LOCUS       sequence-438195-       11743 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses dCAS9-dead DNMT3A 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 11743)
  TITLE     Systemic comparison of dCas9-based DNA methylation editing systems 
            for specificity and stability
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 11743)
  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            "Unpublished"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-dDNMT3A (dCas9-d3A)
FEATURES             Location/Qualifiers
     source          1..11743
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(3889..3909)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(3916..3981)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     regulatory      3981..3990
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          4002..4229
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     promoter        complement(4230..4507)
                     /label=chicken beta-actin promoter
     enhancer        4509..4794
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     misc_RNA        complement(4891..4966)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(4985..5233)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(5043..5062)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(5213..5233)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     rep_origin      complement(5295..5883)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5375..5394)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6054..6845,6846..6914))
                     /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     6677..6696
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6915..7019)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7087..7105
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7143..7165)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7265..7284
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7301..7756)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7438..7459)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7650..7669
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   7831..7971
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   7842..7971
                     /label=AAV2 ITR
     polyA_signal    7985..8106
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7996..8015)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     8050..8069
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(8274..8873)
                     /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     8854..8873
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(8874..8927)
                     /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(8958..9668,9669..9671,9672..9674))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(8958..9668,9669..9671,9672..9674))
                     /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(9000..9021)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     9349..9368
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     regulatory      9671..9680
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(9719..9922)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(9748..9768)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        9923..10226
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    10241..10448
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     10437..10454
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(10499..11431)
                     /codon_start=1
                     /product="catalytic domain of DNA methyltransferase 3A"
                     /label=DNMT3A catalytic domain
                     /translation="PSRLQMFFANNHDQEFDPPKVYPPVPAEKRKPIRVLSLFDGIATG
                     LLVLKDLGIQVDRYIASEVCEDSITVGMVRHQGKIMYVGDVRSVTQKHIQEWGPFDLVI
                     GGSPCNDLSIVNPARKGLYEGTGRLFFEFYRLLHDARPKEGDDRPFFWLFANVVAMGVS
                     DKRDISRFLESNPVMIDAKEVSAAHRARYFWGNLPGMNRPLASTVNDKLELQECLEHGR
                     IAKFSKVRTITTRSNSIKQGKDQHFPVFMNEKEDILWCTEMERVFGFPVHYTDVSNMSR
                     LARQRLLGRSWSVPVIRHLFAPLKEYFACV"
     CDS             complement(11459..11506)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(join(11507..11743,1..1347,1348..1350,1351..3837,
                     3838..3840,3841..3864))
                     /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(join(11507..11743,1..3864))
                     /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"
ORIGIN
        1 tttgtccaga ttagcgtcgg ccaggatcac tctcttggag aactcgctga tctgctcgat
       61 gatctcgtcc aggtagtgct tgtgctgttc cacaaacagc tgtttctgct cattatcctc
      121 gggggagccc ttcagcttct catagtggct ggccaggtac aggaagttca catatttgga
      181 gggcagggcc agttcgtttc ccttctgcag ttcgccggca gaggccagca ttctcttccg
      241 gccgttttcc agctcgaaca gggagtactt aggcagcttg atgatcaggt cctttttcac
      301 ttctttgtag cccttggctt ccagaaagtc gatgggattc ttctcgaagc tgcttctttc
      361 catgatggtg atccccagca gctctttcac actcttcagt ttcttggact tgcccttttc
      421 cactttggcc accaccagca cagaataggc cacggtgggg ctgtcgaagc cgccgtactt
      481 cttagggtcc cagtccttct ttctggcgat cagcttatcg ctgttcctct tgggcaggat
      541 agactctttg ctgaagccgc ctgtctgcac ctcggtcttt ttcacgatat tcacttgggg
      601 catgctcagc actttccgca cggtggcaaa atcccggccc ttatcccaca cgatctcccc
      661 ggtttcgccg tttgtctcga tcagaggccg cttccggatc tcgccgttgg ccagggtaat
      721 ctcggtcttg aaaaagttca tgatgttgct gtagaagaag tacttggcgg tagccttgcc
      781 gatttcctgc tcgctcttgg cgatcatctt ccgcacgtcg tacaccttgt agtcgccgta
      841 cacgaactcg ctttccagct tagggtactt tttgatcagg gcggttccca cgacggcgtt
      901 caggtaggcg tcgtgggcgt ggtggtagtt gttgatctcg cgcactttgt aaaactggaa
      961 atccttccgg aaatcggaca ccagcttgga cttcagggtg atcactttca cttcccggat
     1021 cagcttgtca ttctcgtcgt acttagtgtt catccgggag tccaggatct gtgccacgtg
     1081 ctttgtgatc tgccgggttt ccaccagctg tctcttgatg aagccggcct tatccagttc
     1141 gctcaggccg cctctctcgg ccttggtcag attgtcgaac tttctctggg taatcagctt
     1201 ggcgttcagc agctgccgcc agtagttctt catcttcttc acgacctctt cggagggcac
     1261 gttgtcgctc ttgccccggt tcttgtcgct tctggtcagc accttgttgt cgatggagtc
     1321 gtccttcaga aagctctgag gcacgatagc gtccacatcg tagtcggaca gccggttgat
     1381 gtccagttcc tggtccacgt acatatcccg cccattctgc aggtagtaca ggtacagctt
     1441 ctcgttctgc agctgggtgt tttccacggg gtgttctttc aggatctggc tgcccagctc
     1501 tttgatgccc tcttcgatcc gcttcattct ctcgcggctg ttcttctgtc ccttctgggt
     1561 ggtctggttc tctctggcca tttcgatcac gatgttctcg ggcttgtgcc ggcccatcac
     1621 tttcacgagc tcgtccacca ccttcactgt ctgcaggatg cccttcttaa tggcggggct
     1681 gccggccaga ttggcaatgt gctcgtgcag gctatcgccc tggccggaca cctgggcttt
     1741 ctggatgtcc tctttaaagg tcaggctgtc gtcgtggatc agctgcatga agtttctgtt
     1801 ggcgaagccg tcggacttca ggaaatccag gattgtcttg ccggactgct tgtcccggat
     1861 gccgttgatc agcttccggc tcagcctgcc ccagccggtg tatctccgcc gcttcagctg
     1921 cttcatcact ttgtcgtcga acaggtgggc ataggttttc agccgttcct cgatcatctc
     1981 tctgtcctca aacagtgtca gggtcagcac gatatcttcc agaatgtcct cgttttcctc
     2041 attgtccagg aagtccttgt ccttgataat tttcagcaga tcgtggtatg tgcccaggga
     2101 ggcgttgaac cgatcttcca cgccggagat ttccacggag tcgaagcact cgattttctt
     2161 gaagtagtcc tctttcagct gcttcacggt cactttccgg ttggtcttga acagcaggtc
     2221 cacgatggcc tttttctgct cgccgctcag gaaggcgggc tttctcattc cctcggtcac
     2281 gtatttcact ttggtcagct cgttatacac ggtgaagtac tcgtacagca ggctgtgctt
     2341 gggcagcacc ttctcgttgg gcaggttctt atcgaagttg gtcatccgct cgatgaagct
     2401 ctgggcggaa gcgcccttgt ccaccacttc ctcgaagttc cagggggtga tggtttcctc
     2461 gctctttctg gtcatccagg cgaatctgct gtttcccctg gccagagggc ccacgtagta
     2521 ggggatgcgg aaggtcagga tcttctcgat cttttcccgg ttgtccttca ggaatgggta
     2581 aaaatcttcc tgccgccgca gaatggcgtg cagctctccc aggtggatct ggtgggggat
     2641 gctgccgttg tcgaaggtcc gctgcttccg cagcaggtcc tctctgttca gcttcacgag
     2701 cagttcctcg gtgccgtcca tcttttccag gatgggcttg atgaacttgt agaactcttc
     2761 ctggctggct ccgccgtcaa tgtagccggc gtagccgttc ttgctctggt cgaagaaaat
     2821 ctctttgtac ttctcaggca gctgctgccg cacgagagct ttcagcaggg tcaggtcctg
     2881 gtggtgctcg tcgtatctct tgatcataga ggcgctcagg ggggccttgg tgatctcggt
     2941 gttcactctc aggatgtcgc tcagcaggat ggcgtcggac aggttcttgg cggccagaaa
     3001 caggtcggcg tactggtcgc cgatctgggc cagcaggttg tccaggtcgt cgtcgtaggt
     3061 gtccttgctc agctgcagtt tggcatcctc ggccaggtcg aagttgctct tgaagttggg
     3121 ggtcaggccc aggctcaggg caatcaggtt gccgaacagg ccattcttct tctcgccggg
     3181 cagctgggcg atcagatttt ccagccgtct gctcttgctc agtctggcag acaggatggc
     3241 cttggcgtcc acgccgctgg cgttgatggg gttttcctcg aacagctggt tgtaggtctg
     3301 caccagctgg atgaacagct tgtccacgtc gctgttgtcg gggttcaggt cgccctcgat
     3361 caggaagtgg ccccggaact tgatcatgtg ggccagggcc agatagatca gccgcaggtc
     3421 ggccttgtcg gtgctgtcca ccagtttctt tctcaggtgg tagatggtgg ggtacttctc
     3481 gtggtaggcc acctcgtcca cgatgttgcc gaagatgggg tgccgctcgt gcttcttatc
     3541 ctcttccacc aggaaggact cttccagtct gtggaagaag ctgtcgtcca ccttggccat
     3601 ctcgttgctg aagatctctt gcagatagca gatccggttc ttccgtctgg tgtatcttct
     3661 tctggcggtt ctcttcagcc gggtggcctc ggctgtttcg ccgctgtcga acagcagggc
     3721 tccgatcagg ttcttcttga tgctgtgccg gtcggtgttg cccagcacct tgaatttctt
     3781 gctgggcacc ttgtactcgt cggtgatcac ggcccagccc acagagttgg tgccgatggc
     3841 caggccgatg ctgtacttct tgtcggctgc tgggactccg tggataccga ccttccgctt
     3901 cttctttggg gccatcttat cgtcatcgtc tttgtaatca atatcatgat ccttgtagtc
     3961 tccgtcgtgg tccttatagt ccatggtggc accggtccaa cctgaaaaaa agtgatttca
     4021 ggcaggtgct ccaggtaatt aaacattaat accccaccaa ccaaccatcc cttaaaccct
     4081 tacctcttgc tcagctaatt acagcccgga ggagaagggc cgtcccgccc gctcacctgt
     4141 gggagtaacg cggtcagtca gagccggggc gggcggcgcg aggcggcggc ggagcggggc
     4201 acggggcgaa ggcagcgcgc agcgactccc gcccgccgcg cgcttcgctt tttatagggc
     4261 cgccgccgcc gccgcctcgc cataaaagga aactttcgga gcgcgccgct ctgattggct
     4321 gccgccgcac ctctccgcct cgccccgccc cgcccctcgc cccgccccgc cccgcctggc
     4381 gcgcgccccc cccccccccc cgcccccatc gctgcacaaa ataattaaaa aataaataaa
     4441 tacaaaattg ggggtgggga ggggggggag atggggagag tgaagcagaa cgtggggctc
     4501 acctcgacca tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc
     4561 cataaggtca tgtactgggc acaatgccag gcgggccatt taccgtcatt gacgtcaata
     4621 gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa
     4681 tactccaccc attgacgtca atggaaagtc cctattggcg ttactattga cgtcaatggg
     4741 cgggggtcgt tgggcggtca gccaggcggg ccatttaccg taagttatgt aacgggtacc
     4801 tctagagcca tttgtctgca gaattggcgc acgcgctaaa aacggactag ccttatttta
     4861 acttgctatt tctagctcta aaacaaaaaa gcaccgactc ggtgccactt tttcaagttg
     4921 ataacggact agccttattt taacttgcta tttctagctc taaaacaggt cttctcgaag
     4981 acccggtgtt tcgtcctttc cacaagatat ataaagccaa gaaatcgaaa tactttcaag
     5041 ttacggtaag catatgatag tccattttaa aacataattt taaaactgca aactacccaa
     5101 gaaattatta ctttctacgt cacgtatttt gtactaatat ctttgtgttt acagtcaaat
     5161 taattccaat tatctctcta acagccttgt atcgtatatg caaatatgaa ggaatcatgg
     5221 gaaataggcc ctcacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     5281 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     5341 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     5401 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     5461 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     5521 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     5581 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     5641 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     5701 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     5761 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     5821 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     5881 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     5941 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     6001 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     6061 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     6121 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     6181 gcaatgatac cgcggcttcc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     6241 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     6301 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     6361 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     6421 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     6481 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     6541 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     6601 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     6661 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     6721 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     6781 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     6841 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     6901 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     6961 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     7021 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     7081 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     7141 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     7201 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     7261 tatgcggcat cagagcagat tgtactgaga gtgcaccata aaattgtaaa cgttaatatt
     7321 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca atagaccgaa
     7381 atcggcaaaa tcccttataa atcaaaagaa tagcccgaga tagagttgag tgttgttcca
     7441 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     7501 gtctatcagg gcgatggccc actacgtgaa ccatcaccca aatcaagttt tttggggtcg
     7561 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     7621 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctaag
     7681 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     7741 ccgctacagg gcgcgtacta tggttgcttt gacgtatgcg gtgtgaaata ccgcacagat
     7801 gcgtaaggag aaaataccgc atcaggcgcc cctgcaggca gctgcgcgct cgctcgctca
     7861 ctgaggccgc ccgggcaaag cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga
     7921 gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc tgcggccgcc
     7981 gcgttaagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     8041 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     8101 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
     8161 gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg ctgattatga tcagttatct
     8221 agatccggtg gatcccgggc ccgcggtacc gtcgactgca gaattcgaag ctttcaggca
     8281 ccgggcttgc gggtcatgca ccaggtgcgc ggtccttcgg gcacctcgac gtcggcggtg
     8341 acggtgaagc cgagccgctc gtagaagggg aggttgtggg gcgcggaggt ctccaggaag
     8401 gcgggcaccc cggcgcgctc ggccgcctcc actccgggga gcacgacggc gctgcccaga
     8461 cccttgccct ggtggtcggg cgagacgccg acggtggcca ggaaccacgc gggctccttg
     8521 ggccggtgcg gcgccaggag gccttccatc tgttgctgcg cggccagccg ggaaccgctc
     8581 aactcggcca tgcgcgggcc gatctcggcg aacaccgccc ccgcttcgac gctctccggc
     8641 gtggtccaga ccgccaccgc ggcgccgtcg tccgcgaccc acaccttgcc gatgtcgagc
     8701 ccgacgcgcg tgaggaagag ttcttgcagc tcggtgaccc gctcgatgtg gcggtccgga
     8761 tcgacggtgt ggcgcgtggc ggggtagtcg gcgaacgcgg cggcgagggt gcgtacggcc
     8821 ctggggacgt cgtcgcgggt ggcgaggcgc accgtgggct tgtactcggt cattgggcca
     8881 ggattctcct cgacgtcacc gcatgttagc agacttcctc tgccctcaga agcttgagct
     8941 cgagatctga gtccggactt gtacagctcg tccatgccga gagtgatccc ggcggcggtc
     9001 acgaactcca gcaggaccat gtgatcgcgc ttctcgttgg ggtctttgct cagggcggac
     9061 tgggtgctca ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat gggggtgttc
     9121 tgctggtagt ggtcggcgag ctgcacgctg ccgtcctcga tgttgtggcg gatcttgaag
     9181 ttcaccttga tgccgttctt ctgcttgtcg gccatgatat agacgttgtg gctgttgtag
     9241 ttgtactcca gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat gcccttcagc
     9301 tcgatgcggt tcaccagggt gtcgccctcg aacttcacct cggcgcgggt cttgtagttg
     9361 ccgtcgtcct tgaagaagat ggtgcgctcc tggacgtagc cttcgggcat ggcggacttg
     9421 aagaagtcgt gctgcttcat gtggtcgggg tagcggctga agcactgcac gccgtaggtc
     9481 agggtggtca cgagggtggg ccagggcacg ggcagcttgc cggtggtgca gatgaacttc
     9541 agggtcagct tgccgtaggt ggcatcgccc tcgccctcgc cggacacgct gaacttgtgg
     9601 ccgtttacgt cgccgtccag ctcgaccagg atgggcacca ccccggtgaa cagctcctcg
     9661 cccttgctca ccatggtggc gaccggtagc gctagcggat ctgacggttc actaaaccag
     9721 ctctgcttat atagacctcc caccgtacac gcctaccgcc catttgcgtc aatggggcgg
     9781 agttgttacg acattttgga aagtcccgtt gattttggtg ccaaaacaaa ctcccattga
     9841 cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca cgcccattga
     9901 tgtactgcca aaaccgcatc accatggtaa tagcgatgac taatacgtag atgtactgcc
     9961 aagtaggaaa gtcccataag gtcatgtact gggcacaatg ccaggcgggc catttaccgt
    10021 cattgacgtc aatagggggc gtacttggca tatgatacac ttgatgtact gccaagtggg
    10081 cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta
    10141 tgggaacata cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg
    10201 ggccatttac cgtaagttat gtaacgcgga acgcggccgc tccccagcat gcctgctatt
    10261 ctcttcccaa tcctccccct tgctgtcctg ccccacccca ccccccagaa tagaatgaca
    10321 cctactcaga caatgcgatg caatttcctc attttattag gaaaggacag tgggagtggc
    10381 accttccagg gtcaaggaag gcacggggga ggggcaaaca acagatggct ggcaactaga
    10441 aggcacagtc gaggctgatc agcgagctct agttagaatt gaattcgggc cggccggaca
    10501 cacacgcaaa atactccttc agcggagcga agaggtggcg gatgactggc acgctccatg
    10561 accggcccag cagtctctgc ctcgccaagc ggctcatgtt ggagacgtca gtatagtgga
    10621 ctgggaaacc aaataccctt tccatttcag tgcaccataa gatgtcctct ttctcattca
    10681 tgaacacagg aaaatgctgg tctttgccct gctttatgga gtttgacctc gtagtaatgg
    10741 tcctcacttt gctgaacttg gctatcctgc catgctccag acactcctgc agctccagct
    10801 tatcattcac agtggatgcc aacggcctgt tcataccggg aaggttaccc cagaagtagc
    10861 gggccctgtg tgcagctgac acttctttgg catcaatcat cacagggttg gactcgagaa
    10921 atcgcgagat gtccctcttg tcactaacgc ccatggccac cacattcgca aagagccaga
    10981 agaaggggcg atcatctccc tccttgggcc gcgcatcatg caggaggcgg tagaactcaa
    11041 agaagagccg gccagtgccc tcgtagaggc ccttgcgagc agggttgacg atggagaggt
    11101 cattgcaggg actgccccca atcaccagat cgaatgggcc ccactcctgg atatgcttct
    11161 gtgtgacgct gcggacgtcc ccgacgtaca tgatcttccc ctggtgccgc accatgccca
    11221 ccgtgatgga gtcctcacac acctccgagg caatgtagcg gtccacctga atgcccaagt
    11281 ccttcagcac caggagccct gtagcgattc catcaaagag agacagcacc cggatgggct
    11341 tcctcttctc agctgggaca ggtgggtaaa cctttggagg gtcaaattcc tggtcgtggt
    11401 tattagcgaa gaacatctgg agccgggagg gggatccgct cccgcctccg ccctcgagct
    11461 ttttcttttt ggcctgtccg gcctttttcg tggccgccgg ccttttgtcg cctcccagct
    11521 gagacaggtc gatccgtgtc tcgtacaggc cggtgatgct ctggtggatc agggtggcgt
    11581 ccagcacctc tttggtgctg gtgtacctct tccggtcgat ggtggtgtca aagtacttga
    11641 aggcggcagg ggctcccaga ttggtcaggg taaacaggtg gatgatattc tcggcctgct
    11701 ctctgatggg cttatcccgg tgcttgttgt aggcggacag cac
//
