LOCUS       sequence-438179-       12480 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses dCAS9-DNMT3A-DNMT3L fusion protein and double marker 
            GFP-T2A-Puro under independent promoters.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12480)
  TITLE     Systemic comparison of dCas9-based DNA methylation editing systems 
            for specificity and stability
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 12480)
  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            "Unpublished"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: M3-dCAS9-DNMT3A-DNMT3L (dCas9-3A-3L)
FEATURES             Location/Qualifiers
     source          1..12480
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(31..1614,1615..1617,1618..4104,4105..4107,
                     4108..4131))
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(31..4131)
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(4156..4176)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(4183..4248)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     regulatory      4248..4257
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          4269..4496
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     promoter        complement(4497..4773)
                     /label=chicken beta-actin promoter
     enhancer        4775..5060
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     misc_RNA        complement(5157..5232)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(5251..5499)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(5309..5328)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(5479..5499)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     rep_origin      complement(5561..6149)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5641..5660)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6320..7111,7112..7180))
                     /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     6943..6962
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7181..7285)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7353..7371
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7409..7431)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7531..7550
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7567..8022)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7704..7725)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7916..7935
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   8097..8237
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   8108..8237
                     /label=AAV2 ITR
     polyA_signal    8251..8372
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8262..8281)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     8316..8335
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(8540..9139)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APHNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     9120..9139
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(9140..9193)
                     /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             complement(join(9224..9934,9935..9937,9938..9940))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(9224..9934,9935..9937,9938..9940))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(9266..9287)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     9615..9634
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     regulatory      9937..9946
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(9985..10188)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(10014..10034)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        10189..10492
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    10507..10714
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     10703..10720
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(11503..12435)
                     /codon_start=1
                     /product="catalytic domain of DNA methyltransferase 3A"
                     /label=DNMT3A catalytic domain
                     /translation="PSRLQMFFANNHDQEFDPPKVYPPVPAEKRKPIRVLSLFDGIATG
                     LLVLKDLGIQVDRYIASEVCEDSITVGMVRHQGKIMYVGDVRSVTQKHIQEWGPFDLVI
                     GGSPCNDLSIVNPARKGLYEGTGRLFFEFYRLLHDARPKEGDDRPFFWLFENVVAMGVS
                     DKRDISRFLESNPVMIDAKEVSAAHRARYFWGNLPGMNRPLASTVNDKLELQECLEHGR
                     IAKFSKVRTITTRSNSIKQGKDQHFPVFMNEKEDILWCTEMERVFGFPVHYTDVSNMSR
                     LARQRLLGRSWSVPVIRHLFAPLKEYFACV"
     CDS             complement(join(12463..12480,1..30))
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
ORIGIN
        1 tccggccttt ttcgtggccg ccggcctttt gtcgcctccc agctgagaca ggtcgatccg
       61 tgtctcgtac aggccggtga tgctctggtg gatcagggtg gcgtccagca cctctttggt
      121 gctggtgtac ctcttccggt cgatggtggt gtcaaagtac ttgaaggcgg caggggctcc
      181 cagattggtc agggtaaaca ggtggatgat attctcggcc tgctctctga tgggcttatc
      241 ccggtgcttg ttgtaggcgg acagcacttt gtccagatta gcgtcggcca ggatcactct
      301 cttggagaac tcgctgatct gctcgatgat ctcgtccagg tagtgcttgt gctgttccac
      361 aaacagctgt ttctgctcat tatcctcggg ggagcccttc agcttctcat agtggctggc
      421 caggtacagg aagttcacat atttggaggg cagggccagt tcgtttccct tctgcagttc
      481 gccggcagag gccagcattc tcttccggcc gttttccagc tcgaacaggg agtacttagg
      541 cagcttgatg atcaggtcct ttttcacttc tttgtagccc ttggcttcca gaaagtcgat
      601 gggattcttc tcgaagctgc ttctttccat gatggtgatc cccagcagct ctttcacact
      661 cttcagtttc ttggacttgc ccttttccac tttggccacc accagcacag aataggccac
      721 ggtggggctg tcgaagccgc cgtacttctt agggtcccag tccttctttc tggcgatcag
      781 cttatcgctg ttcctcttgg gcaggataga ctctttgctg aagccgcctg tctgcacctc
      841 ggtctttttc acgatattca cttggggcat gctcagcact ttccgcacgg tggcaaaatc
      901 ccggccctta tcccacacga tctccccggt ttcgccgttt gtctcgatca gaggccgctt
      961 ccggatctcg ccgttggcca gggtaatctc ggtcttgaaa aagttcatga tgttgctgta
     1021 gaagaagtac ttggcggtag ccttgccgat ttcctgctcg ctcttggcga tcatcttccg
     1081 cacgtcgtac accttgtagt cgccgtacac gaactcgctt tccagcttag ggtacttttt
     1141 gatcagggcg gttcccacga cggcgttcag gtaggcgtcg tgggcgtggt ggtagttgtt
     1201 gatctcgcgc actttgtaaa actggaaatc cttccggaaa tcggacacca gcttggactt
     1261 cagggtgatc actttcactt cccggatcag cttgtcattc tcgtcgtact tagtgttcat
     1321 ccgggagtcc aggatctgtg ccacgtgctt tgtgatctgc cgggtttcca ccagctgtct
     1381 cttgatgaag ccggccttat ccagttcgct caggccgcct ctctcggcct tggtcagatt
     1441 gtcgaacttt ctctgggtaa tcagcttggc gttcagcagc tgccgccagt agttcttcat
     1501 cttcttcacg acctcttcgg agggcacgtt gtcgctcttg ccccggttct tgtcgcttct
     1561 ggtcagcacc ttgttgtcga tggagtcgtc cttcagaaag ctctgaggca cgatagcgtc
     1621 cacatcgtag tcggacagcc ggttgatgtc cagttcctgg tccacgtaca tatcccgccc
     1681 attctgcagg tagtacaggt acagcttctc gttctgcagc tgggtgtttt ccacggggtg
     1741 ttctttcagg atctggctgc ccagctcttt gatgccctct tcgatccgct tcattctctc
     1801 gcggctgttc ttctgtccct tctgggtggt ctggttctct ctggccattt cgatcacgat
     1861 gttctcgggc ttgtgccggc ccatcacttt cacgagctcg tccaccacct tcactgtctg
     1921 caggatgccc ttcttaatgg cggggctgcc ggccagattg gcaatgtgct cgtgcaggct
     1981 atcgccctgg ccggacacct gggctttctg gatgtcctct ttaaaggtca ggctgtcgtc
     2041 gtggatcagc tgcatgaagt ttctgttggc gaagccgtcg gacttcagga aatccaggat
     2101 tgtcttgccg gactgcttgt cccggatgcc gttgatcagc ttccggctca gcctgcccca
     2161 gccggtgtat ctccgccgct tcagctgctt catcactttg tcgtcgaaca ggtgggcata
     2221 ggttttcagc cgttcctcga tcatctctct gtcctcaaac agtgtcaggg tcagcacgat
     2281 atcttccaga atgtcctcgt tttcctcatt gtccaggaag tccttgtcct tgataatttt
     2341 cagcagatcg tggtatgtgc ccagggaggc gttgaaccga tcttccacgc cggagatttc
     2401 cacggagtcg aagcactcga ttttcttgaa gtagtcctct ttcagctgct tcacggtcac
     2461 tttccggttg gtcttgaaca gcaggtccac gatggccttt ttctgctcgc cgctcaggaa
     2521 ggcgggcttt ctcattccct cggtcacgta tttcactttg gtcagctcgt tatacacggt
     2581 gaagtactcg tacagcaggc tgtgcttggg cagcaccttc tcgttgggca ggttcttatc
     2641 gaagttggtc atccgctcga tgaagctctg ggcggaagcg cccttgtcca ccacttcctc
     2701 gaagttccag ggggtgatgg tttcctcgct ctttctggtc atccaggcga atctgctgtt
     2761 tcccctggcc agagggccca cgtagtaggg gatgcggaag gtcaggatct tctcgatctt
     2821 ttcccggttg tccttcagga atgggtaaaa atcttcctgc cgccgcagaa tggcgtgcag
     2881 ctctcccagg tggatctggt gggggatgct gccgttgtcg aaggtccgct gcttccgcag
     2941 caggtcctct ctgttcagct tcacgagcag ttcctcggtg ccgtccatct tttccaggat
     3001 gggcttgatg aacttgtaga actcttcctg gctggctccg ccgtcaatgt agccggcgta
     3061 gccgttcttg ctctggtcga agaaaatctc tttgtacttc tcaggcagct gctgccgcac
     3121 gagagctttc agcagggtca ggtcctggtg gtgctcgtcg tatctcttga tcatagaggc
     3181 gctcaggggg gccttggtga tctcggtgtt cactctcagg atgtcgctca gcaggatggc
     3241 gtcggacagg ttcttggcgg ccagaaacag gtcggcgtac tggtcgccga tctgggccag
     3301 caggttgtcc aggtcgtcgt cgtaggtgtc cttgctcagc tgcagtttgg catcctcggc
     3361 caggtcgaag ttgctcttga agttgggggt caggcccagg ctcagggcaa tcaggttgcc
     3421 gaacaggcca ttcttcttct cgccgggcag ctgggcgatc agattttcca gccgtctgct
     3481 cttgctcagt ctggcagaca ggatggcctt ggcgtccacg ccgctggcgt tgatggggtt
     3541 ttcctcgaac agctggttgt aggtctgcac cagctggatg aacagcttgt ccacgtcgct
     3601 gttgtcgggg ttcaggtcgc cctcgatcag gaagtggccc cggaacttga tcatgtgggc
     3661 cagggccaga tagatcagcc gcaggtcggc cttgtcggtg ctgtccacca gtttctttct
     3721 caggtggtag atggtggggt acttctcgtg gtaggccacc tcgtccacga tgttgccgaa
     3781 gatggggtgc cgctcgtgct tcttatcctc ttccaccagg aaggactctt ccagtctgtg
     3841 gaagaagctg tcgtccacct tggccatctc gttgctgaag atctcttgca gatagcagat
     3901 ccggttcttc cgtctggtgt atcttcttct ggcggttctc ttcagccggg tggcctcggc
     3961 tgtttcgccg ctgtcgaaca gcagggctcc gatcaggttc ttcttgatgc tgtgccggtc
     4021 ggtgttgccc agcaccttga atttcttgct gggcaccttg tactcgtcgg tgatcacggc
     4081 ccagcccaca gagttggtgc cgatggccag gccgatgctg tacttcttgt cggctgctgg
     4141 gactccgtgg ataccgacct tccgcttctt ctttggggcc atcttatcgt catcgtcttt
     4201 gtaatcaata tcatgatcct tgtagtctcc gtcgtggtcc ttatagtcca tggtggcacc
     4261 ggtccaacct gaaaaaaagt gatttcaggc aggtgctcca ggtaattaaa cattaatacc
     4321 ccaccaacca accatccctt aaacccttac ctcttgctca gctaattaca gcccggagga
     4381 gaagggccgt cccgcccgct cacctgtggg agtaacgcgg tcagtcagag ccggggcggg
     4441 cggcgcgagg cggcggcgga gcggggcacg gggcgaaggc agcgcgcagc gactcccgcc
     4501 cgccgcgcgc ttcgcttttt atagggccgc cgccgccgcc gcctcgccat aaaaggaaac
     4561 tttcggagcg cgccgctctg attggctgcc gccgcacctc tccgcctcgc cccgccccgc
     4621 ccctcgcccc gccccgcccc gcctggcgcg cgcccccccc cccccccgcc cccatcgctg
     4681 cacaaaataa ttaaaaaata aataaataca aaattggggg tggggagggg ggggagatgg
     4741 ggagagtgaa gcagaacgtg gggctcacct cgaccatggt aatagcgatg actaatacgt
     4801 agatgtactg ccaagtagga aagtcccata aggtcatgta ctgggcacaa tgccaggcgg
     4861 gccatttacc gtcattgacg tcaatagggg gcgtacttgg catatgatac acttgatgta
     4921 ctgccaagtg ggcagtttac cgtaaatact ccacccattg acgtcaatgg aaagtcccta
     4981 ttggcgttac tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat
     5041 ttaccgtaag ttatgtaacg ggtacctcta gagccatttg tctgcagaat tggcgcacgc
     5101 gctaaaaacg gactagcctt attttaactt gctatttcta gctctaaaac aaaaaagcac
     5161 cgactcggtg ccactttttc aagttgataa cggactagcc ttattttaac ttgctatttc
     5221 tagctctaaa acaggtcttc tcgaagaccc ggtgtttcgt cctttccaca agatatataa
     5281 agccaagaaa tcgaaatact ttcaagttac ggtaagcata tgatagtcca ttttaaaaca
     5341 taattttaaa actgcaaact acccaagaaa ttattacttt ctacgtcacg tattttgtac
     5401 taatatcttt gtgtttacag tcaaattaat tccaattatc tctctaacag ccttgtatcg
     5461 tatatgcaaa tatgaaggaa tcatgggaaa taggccctca catgtgagca aaaggccagc
     5521 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5581 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5641 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     5701 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     5761 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     5821 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     5881 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5941 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6001 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6061 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     6121 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6181 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6241 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6301 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6361 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6421 agggcttacc atctggcccc agtgctgcaa tgataccgcg gcttccacgc tcaccggctc
     6481 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6541 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6601 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     6661 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     6721 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     6781 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     6841 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     6901 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     6961 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     7021 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     7081 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     7141 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     7201 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7261 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     7321 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     7381 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     7441 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     7501 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     7561 accataaaat tgtaaacgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     7621 gctcattttt taaccaatag accgaaatcg gcaaaatccc ttataaatca aaagaatagc
     7681 ccgagataga gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     7741 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     7801 cacccaaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     7861 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     7921 agaaagcgaa aggagcgggc gctaaggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     7981 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtactatggt tgctttgacg
     8041 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgcccctg
     8101 caggcagctg cgcgctcgct cgctcactga ggccgcccgg gcaaagcccg ggcgtcgggc
     8161 gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc
     8221 catcactagg ggttcctgcg gccgccgcgt taagatacat tgatgagttt ggacaaacca
     8281 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     8341 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     8401 ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
     8461 gtatggctga ttatgatcag ttatctagat ccggtggatc ccgggcccgc ggtaccgtcg
     8521 actgcagaat tcgaagcttt caggcaccgg gcttgcgggt catgcaccag gtgcgcggtc
     8581 cttcgggcac ctcgacgtcg gcggtgacgg tgaagccgag ccgctcgtag aaggggaggt
     8641 tgtggggcgc ggaggtctcc aggaaggcgg gcaccccggc gcgctcggcc gcctccactc
     8701 cggggagcac gacggcgctg cccagaccct tgccctggtg gtcgggcgag acgccgacgg
     8761 tggccaggaa ccacgcgggc tccttgggcc ggtgcggcgc caggaggcct tccatctgtt
     8821 gctgcgcggc cagccgggaa ccgctcaact cggccatgcg cgggccgatc tcggcgaaca
     8881 ccgcccccgc ttcgacgctc tccggcgtgg tccagaccgc caccgcggcg ccgtcgtccg
     8941 cgacccacac cttgccgatg tcgagcccga cgcgcgtgag gaagagttct tgcagctcgg
     9001 tgacccgctc gatgtggcgg tccggatcga cggtgtggcg cgtggcgggg tagtcggcga
     9061 acgcggcggc gagggtgcgt acggccctgg ggacgtcgtc gcgggtggcg aggcgcaccg
     9121 tgggcttgta ctcggtcatt gggccaggat tctcctcgac gtcaccgcat gttagcagac
     9181 ttcctctgcc ctcagaagct tgagctcgag atctgagtcc ggacttgtac agctcgtcca
     9241 tgccgagagt gatcccggcg gcggtcacga actccagcag gaccatgtga tcgcgcttct
     9301 cgttggggtc tttgctcagg gcggactggg tgctcaggta gtggttgtcg ggcagcagca
     9361 cggggccgtc gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc acgctgccgt
     9421 cctcgatgtt gtggcggatc ttgaagttca ccttgatgcc gttcttctgc ttgtcggcca
     9481 tgatatagac gttgtggctg ttgtagttgt actccagctt gtgccccagg atgttgccgt
     9541 cctccttgaa gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact
     9601 tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgga
     9661 cgtagccttc gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc
     9721 ggctgaagca ctgcacgccg taggtcaggg tggtcacgag ggtgggccag ggcacgggca
     9781 gcttgccggt ggtgcagatg aacttcaggg tcagcttgcc gtaggtggca tcgccctcgc
     9841 cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg
     9901 gcaccacccc ggtgaacagc tcctcgccct tgctcaccat ggtggcgacc ggtagcgcta
     9961 gcggatctga cggttcacta aaccagctct gcttatatag acctcccacc gtacacgcct
    10021 accgcccatt tgcgtcaatg gggcggagtt gttacgacat tttggaaagt cccgttgatt
    10081 ttggtgccaa aacaaactcc cattgacgtc aatggggtgg agacttggaa atccccgtga
    10141 gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac cgcatcacca tggtaatagc
    10201 gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca tgtactgggc
    10261 acaatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac ttggcatatg
    10321 atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca
    10381 atggaaagtc cctattggcg ttactatggg aacatacgtc attattgacg tcaatgggcg
    10441 ggggtcgttg ggcggtcagc caggcgggcc atttaccgta agttatgtaa cgcggaacgc
    10501 ggccgctccc cagcatgcct gctattctct tcccaatcct cccccttgct gtcctgcccc
    10561 accccacccc ccagaataga atgacaccta ctcagacaat gcgatgcaat ttcctcattt
    10621 tattaggaaa ggacagtggg agtggcacct tccagggtca aggaaggcac gggggagggg
    10681 caaacaacag atggctggca actagaaggc acagtcgagg ctgatcagcg agctctagtt
    10741 agaattgaat tcgggccggc cctaaagagg aagtgagttt tgagaaaaat acttgaagta
    10801 ctctctcagc gggagaaggc agttcttcac caggaggtca actttcgggg cgtccagctt
    10861 gctcctgctt ctgacttggg cttgcagata ctcttcttcc tttggggtca ggggcgcatg
    10921 cttgctcttc agccctggaa tgttgctcca cacccgcata gcattctggt agtccctgcc
    10981 acggacatcc tggagggtca cagcctctgt ctgaaggaag cgggtagttg tctcttggtc
    11041 atcctcagtc agcagcagat tgtccatgaa tatccagaag aagggccgct gactctcctg
    11101 gcgaggcagc gcatactgca ggatccggtg gaactggaac atgtaccagc cgggacagcg
    11161 atcacaggag ctgcctaggg gctgcgtcga gccgtacacc aggtcaaagg ggccccattt
    11221 ctccacgtct ctcctcacga catttgtgac atcttccacg tacttcagcg ttcctccccc
    11281 agaaccagaa ccgctttcta aaaagcccaa actctttagt actttatcaa tatttctaaa
    11341 aaggctcagt acccgcactg gctgtctctt ccatgcagac actgtcttgt atatctccat
    11401 agggcccata tggctgccgc gcaagcttaa cggcaccagg ccgctgctga agctcggccc
    11461 gcggctgtta gcgttactat tgccgctaga aggccggccg gacacacacg caaaatactc
    11521 cttcagcgga gcgaagaggt ggcggatgac tggcacgctc catgaccggc ccagcagtct
    11581 ctgcctcgcc aagcggctca tgttggagac gtcagtatag tggactggga aaccaaatac
    11641 cctttccatt tcagtgcacc ataagatgtc ctctttctca ttcatgaaca caggaaaatg
    11701 ctggtctttg ccctgcttta tggagtttga cctcgtagta atggtcctca ctttgctgaa
    11761 cttggctatc ctgccatgct ccagacactc ctgcagctcc agcttatcat tcacagtgga
    11821 tgccaacggc ctgttcatac cgggaaggtt accccagaag tagcgggccc tgtgtgcagc
    11881 tgacacttct ttggcatcaa tcatcacagg gttggactcg agaaatcgcg agatgtccct
    11941 cttgtcacta acgcccatgg ccaccacatt ctcaaagagc cagaagaagg ggcgatcatc
    12001 tccctccttg ggccgcgcat catgcaggag gcggtagaac tcaaagaaga gccggccagt
    12061 gccctcgtag aggcccttgc gagcagggtt gacgatggag aggtcattgc agggactgcc
    12121 cccaatcacc agatcgaatg ggccccactc ctggatatgc ttctgtgtga cgctgcggac
    12181 gtccccgacg tacatgatct tcccctggtg ccgcaccatg cccaccgtga tggagtcctc
    12241 acacacctcc gaggcaatgt agcggtccac ctgaatgccc aagtccttca gcaccaggag
    12301 ccctgtagcg attccatcaa agagagacag cacccggatg ggcttcctct tctcagctgg
    12361 gacaggtggg taaacctttg gagggtcaaa ttcctggtcg tggttattag cgaagaacat
    12421 ctggagccgg gagggggatc cgctcccgcc tccgccctcg agctttttct ttttggcctg
//
