaaa tgccatcatt gcgataaagg aaaggctatc gttcaagatg
     8821 cctctgccga cagtggtccc aaagatggac ccccacccac gaggagcatc gtggaaaaag
     8881 aagacgttcc aaccacgtct tcaaagcaag tggattgatg tgaacatggt ggagcacgac
     8941 actctcgtct actccaagaa tatcaaagat acagtctcag aagaccaaag ggctattgag
     9001 acttttcaac aaagggtaat atcgggaaac ctcctcggat tccattgccc agctatctgt
     9061 cacttcatca aaaggacagt agaaaaggaa ggtggcacct acaaatgcca tcattgcgat
     9121 aaaggaaagg ctatcgttca agatgcctct gccgacagtg gtcccaaaga tggaccccca
     9181 cccacgagga gcatcgtgga aaaagaagac gttccaacca cgtcttcaaa gcaagtggat
     9241 tgatgtgata tctccactga cgtaagggat gacgcacaat cccactatcc ttcgcaagac
     9301 ccttcctcta tataaggaag ttcatttcat ttggagagga cacgctgaaa tcaccagtct
     9361 ctctctacaa atctatctct ctcgagcttt cgcagatccg gggggcaatg agatatgaaa
     9421 aagcctgaac tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc
     9481 tccgacctga tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga
     9541 gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa agatcgttat
     9601 gtttatcggc actttgcatc ggccgcgctc ccgattccgg aagtgcttga cattggggag
     9661 tttagcgaga gcctgaccta ttgcatctcc cgccgttcac agggtgtcac gttgcaagac
     9721 ctgcctgaaa ccgaactgcc cgctgttcta caaccggtcg cggaggctat ggatgcgatc
     9781 gctgcggccg atcttagcca gacgagcggg ttcggcccat tcggaccgca aggaatcggt
     9841 caatacacta catggcgtga tttcatatgc gcgattgctg atccccatgt gtatcactgg
     9901 caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga tgatctgatg
     9961 ctttgggccg aggactgccc cgaagtccgg cacctcgtgc acgcggattt cggctccaac
    10021 aatgtcctga cggacaatgg ccgcataaca gcggtcattg actggagcga ggcgatgttc
    10081 ggggattccc aatacgaggt cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg
    10141 gagcagcaga cgcgctactt cgagcggagg catccggagc ttgcaggatc gccacgactc
    10201 cgggcgtata tgctccgcat tggtcttgac caactctatc agagcttggt tgacggcaat
    10261 ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg
    10321 actgtcgggc gtacacaaat cgcccgcaga agcgcggccg tctggaccga tggctgtgta
    10381 gaagtactcg ccgatagtgg aaaccgacgc cccagcactc gtccgagggc aaagaaatag
    10441 agtagatgcc gaccgggatc tgtcgatcga caagctcgag tttctccata ataatgtgtg
    10501 agtagttccc agataaggga attagggttc ctatagggtt tcgctcatgt gttgagcata
    10561 taagaaaccc ttagtatgta tttgtatttg taaaatactt ctatcaataa aatttctaat
    10621 tcctaaaacc aaaatccagt actaaaatcc agatcccccg aattaattcg gcgttaattc
    10681 agtacattaa aaacgtccgc aatgtgttat taagttgtct aagcgtcaat ttgtttacac
    10741 cacaatatat cctgccacca gccagccaac agctccccga ccggcagctc ggcacaaaat
    10801 caccactcga tacaggcagc ccatcagtcc gggacggcgt cagcgggaga gccgttgtaa
    10861 ggcggcagac tttgctcatg ttaccgatgc tattcggaag aacggcaact aagctgccgg
    10921 gtttgaaaca cggatgatct cgcggagggt agcatgttga ttgtaacgat gacagagcgt
    10981 tgctgcctgt gatcaccgcg gtttcaaaat cggctccgtc gatactatgt tatacgccaa
    11041 ctttgaaaac aactttgaaa aagctgtttt ctggtattta aggttttaga atgcaaggaa
    11101 cagtgaattg gagttcgtct tgttataatt agcttcttgg ggtatcttta aatactgtag
    11161 aaaagaggaa ggaaataata aatggctaaa atgagaatat caccggaatt gaaaaaactg
    11221 atcgaaaaat accgctgcgt aaaagatacg gaaggaatgt ctcctgctaa ggtatataag
    11281 ctggtgggag aaaatgaaaa cctatattta aaaatgacgg acagccggta taaagggacc
    11341 acctatgatg tggaacggga aaaggacatg atgctatggc tggaaggaaa gctgcctgtt
    11401 ccaaaggtcc tgcactttga acggcatgat ggctggagca atctgctcat gagtgaggcc
    11461 gatggcgtcc tttgctcgga agagtatgaa gatgaacaaa gccctgaaaa gattatcgag
    11521 ctgtatgcgg agtgcatcag gctctttcac tccatcgaca tatcggattg tccctatacg
    11581 aatagcttag acagccgctt agccgaattg gattacttac tgaataacga tctggccgat
    11641 gtggattgcg aaaactggga agaagacact ccatttaaag atccgcgcga gctgtatgat
    11701 tttttaaaga cggaaaagcc cgaagaggaa cttgtctttt cccacggcga cctgggagac
    11761 agcaacatct ttgtgaaaga tggcaaagta agtggcttta ttgatcttgg gagaagcggc
    11821 agggcggaca agtggtatga cattgccttc tgcgtccggt cgatcaggga ggatatcggg
    11881 gaagaacagt atgtcgagct attttttgac ttactgggga tcaagcctga ttgggagaaa
    11941 ataaaatatt atattttact ggatgaattg ttttagtacc tagaatgcat gaccaaaatc
    12001 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
    12061 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
    12121 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
    12181 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
    12241 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
    12301 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
    12361 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
    12421 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
    12481 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
    12541 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
    12601 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
    12661 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
    12721 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
    12781 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg
    12841 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc
    12901 agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg
    12961 actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
    13021 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
    13081 agaggttttc accgtcatca ccgaaacgcg cgaggcaggg tgccttgatg tgggcgccgg
    13141 cggtcgagtg gcgacggcgc ggcttgtccg cgccctggta gattgcctgg ccgtaggcca
    13201 gccatttttg agcggccagc ggccgcgata ggccgacgcg aagcggcggg gcgtagggag
    13261 cgcagcgacc gaagggtagg cgctttttgc agctcttcgg ctgtgcgctg gccagacagt
    13321 tatgcacagg ccaggcgggt tttaagagtt ttaataagtt ttaaagagtt ttaggcggaa
    13381 aaatcgcctt ttttctcttt tatatcagtc acttacatgt gtgaccggtt cccaatgtac
    13441 ggctttgggt tcccaatgta cgggttccgg ttcccaatgt acggctttgg gttcccaatg
    13501 tacgtgctat ccacaggaaa gagacctttt cgaccttttt cccctgctag ggcaatttgc
    13561 cctagcatct gctccgtaca ttaggaaccg gcggatgctt cgccctcgat caggttgcgg
    13621 tagcgcatga ctaggatcgg gccagcctgc cccgcctcct ccttcaaatc gtactccggc
    13681 aggtcatttg acccgatcag cttgcgcacg gtgaaacaga acttcttgaa ctctccggcg
    13741 ctgccactgc gttcgtagat cgtcttgaac aaccatctgg cttctgcctt gcctgcggcg
    13801 cggcgtgcca ggcggtagag aaaacggccg atgccgggat cgatcaaaaa gtaatcgggg
    13861 tgaaccgtca gcacgtccgg gttcttgcct tctgtgatct cgcggtacat ccaatcagct
    13921 agctcgatct cgatgtactc cggccgcccg gtttcgctct ttacgatctt gtagcggcta
    13981 atcaaggctt caccctcgga taccgtcacc aggcggccgt tcttggcctt cttcgtacgc
    14041 tgcatggcaa cgtgcgtggt gtttaaccga atgcaggttt ctaccaggtc gtctttctgc
    14101 tttccgccat cggctcgccg gcagaacttg agtacgtccg caacgtgtgg acggaacacg
    14161 cggccgggct tgtctccctt cccttcccgg tatcggttca tggattcggt tagatgggaa
    14221 accgccatca gtaccaggtc gtaatcccac acactggcca tgccggccgg ccctgcggaa
    14281 acctctacgt gcccgtctgg aagctcgtag cggatcacct cgccagctcg tcggtcacgc
    14341 ttcgacagac ggaaaacggc cacgtccatg atgctgcgac tatcgcgggt gcccacgtca
    14401 tagagcatcg gaacgaaaaa atctggttgc tcgtcgccct tgggcggctt cctaatcgac
    14461 ggcgcaccgg ctgccggcgg ttgccgggat tctttgcgga ttcgatcagc ggccgcttgc
    14521 cacgattcac cggggcgtgc ttctgcctcg atgcgttgcc gctgggcggc ctgcgcggcc
    14581 ttcaacttct ccaccaggtc atcacccagc gccgcgccga tttgtaccgg gccggatggt
    14641 ttgcgaccgc tcacgccgat tcctcgggct tgggggttcc agtgccattg cagggccggc
    14701 aggcaaccca gccgcttacg cctggccaac cgcccgttcc tccacacatg gggcattcca
    14761 cggcgtcggt gcctggttgt tcttgatttt ccatgccgcc tcctttagcc gctaaaattc
    14821 atctactcat ttattcattt gctcatttac tctggtagct gcgcgatgta ttcagatagc
    14881 agctcggtaa tggtcttgcc ttggcgtacc gcgtacatct tcagcttggt gtgatcctcc
    14941 gccggcaact gaaagttgac ccgcttcatg gctggcgtgt ctgccaggct ggccaacgtt
    15001 gcagccttgc tgctgcgtgc gctcggacgg ccggcactta gcgtgtttgt gcttttgctc
    15061 attttctctt tacctcatta actcaaatga gttttgattt aatttcagcg gccagcgcct
    15121 ggacctcgcg ggcagcgtcg ccctcgggtt ctgattcaag aacggttgtg ccggcggcgg
    15181 cagtgcctgg gtagctcacg cgctgcgtga tacgggactc aagaatgggc agctcgtacc
    15241 cggccagcgc ctcggcaacc tcaccgccga tgcgcgtgcc tttgatcgcc cgcgacacga
    15301 caaaggccgc ttgtagcctt ccatccgtga cctcaatgcg ctgcttaacc agctccacca
    15361 ggtcggcggt ggcccatatg tcgtaagggc ttggctgcac cggaatcagc acgaagtcgg
    15421 ctgccttgat cgcggacaca gccaagtccg ccgcctgggg cgctccgtcg atcactacga
    15481 agtcgcgccg gccgatggcc ttcacgtcgc ggtcaatcgt cgggcggtcg atgccgacaa
    15541 cggttagcgg ttgatcttcc cgcacggccg cccaatcgcg ggcactgccc tggggatcgg
    15601 aatcgactaa cagaacatcg gccccggcga gttgcagggc gcgggctaga tgggttgcga
    15661 tggtcgtctt gcctgacccg cctttctggt taagtacagc gataaccttc atgcgttccc
    15721 cttgcgtatt tgtttattta ctcatcgcat catatacgca gcgaccgcat gacgcaagct
    15781 gttttactca aatacacatc acctttttag acggcggcgc tcggtttctt cagcggccaa
    15841 gctggccggc caggccgcca gcttggcatc agacaaaccg gccaggattt catgcagccg
    15901 cacggttgag acgtgcgcgg gcggctcgaa cacgtacccg gccgcgatca tctccgcctc
    15961 gatctcttcg gtaatgaaaa acggttcgtc ctggccgtcc tggtgcggtt tcatgcttgt
    16021 tcctcttggc gttcattctc ggcggccgcc agggcgtcgg cctcggtcaa tgcgtcctca
    16081 cggaaggcac cgcgccgcct ggcctcggtg ggcgtcactt cctcgctgcg ctcaagtgcg
    16141 cggtacaggg tcgagcgatg cacgccaagc agtgcagccg cctctttcac ggtgcggcct
    16201 tcctggtcga tcagctcgcg ggcgtgcgcg atctgtgccg gggtgagggt agggcggggg
    16261 ccaaacttca cgcctcgggc cttggcggcc tcgcgcccgc tccgggtgcg gtcgatgatt
    16321 agggaacgct cgaactcggc aatgccggcg aacacggtca acaccatgcg gccggccggc
    16381 gtggtggtgt cggcccacgg ctctgccagg ctacgcaggc ccgcgccggc ctcctggatg
    16441 cgctcggcaa tgtccagtag gtcgcgggtg ctgcgggcca ggcggtctag cctggtcact
    16501 gtcacaacgt cgccagggcg taggtggtca agcatcctgg ccagctccgg gcggtcgcgc
    16561 ctggtgccgg tgatcttctc ggaaaacagc ttggtgcagc cggccgcgtg cagttcggcc
    16621 cgttggttgg tcaagtcctg gtcgtcggtg ctgacgcggg catagcccag caggccagcg
    16681 gcggcgctct tgttcatggc gtaatgtctc cggttctagt cgcaagtatt ctactttatg
    16741 cgactaaaac acgcgacaag aaaacgccag gaaaagggca gggcggcagc ctgtcgcgta
    16801 acttaggact tgtgcgacat gtcgttttca gaagacggct gcactgaacg tcagaagccg
    16861 actgcactat agcagcggag gggttggatc aaagtacttt gatcccgagg ggaaccctgt
    16921 ggttggcatg cacatacaaa tggacgaacg gataaacctt ttcacgccct tttaaatatc
    16981 cgattattct aataaacgct cttttctctt aggtttaccc gccaatatat cctgtcaaac
    17041 actgatagtt taaactgaag gcgggaaacg acaatctgat ccaagctcaa gctgctctag
    17101 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
    17161 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
    17221 ttttcccagt cacgacgttg taaaacgacg gccagtgcca
//
