LOCUS       sequence-438178-       12762 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses dCAS9-DNMT3A-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 12762)
  TITLE     Systemic comparison of dCas9-based DNA methylation editing systems 
            for specificity and stability
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 12762)
  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            "Unpublished"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-DNMT3A (dCas9-3A-3A)
FEATURES             Location/Qualifiers
     source          1..12762
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    202..409
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     398..415
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(460..1392)
                     /codon_start=1
                     /product="catalytic domain of DNA methyltransferase 3A"
                     /label=DNMT3A catalytic domain
                     /translation="PSRLQMFFANNHDQEFDPPKVYPPVPAEKRKPIRVLSLFDGIATG
                     LLVLKDLGIQVDRYIASEVCEDSITVGMVRHQGKIMYVGDVRSVTQKHIQEWGPFDLVI
                     GGSPCNDLSIVNPARKGLYEGTGRLFFEFYRLLHDARPKEGDDRPFFWLFENVVAMGVS
                     DKRDISRFLESNPVMIDAKEVSAAHRARYFWGNLPGMNRPLASTVNDKLELQECLEHGR
                     IAKFSKVRTITTRSNSIKQGKDQHFPVFMNEKEDILWCTEMERVFGFPVHYTDVSNMSR
                     LARQRLLGRSWSVPVIRHLFAPLKEYFACV"
     CDS             complement(1420..1467)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(1480..2412)
                     /codon_start=1
                     /product="catalytic domain of DNA methyltransferase 3A"
                     /label=DNMT3A catalytic domain
                     /translation="PSRLQMFFANNHDQEFDPPKVYPPVPAEKRKPIRVLSLFDGIATG
                     LLVLKDLGIQVDRYIASEVCEDSITVGMVRHQGKIMYVGDVRSVTQKHIQEWGPFDLVI
                     GGSPCNDLSIVNPARKGLYEGTGRLFFEFYRLLHDARPKEGDDRPFFWLFENVVAMGVS
                     DKRDISRFLESNPVMIDAKEVSAAHRARYFWGNLPGMNRPLASTVNDKLELQECLEHGR
                     IAKFSKVRTITTRSNSIKQGKDQHFPVFMNEKEDILWCTEMERVFGFPVHYTDVSNMSR
                     LARQRLLGRSWSVPVIRHLFAPLKEYFACV"
     CDS             complement(2440..2487)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(join(2488..4071,4072..4074,4075..6561,6562..6564,
                     6565..6588))
                     /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(2488..6588)
                     /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(6613..6633)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(6640..6705)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     regulatory      6705..6714
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          6726..6953
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     promoter        complement(6954..7230)
                     /label=chicken beta-actin promoter
     enhancer        7232..7517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     misc_RNA        complement(7614..7689)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(7708..7956)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(7766..7785)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(7936..7956)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     rep_origin      complement(8018..8606)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8098..8117)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8777..9568,9569..9637))
                     /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     9400..9419
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9638..9742)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9810..9828
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(9866..9888)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     9988..10007
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(10024..10479)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10161..10182)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     10373..10392
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   10554..10694
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   10565..10694
                     /label=AAV2 ITR
     polyA_signal    10708..10829
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10719..10738)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     10773..10792
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(10997..11596)
                     /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     11577..11596
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(11597..11650)
                     /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(11681..12391,12392..12394,12395..12397))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(11681..12391,12392..12394,12395..12397))
                     /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(11723..11744)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     12072..12091
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     regulatory      12394..12403
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(12442..12645)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(12471..12491)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        join(12646..12762,1..187)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 cgtacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc gtaaatactc
       61 cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat acgtcattat
      121 tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc gggccattta ccgtaagtta
      181 tgtaacgcgg aacgcggccg ctccccagca tgcctgctat tctcttccca atcctccccc
      241 ttgctgtcct gccccacccc accccccaga atagaatgac acctactcag acaatgcgat
      301 gcaatttcct cattttatta ggaaaggaca gtgggagtgg caccttccag ggtcaaggaa
      361 ggcacggggg aggggcaaac aacagatggc tggcaactag aaggcacagt cgaggctgat
      421 cagcgagctc tagttagaat tgaattcggg ccggccggac acacacgcaa aatactcctt
      481 cagcggagcg aagaggtggc ggatgactgg cacgctccat gaccggccca gcagtctctg
      541 cctcgccaag cggctcatgt tggagacgtc agtatagtgg actgggaaac caaataccct
      601 ttccatttca gtgcaccata agatgtcctc tttctcattc atgaacacag gaaaatgctg
      661 gtctttgccc tgctttatgg agtttgacct cgtagtaatg gtcctcactt tgctgaactt
      721 ggctatcctg ccatgctcca gacactcctg cagctccagc ttatcattca cagtggatgc
      781 caacggcctg ttcataccgg gaaggttacc ccagaagtag cgggccctgt gtgcagctga
      841 cacttctttg gcatcaatca tcacagggtt ggactcgaga aatcgcgaga tgtccctctt
      901 gtcactaacg cccatggcca ccacattctc aaagagccag aagaaggggc gatcatctcc
      961 ctccttgggc cgcgcatcat gcaggaggcg gtagaactca aagaagagcc ggccagtgcc
     1021 ctcgtagagg cccttgcgag cagggttgac gatggagagg tcattgcagg gactgccccc
     1081 aatcaccaga tcgaatgggc cccactcctg gatatgcttc tgtgtgacgc tgcggacgtc
     1141 cccgacgtac atgatcttcc cctggtgccg caccatgccc accgtgatgg agtcctcaca
     1201 cacctccgag gcaatgtagc ggtccacctg aatgcccaag tccttcagca ccaggagccc
     1261 tgtagcgatt ccatcaaaga gagacagcac ccggatgggc ttcctcttct cagctgggac
     1321 aggtgggtaa acctttggag ggtcaaattc ctggtcgtgg ttattagcga agaacatctg
     1381 gagccgggag ggggatccgc tcccgcctcc gccctcgagc tttttctttt tggcctgtcc
     1441 ggcctttttc gtggccgccg gccttttagg ccggccggac acacacgcaa aatactcctt
     1501 cagcggagcg aagaggtggc ggatgactgg cacgctccat gaccggccca gcagtctctg
     1561 cctcgccaag cggctcatgt tggagacgtc agtatagtgg actgggaaac caaataccct
     1621 ttccatttca gtgcaccata agatgtcctc tttctcattc atgaacacag gaaaatgctg
     1681 gtctttgccc tgctttatgg agtttgacct cgtagtaatg gtcctcactt tgctgaactt
     1741 ggctatcctg ccatgctcca gacactcctg cagctccagc ttatcattca cagtggatgc
     1801 caacggcctg ttcataccgg gaaggttacc ccagaagtag cgggccctgt gtgcagctga
     1861 cacttctttg gcatcaatca tcacagggtt ggactcgaga aatcgcgaga tgtccctctt
     1921 gtcactaacg cccatggcca ccacattctc aaagagccag aagaaggggc gatcatctcc
     1981 ctccttgggc cgcgcatcat gcaggaggcg gtagaactca aagaagagcc ggccagtgcc
     2041 ctcgtagagg cccttgcgag cagggttgac gatggagagg tcattgcagg gactgccccc
     2101 aatcaccaga tcgaatgggc cccactcctg gatatgcttc tgtgtgacgc tgcggacgtc
     2161 cccgacgtac atgatcttcc cctggtgccg caccatgccc accgtgatgg agtcctcaca
     2221 cacctccgag gcaatgtagc ggtccacctg aatgcccaag tccttcagca ccaggagccc
     2281 tgtagcgatt ccatcaaaga gagacagcac ccggatgggc ttcctcttct cagctgggac
     2341 aggtgggtaa acctttggag ggtcaaattc ctggtcgtgg ttattagcga agaacatctg
     2401 gagccgggag ggggatccgc tcccgcctcc gccctcgagc tttttctttt tggcctgtcc
     2461 ggcctttttc gtggccgccg gccttttgtc gcctcccagc tgagacaggt cgatccgtgt
     2521 ctcgtacagg ccggtgatgc tctggtggat cagggtggcg tccagcacct ctttggtgct
     2581 ggtgtacctc ttccggtcga tggtggtgtc aaagtacttg aaggcggcag gggctcccag
     2641 attggtcagg gtaaacaggt ggatgatatt ctcggcctgc tctctgatgg gcttatcccg
     2701 gtgcttgttg taggcggaca gcactttgtc cagattagcg tcggccagga tcactctctt
     2761 ggagaactcg ctgatctgct cgatgatctc gtccaggtag tgcttgtgct gttccacaaa
     2821 cagctgtttc tgctcattat cctcggggga gcccttcagc ttctcatagt ggctggccag
     2881 gtacaggaag ttcacatatt tggagggcag ggccagttcg tttcccttct gcagttcgcc
     2941 ggcagaggcc agcattctct tccggccgtt ttccagctcg aacagggagt acttaggcag
     3001 cttgatgatc aggtcctttt tcacttcttt gtagcccttg gcttccagaa agtcgatggg
     3061 attcttctcg aagctgcttc tttccatgat ggtgatcccc agcagctctt tcacactctt
     3121 cagtttcttg gacttgccct tttccacttt ggccaccacc agcacagaat aggccacggt
     3181 ggggctgtcg aagccgccgt acttcttagg gtcccagtcc ttctttctgg cgatcagctt
     3241 atcgctgttc ctcttgggca ggatagactc tttgctgaag ccgcctgtct gcacctcggt
     3301 ctttttcacg atattcactt ggggcatgct cagcactttc cgcacggtgg caaaatcccg
     3361 gcccttatcc cacacgatct ccccggtttc gccgtttgtc tcgatcagag gccgcttccg
     3421 gatctcgccg ttggccaggg taatctcggt cttgaaaaag ttcatgatgt tgctgtagaa
     3481 gaagtacttg gcggtagcct tgccgatttc ctgctcgctc ttggcgatca tcttccgcac
     3541 gtcgtacacc ttgtagtcgc cgtacacgaa ctcgctttcc agcttagggt actttttgat
     3601 cagggcggtt cccacgacgg cgttcaggta ggcgtcgtgg gcgtggtggt agttgttgat
     3661 ctcgcgcact ttgtaaaact ggaaatcctt ccggaaatcg gacaccagct tggacttcag
     3721 ggtgatcact ttcacttccc ggatcagctt gtcattctcg tcgtacttag tgttcatccg
     3781 ggagtccagg atctgtgcca cgtgctttgt gatctgccgg gtttccacca gctgtctctt
     3841 gatgaagccg gccttatcca gttcgctcag gccgcctctc tcggccttgg tcagattgtc
     3901 gaactttctc tgggtaatca gcttggcgtt cagcagctgc cgccagtagt tcttcatctt
     3961 cttcacgacc tcttcggagg gcacgttgtc gctcttgccc cggttcttgt cgcttctggt
     4021 cagcaccttg ttgtcgatgg agtcgtcctt cagaaagctc tgaggcacga tagcgtccac
     4081 atcgtagtcg gacagccggt tgatgtccag ttcctggtcc acgtacatat cccgcccatt
     4141 ctgcaggtag tacaggtaca gcttctcgtt ctgcagctgg gtgttttcca cggggtgttc
     4201 tttcaggatc tggctgccca gctctttgat gccctcttcg atccgcttca ttctctcgcg
     4261 gctgttcttc tgtcccttct gggtggtctg gttctctctg gccatttcga tcacgatgtt
     4321 ctcgggcttg tgccggccca tcactttcac gagctcgtcc accaccttca ctgtctgcag
     4381 gatgcccttc ttaatggcgg ggctgccggc cagattggca atgtgctcgt gcaggctatc
     4441 gccctggccg gacacctggg ctttctggat gtcctcttta aaggtcaggc tgtcgtcgtg
     4501 gatcagctgc atgaagtttc tgttggcgaa gccgtcggac ttcaggaaat ccaggattgt
     4561 cttgccggac tgcttgtccc ggatgccgtt gatcagcttc cggctcagcc tgccccagcc
     4621 ggtgtatctc cgccgcttca gctgcttcat cactttgtcg tcgaacaggt gggcataggt
     4681 tttcagccgt tcctcgatca tctctctgtc ctcaaacagt gtcagggtca gcacgatatc
     4741 ttccagaatg tcctcgtttt cctcattgtc caggaagtcc ttgtccttga taattttcag
     4801 cagatcgtgg tatgtgccca gggaggcgtt gaaccgatct tccacgccgg agatttccac
     4861 ggagtcgaag cactcgattt tcttgaagta gtcctctttc agctgcttca cggtcacttt
     4921 ccggttggtc ttgaacagca ggtccacgat ggcctttttc tgctcgccgc tcaggaaggc
     4981 gggctttctc attccctcgg tcacgtattt cactttggtc agctcgttat acacggtgaa
     5041 gtactcgtac agcaggctgt gcttgggcag caccttctcg ttgggcaggt tcttatcgaa
     5101 gttggtcatc cgctcgatga agctctgggc ggaagcgccc ttgtccacca cttcctcgaa
     5161 gttccagggg gtgatggttt cctcgctctt tctggtcatc caggcgaatc tgctgtttcc
     5221 cctggccaga gggcccacgt agtaggggat gcggaaggtc aggatcttct cgatcttttc
     5281 ccggttgtcc ttcaggaatg ggtaaaaatc ttcctgccgc cgcagaatgg cgtgcagctc
     5341 tcccaggtgg atctggtggg ggatgctgcc gttgtcgaag gtccgctgct tccgcagcag
     5401 gtcctctctg ttcagcttca cgagcagttc ctcggtgccg tccatctttt ccaggatggg
     5461 cttgatgaac ttgtagaact cttcctggct ggctccgccg tcaatgtagc cggcgtagcc
     5521 gttcttgctc tggtcgaaga aaatctcttt gtacttctca ggcagctgct gccgcacgag
     5581 agctttcagc agggtcaggt cctggtggtg ctcgtcgtat ctcttgatca tagaggcgct
     5641 caggggggcc ttggtgatct cggtgttcac tctcaggatg tcgctcagca ggatggcgtc
     5701 ggacaggttc ttggcggcca gaaacaggtc ggcgtactgg tcgccgatct gggccagcag
     5761 gttgtccagg tcgtcgtcgt aggtgtcctt gctcagctgc agtttggcat cctcggccag
     5821 gtcgaagttg ctcttgaagt tgggggtcag gcccaggctc agggcaatca ggttgccgaa
     5881 caggccattc ttcttctcgc cgggcagctg ggcgatcaga ttttccagcc gtctgctctt
     5941 gctcagtctg gcagacagga tggccttggc gtccacgccg ctggcgttga tggggttttc
     6001 ctcgaacagc tggttgtagg tctgcaccag ctggatgaac agcttgtcca cgtcgctgtt
     6061 gtcggggttc aggtcgccct cgatcaggaa gtggccccgg aacttgatca tgtgggccag
     6121 ggccagatag atcagccgca ggtcggcctt gtcggtgctg tccaccagtt tctttctcag
     6181 gtggtagatg gtggggtact tctcgtggta ggccacctcg tccacgatgt tgccgaagat
     6241 ggggtgccgc tcgtgcttct tatcctcttc caccaggaag gactcttcca gtctgtggaa
     6301 gaagctgtcg tccaccttgg ccatctcgtt gctgaagatc tcttgcagat agcagatccg
     6361 gttcttccgt ctggtgtatc ttcttctggc ggttctcttc agccgggtgg cctcggctgt
     6421 ttcgccgctg tcgaacagca gggctccgat caggttcttc ttgatgctgt gccggtcggt
     6481 gttgcccagc accttgaatt tcttgctggg caccttgtac tcgtcggtga tcacggccca
     6541 gcccacagag ttggtgccga tggccaggcc gatgctgtac ttcttgtcgg ctgctgggac
     6601 tccgtggata ccgaccttcc gcttcttctt tggggccatc ttatcgtcat cgtctttgta
     6661 atcaatatca tgatccttgt agtctccgtc gtggtcctta tagtccatgg tggcaccggt
     6721 ccaacctgaa aaaaagtgat ttcaggcagg tgctccaggt aattaaacat taatacccca
     6781 ccaaccaacc atcccttaaa cccttacctc ttgctcagct aattacagcc cggaggagaa
     6841 gggccgtccc gcccgctcac ctgtgggagt aacgcggtca gtcagagccg gggcgggcgg
     6901 cgcgaggcgg cggcggagcg gggcacgggg cgaaggcagc gcgcagcgac tcccgcccgc
     6961 cgcgcgcttc gctttttata gggccgccgc cgccgccgcc tcgccataaa aggaaacttt
     7021 cggagcgcgc cgctctgatt ggctgccgcc gcacctctcc gcctcgcccc gccccgcccc
     7081 tcgccccgcc ccgccccgcc tggcgcgcgc cccccccccc ccccgccccc atcgctgcac
     7141 aaaataatta aaaaataaat aaatacaaaa ttgggggtgg ggaggggggg gagatgggga
     7201 gagtgaagca gaacgtgggg ctcacctcga ccatggtaat agcgatgact aatacgtaga
     7261 tgtactgcca agtaggaaag tcccataagg tcatgtactg ggcacaatgc caggcgggcc
     7321 atttaccgtc attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg
     7381 ccaagtgggc agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg
     7441 gcgttactat tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc gggccattta
     7501 ccgtaagtta tgtaacgggt acctctagag ccatttgtct gcagaattgg cgcacgcgct
     7561 aaaaacggac tagccttatt ttaacttgct atttctagct ctaaaacaaa aaagcaccga
     7621 ctcggtgcca ctttttcaag ttgataacgg actagcctta ttttaacttg ctatttctag
     7681 ctctaaaaca ggtcttctcg aagacccggt gtttcgtcct ttccacaaga tatataaagc
     7741 caagaaatcg aaatactttc aagttacggt aagcatatga tagtccattt taaaacataa
     7801 ttttaaaact gcaaactacc caagaaatta ttactttcta cgtcacgtat tttgtactaa
     7861 tatctttgtg tttacagtca aattaattcc aattatctct ctaacagcct tgtatcgtat
     7921 atgcaaatat gaaggaatca tgggaaatag gccctcacat gtgagcaaaa ggccagcaaa
     7981 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     8041 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     8101 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     8161 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     8221 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     8281 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     8341 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     8401 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     8461 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     8521 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     8581 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     8641 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     8701 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     8761 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     8821 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     8881 gcttaccatc tggccccagt gctgcaatga taccgcggct tccacgctca ccggctccag
     8941 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     9001 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     9061 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     9121 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     9181 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     9241 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     9301 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     9361 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     9421 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     9481 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     9541 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     9601 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     9661 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     9721 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     9781 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     9841 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     9901 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     9961 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
    10021 ataaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
    10081 cattttttaa ccaatagacc gaaatcggca aaatccctta taaatcaaaa gaatagcccg
    10141 agatagagtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
    10201 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
    10261 ccaaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
    10321 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
    10381 aagcgaaagg agcgggcgct aaggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
    10441 ccacacccgc cgcgcttaat gcgccgctac agggcgcgta ctatggttgc tttgacgtat
    10501 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gcccctgcag
    10561 gcagctgcgc gctcgctcgc tcactgaggc cgcccgggca aagcccgggc gtcgggcgac
    10621 ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat
    10681 cactaggggt tcctgcggcc gccgcgttaa gatacattga tgagtttgga caaaccacaa
    10741 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
    10801 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
    10861 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
    10921 tggctgatta tgatcagtta tctagatccg gtggatcccg ggcccgcggt accgtcgact
    10981 gcagaattcg aagctttcag gcaccgggct tgcgggtcat gcaccaggtg cgcggtcctt
    11041 cgggcacctc gacgtcggcg gtgacggtga agccgagccg ctcgtagaag gggaggttgt
    11101 ggggcgcgga ggtctccagg aaggcgggca ccccggcgcg ctcggccgcc tccactccgg
    11161 ggagcacgac ggcgctgccc agacccttgc cctggtggtc gggcgagacg ccgacggtgg
    11221 ccaggaacca cgcgggctcc ttgggccggt gcggcgccag gaggccttcc atctgttgct
    11281 gcgcggccag ccgggaaccg ctcaactcgg ccatgcgcgg gccgatctcg gcgaacaccg
    11341 cccccgcttc gacgctctcc ggcgtggtcc agaccgccac cgcggcgccg tcgtccgcga
    11401 cccacacctt gccgatgtcg agcccgacgc gcgtgaggaa gagttcttgc agctcggtga
    11461 cccgctcgat gtggcggtcc ggatcgacgg tgtggcgcgt ggcggggtag tcggcgaacg
    11521 cggcggcgag ggtgcgtacg gccctgggga cgtcgtcgcg ggtggcgagg cgcaccgtgg
    11581 gcttgtactc ggtcattggg ccaggattct cctcgacgtc accgcatgtt agcagacttc
    11641 ctctgccctc agaagcttga gctcgagatc tgagtccgga cttgtacagc tcgtccatgc
    11701 cgagagtgat cccggcggcg gtcacgaact ccagcaggac catgtgatcg cgcttctcgt
    11761 tggggtcttt gctcagggcg gactgggtgc tcaggtagtg gttgtcgggc agcagcacgg
    11821 ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc gagctgcacg ctgccgtcct
    11881 cgatgttgtg gcggatcttg aagttcacct tgatgccgtt cttctgcttg tcggccatga
    11941 tatagacgtt gtggctgttg tagttgtact ccagcttgtg ccccaggatg ttgccgtcct
    12001 ccttgaagtc gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca
    12061 cctcggcgcg ggtcttgtag ttgccgtcgt ccttgaagaa gatggtgcgc tcctggacgt
    12121 agccttcggg catggcggac ttgaagaagt cgtgctgctt catgtggtcg gggtagcggc
    12181 tgaagcactg cacgccgtag gtcagggtgg tcacgagggt gggccagggc acgggcagct
    12241 tgccggtggt gcagatgaac ttcagggtca gcttgccgta ggtggcatcg ccctcgccct
    12301 cgccggacac gctgaacttg tggccgttta cgtcgccgtc cagctcgacc aggatgggca
    12361 ccaccccggt gaacagctcc tcgcccttgc tcaccatggt ggcgaccggt agcgctagcg
    12421 gatctgacgg ttcactaaac cagctctgct tatatagacc tcccaccgta cacgcctacc
    12481 gcccatttgc gtcaatgggg cggagttgtt acgacatttt ggaaagtccc gttgattttg
    12541 gtgccaaaac aaactcccat tgacgtcaat ggggtggaga cttggaaatc cccgtgagtc
    12601 aaaccgctat ccacgcccat tgatgtactg ccaaaaccgc atcaccatgg taatagcgat
    12661 gactaatacg tagatgtact gccaagtagg aaagtcccat aaggtcatgt actgggcaca
    12721 atgccaggcg ggccatttac cgtcattgac gtcaataggg gg
//
