LOCUS       sequence-438194-       11979 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses dCAS9-M.SssI 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 11979)
  TITLE     Systemic comparison of dCas9-based DNA methylation editing systems 
            for specificity and stability
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 11979)
  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            "Unpublished"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-M.SssI (dCas9-M.SssI)
FEATURES             Location/Qualifiers
     source          1..11979
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1298..1318)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(1325..1390)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     regulatory      1390..1399
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          1411..1638
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     promoter        complement(1639..1915)
                     /label=chicken beta-actin promoter
     enhancer        1917..2202
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     misc_RNA        complement(2299..2374)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(2393..2641)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(2451..2470)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(2621..2641)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     rep_origin      complement(2703..3291)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2783..2802)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3462..4253,4254..4322))
                     /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     4085..4104
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4323..4427)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4495..4513
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4551..4573)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4673..4692
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(4709..5164)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4846..4867)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5058..5077
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   5239..5379
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5250..5379
                     /label=AAV2 ITR
     polyA_signal    5393..5514
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5404..5423)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5458..5477
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5682..6281)
                     /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     6262..6281
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(6282..6335)
                     /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(6366..7076,7077..7079,7080..7082))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(6366..7076,7077..7079,7080..7082))
                     /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(6408..6429)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     6757..6776
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     regulatory      7079..7088
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(7127..7330)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(7156..7176)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        7331..7634
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    7649..7856
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     7845..7862
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(9104..9151)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(join(9152..10735,10736..10738,10739..11979,
                     1..1246,1247..1249,1250..1273))
                     /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(9152..11979,1..1273))
                     /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 gccgccgcag aatggcgtgc agctctccca ggtggatctg gtgggggatg ctgccgttgt
       61 cgaaggtccg ctgcttccgc agcaggtcct ctctgttcag cttcacgagc agttcctcgg
      121 tgccgtccat cttttccagg atgggcttga tgaacttgta gaactcttcc tggctggctc
      181 cgccgtcaat gtagccggcg tagccgttct tgctctggtc gaagaaaatc tctttgtact
      241 tctcaggcag ctgctgccgc acgagagctt tcagcagggt caggtcctgg tggtgctcgt
      301 cgtatctctt gatcatagag gcgctcaggg gggccttggt gatctcggtg ttcactctca
      361 ggatgtcgct cagcaggatg gcgtcggaca ggttcttggc ggccagaaac aggtcggcgt
      421 actggtcgcc gatctgggcc agcaggttgt ccaggtcgtc gtcgtaggtg tccttgctca
      481 gctgcagttt ggcatcctcg gccaggtcga agttgctctt gaagttgggg gtcaggccca
      541 ggctcagggc aatcaggttg ccgaacaggc cattcttctt ctcgccgggc agctgggcga
      601 tcagattttc cagccgtctg ctcttgctca gtctggcaga caggatggcc ttggcgtcca
      661 cgccgctggc gttgatgggg ttttcctcga acagctggtt gtaggtctgc accagctgga
      721 tgaacagctt gtccacgtcg ctgttgtcgg ggttcaggtc gccctcgatc aggaagtggc
      781 cccggaactt gatcatgtgg gccagggcca gatagatcag ccgcaggtcg gccttgtcgg
      841 tgctgtccac cagtttcttt ctcaggtggt agatggtggg gtacttctcg tggtaggcca
      901 cctcgtccac gatgttgccg aagatggggt gccgctcgtg cttcttatcc tcttccacca
      961 ggaaggactc ttccagtctg tggaagaagc tgtcgtccac cttggccatc tcgttgctga
     1021 agatctcttg cagatagcag atccggttct tccgtctggt gtatcttctt ctggcggttc
     1081 tcttcagccg ggtggcctcg gctgtttcgc cgctgtcgaa cagcagggct ccgatcaggt
     1141 tcttcttgat gctgtgccgg tcggtgttgc ccagcacctt gaatttcttg ctgggcacct
     1201 tgtactcgtc ggtgatcacg gcccagccca cagagttggt gccgatggcc aggccgatgc
     1261 tgtacttctt gtcggctgct gggactccgt ggataccgac cttccgcttc ttctttgggg
     1321 ccatcttatc gtcatcgtct ttgtaatcaa tatcatgatc cttgtagtct ccgtcgtggt
     1381 ccttatagtc catggtggca ccggtccaac ctgaaaaaaa gtgatttcag gcaggtgctc
     1441 caggtaatta aacattaata ccccaccaac caaccatccc ttaaaccctt acctcttgct
     1501 cagctaatta cagcccggag gagaagggcc gtcccgcccg ctcacctgtg ggagtaacgc
     1561 ggtcagtcag agccggggcg ggcggcgcga ggcggcggcg gagcggggca cggggcgaag
     1621 gcagcgcgca gcgactcccg cccgccgcgc gcttcgcttt ttatagggcc gccgccgccg
     1681 ccgcctcgcc ataaaaggaa actttcggag cgcgccgctc tgattggctg ccgccgcacc
     1741 tctccgcctc gccccgcccc gcccctcgcc ccgccccgcc ccgcctggcg cgcgcccccc
     1801 cccccccccg cccccatcgc tgcacaaaat aattaaaaaa taaataaata caaaattggg
     1861 ggtggggagg ggggggagat ggggagagtg aagcagaacg tggggctcac ctcgaccatg
     1921 gtaatagcga tgactaatac gtagatgtac tgccaagtag gaaagtccca taaggtcatg
     1981 tactgggcac aatgccaggc gggccattta ccgtcattga cgtcaatagg gggcgtactt
     2041 ggcatatgat acacttgatg tactgccaag tgggcagttt accgtaaata ctccacccat
     2101 tgacgtcaat ggaaagtccc tattggcgtt actattgacg tcaatgggcg ggggtcgttg
     2161 ggcggtcagc caggcgggcc atttaccgta agttatgtaa cgggtacctc tagagccatt
     2221 tgtctgcaga attggcgcac gcgctaaaaa cggactagcc ttattttaac ttgctatttc
     2281 tagctctaaa acaaaaaagc accgactcgg tgccactttt tcaagttgat aacggactag
     2341 ccttatttta acttgctatt tctagctcta aaacaggtct tctcgaagac ccggtgtttc
     2401 gtcctttcca caagatatat aaagccaaga aatcgaaata ctttcaagtt acggtaagca
     2461 tatgatagtc cattttaaaa cataatttta aaactgcaaa ctacccaaga aattattact
     2521 ttctacgtca cgtattttgt actaatatct ttgtgtttac agtcaaatta attccaatta
     2581 tctctctaac agccttgtat cgtatatgca aatatgaagg aatcatggga aataggccct
     2641 cacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     2701 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     2761 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     2821 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     2881 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     2941 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     3001 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     3061 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     3121 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     3181 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     3241 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     3301 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     3361 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     3421 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     3481 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     3541 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     3601 cggcttccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     3661 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     3721 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     3781 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     3841 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     3901 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     3961 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     4021 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     4081 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     4141 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     4201 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     4261 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     4321 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     4381 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     4441 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     4501 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     4561 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     4621 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     4681 gagcagattg tactgagagt gcaccataaa attgtaaacg ttaatatttt gttaaaattc
     4741 gcgttaaatt tttgttaaat cagctcattt tttaaccaat agaccgaaat cggcaaaatc
     4801 ccttataaat caaaagaata gcccgagata gagttgagtg ttgttccagt ttggaacaag
     4861 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     4921 gatggcccac tacgtgaacc atcacccaaa tcaagttttt tggggtcgag gtgccgtaaa
     4981 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     5041 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctaaggc gctggcaagt
     5101 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     5161 gcgtactatg gttgctttga cgtatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     5221 aataccgcat caggcgcccc tgcaggcagc tgcgcgctcg ctcgctcact gaggccgccc
     5281 gggcaaagcc cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc
     5341 gcagagaggg agtggccaac tccatcacta ggggttcctg cggccgccgc gttaagatac
     5401 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     5461 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     5521 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
     5581 aagtaaaacc tctacaaatg tggtatggct gattatgatc agttatctag atccggtgga
     5641 tcccgggccc gcggtaccgt cgactgcaga attcgaagct ttcaggcacc gggcttgcgg
     5701 gtcatgcacc aggtgcgcgg tccttcgggc acctcgacgt cggcggtgac ggtgaagccg
     5761 agccgctcgt agaaggggag gttgtggggc gcggaggtct ccaggaaggc gggcaccccg
     5821 gcgcgctcgg ccgcctccac tccggggagc acgacggcgc tgcccagacc cttgccctgg
     5881 tggtcgggcg agacgccgac ggtggccagg aaccacgcgg gctccttggg ccggtgcggc
     5941 gccaggaggc cttccatctg ttgctgcgcg gccagccggg aaccgctcaa ctcggccatg
     6001 cgcgggccga tctcggcgaa caccgccccc gcttcgacgc tctccggcgt ggtccagacc
     6061 gccaccgcgg cgccgtcgtc cgcgacccac accttgccga tgtcgagccc gacgcgcgtg
     6121 aggaagagtt cttgcagctc ggtgacccgc tcgatgtggc ggtccggatc gacggtgtgg
     6181 cgcgtggcgg ggtagtcggc gaacgcggcg gcgagggtgc gtacggccct ggggacgtcg
     6241 tcgcgggtgg cgaggcgcac cgtgggcttg tactcggtca ttgggccagg attctcctcg
     6301 acgtcaccgc atgttagcag acttcctctg ccctcagaag cttgagctcg agatctgagt
     6361 ccggacttgt acagctcgtc catgccgaga gtgatcccgg cggcggtcac gaactccagc
     6421 aggaccatgt gatcgcgctt ctcgttgggg tctttgctca gggcggactg ggtgctcagg
     6481 tagtggttgt cgggcagcag cacggggccg tcgccgatgg gggtgttctg ctggtagtgg
     6541 tcggcgagct gcacgctgcc gtcctcgatg ttgtggcgga tcttgaagtt caccttgatg
     6601 ccgttcttct gcttgtcggc catgatatag acgttgtggc tgttgtagtt gtactccagc
     6661 ttgtgcccca ggatgttgcc gtcctccttg aagtcgatgc ccttcagctc gatgcggttc
     6721 accagggtgt cgccctcgaa cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg
     6781 aagaagatgg tgcgctcctg gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc
     6841 tgcttcatgt ggtcggggta gcggctgaag cactgcacgc cgtaggtcag ggtggtcacg
     6901 agggtgggcc agggcacggg cagcttgccg gtggtgcaga tgaacttcag ggtcagcttg
     6961 ccgtaggtgg catcgccctc gccctcgccg gacacgctga acttgtggcc gtttacgtcg
     7021 ccgtccagct cgaccaggat gggcaccacc ccggtgaaca gctcctcgcc cttgctcacc
     7081 atggtggcga ccggtagcgc tagcggatct gacggttcac taaaccagct ctgcttatat
     7141 agacctccca ccgtacacgc ctaccgccca tttgcgtcaa tggggcggag ttgttacgac
     7201 attttggaaa gtcccgttga ttttggtgcc aaaacaaact cccattgacg tcaatggggt
     7261 ggagacttgg aaatccccgt gagtcaaacc gctatccacg cccattgatg tactgccaaa
     7321 accgcatcac catggtaata gcgatgacta atacgtagat gtactgccaa gtaggaaagt
     7381 cccataaggt catgtactgg gcacaatgcc aggcgggcca tttaccgtca ttgacgtcaa
     7441 tagggggcgt acttggcata tgatacactt gatgtactgc caagtgggca gtttaccgta
     7501 aatactccac ccattgacgt caatggaaag tccctattgg cgttactatg ggaacatacg
     7561 tcattattga cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg ccatttaccg
     7621 taagttatgt aacgcggaac gcggccgctc cccagcatgc ctgctattct cttcccaatc
     7681 ctcccccttg ctgtcctgcc ccaccccacc ccccagaata gaatgacacc tactcagaca
     7741 atgcgatgca atttcctcat tttattagga aaggacagtg ggagtggcac cttccagggt
     7801 caaggaaggc acgggggagg ggcaaacaac agatggctgg caactagaag gcacagtcga
     7861 ggctgatcag cgagctctag ttagaattga attcgggccg gccctagcta gggccgccga
     7921 tcttgtcaat gatggcctcc agcacctcca cgctgatgct gttgccacac acgaagatct
     7981 tctggttctc ggtcaggaac tcgatttcgt tcactctctt gccgtcctgg ctgtcaaagc
     8041 cgatgtacag gaaggtctcg tcgctgttca tcttcctgat gttgctgccg tccttgatct
     8101 tgatcctgga gttggcgccg ctagctgtca gtgtggggcc tgtgaactcg gggtcgtaca
     8161 catagccctc ggagttgaac ttggagtagc cgatcaggct ggccttgttg atgttggact
     8221 tggtcttctt gaattcggtg aggttgtatt tcagcaggtt gttgaggatg tccttctcgc
     8281 tcacgatctt gttcagcacc tttttgatgc tcttgggctt cttgtcgccc ttaggcagtt
     8341 ccacgaactc attgagggtg gagatcatga acactctcct cctggcctgg ctggatccga
     8401 aatcggcagc attgaggact tcgatgctgt tctggtagcc caggctctcc agcttctgct
     8461 tccattggtt cagctcctcc tcgttcttct tgtgcagcag ggctcccacg ttctccatga
     8521 ggaggtactt aggcaggtcg ttcttctcgg tggagtccag ggctctttcg atctcccaga
     8581 gcaggccgga tctggtgccg cttcccctct tcatgccctt ctggatgcct agctggctca
     8641 ggtcctggca agggaagctg taggtgagca ggtcgatgtt cttcagtgtc ctcttgtaga
     8701 ggtccctgat gtcgaaaatg ttgccctcct tttcggacag cttgatggcg ttgtagatga
     8761 tcttcagctc gtcatccttc ttcctcttcc agtatccgtt gctgacgggg ttcttgctgt
     8821 tccaggacag ggtcttgttc tcgaggtagt cgatcatctc ctctctgctg acggacttgt
     8881 actcgagctt ggtgtggaag ttgttatgga tggcctggta catgacgatg gcgggcacat
     8941 accactcggc cagtcccacg atctcgtact cgtccttcct cactttctcc agggcctttc
     9001 tctgggcgcc aatgccggcg aaggcttcga acactctcag cttctttgtt ttgttctcca
     9061 ctttgctgtc cctagcggat ccgctcccgc ctccgccctc gagctttttc tttttggcct
     9121 gtccggcctt tttcgtggcc gccggccttt tgtcgcctcc cagctgagac aggtcgatcc
     9181 gtgtctcgta caggccggtg atgctctggt ggatcagggt ggcgtccagc acctctttgg
     9241 tgctggtgta cctcttccgg tcgatggtgg tgtcaaagta cttgaaggcg gcaggggctc
     9301 ccagattggt cagggtaaac aggtggatga tattctcggc ctgctctctg atgggcttat
     9361 cccggtgctt gttgtaggcg gacagcactt tgtccagatt agcgtcggcc aggatcactc
     9421 tcttggagaa ctcgctgatc tgctcgatga tctcgtccag gtagtgcttg tgctgttcca
     9481 caaacagctg tttctgctca ttatcctcgg gggagccctt cagcttctca tagtggctgg
     9541 ccaggtacag gaagttcaca tatttggagg gcagggccag ttcgtttccc ttctgcagtt
     9601 cgccggcaga ggccagcatt ctcttccggc cgttttccag ctcgaacagg gagtacttag
     9661 gcagcttgat gatcaggtcc tttttcactt ctttgtagcc cttggcttcc agaaagtcga
     9721 tgggattctt ctcgaagctg cttctttcca tgatggtgat ccccagcagc tctttcacac
     9781 tcttcagttt cttggacttg cccttttcca ctttggccac caccagcaca gaataggcca
     9841 cggtggggct gtcgaagccg ccgtacttct tagggtccca gtccttcttt ctggcgatca
     9901 gcttatcgct gttcctcttg ggcaggatag actctttgct gaagccgcct gtctgcacct
     9961 cggtcttttt cacgatattc acttggggca tgctcagcac tttccgcacg gtggcaaaat
    10021 cccggccctt atcccacacg atctccccgg tttcgccgtt tgtctcgatc agaggccgct
    10081 tccggatctc gccgttggcc agggtaatct cggtcttgaa aaagttcatg atgttgctgt
    10141 agaagaagta cttggcggta gccttgccga tttcctgctc gctcttggcg atcatcttcc
    10201 gcacgtcgta caccttgtag tcgccgtaca cgaactcgct ttccagctta gggtactttt
    10261 tgatcagggc ggttcccacg acggcgttca ggtaggcgtc gtgggcgtgg tggtagttgt
    10321 tgatctcgcg cactttgtaa aactggaaat ccttccggaa atcggacacc agcttggact
    10381 tcagggtgat cactttcact tcccggatca gcttgtcatt ctcgtcgtac ttagtgttca
    10441 tccgggagtc caggatctgt gccacgtgct ttgtgatctg ccgggtttcc accagctgtc
    10501 tcttgatgaa gccggcctta tccagttcgc tcaggccgcc tctctcggcc ttggtcagat
    10561 tgtcgaactt tctctgggta atcagcttgg cgttcagcag ctgccgccag tagttcttca
    10621 tcttcttcac gacctcttcg gagggcacgt tgtcgctctt gccccggttc ttgtcgcttc
    10681 tggtcagcac cttgttgtcg atggagtcgt ccttcagaaa gctctgaggc acgatagcgt
    10741 ccacatcgta gtcggacagc cggttgatgt ccagttcctg gtccacgtac atatcccgcc
    10801 cattctgcag gtagtacagg tacagcttct cgttctgcag ctgggtgttt tccacggggt
    10861 gttctttcag gatctggctg cccagctctt tgatgccctc ttcgatccgc ttcattctct
    10921 cgcggctgtt cttctgtccc ttctgggtgg tctggttctc tctggccatt tcgatcacga
    10981 tgttctcggg cttgtgccgg cccatcactt tcacgagctc gtccaccacc ttcactgtct
    11041 gcaggatgcc cttcttaatg gcggggctgc cggccagatt ggcaatgtgc tcgtgcaggc
    11101 tatcgccctg gccggacacc tgggctttct ggatgtcctc tttaaaggtc aggctgtcgt
    11161 cgtggatcag ctgcatgaag tttctgttgg cgaagccgtc ggacttcagg aaatccagga
    11221 ttgtcttgcc ggactgcttg tcccggatgc cgttgatcag cttccggctc agcctgcccc
    11281 agccggtgta tctccgccgc ttcagctgct tcatcacttt gtcgtcgaac aggtgggcat
    11341 aggttttcag ccgttcctcg atcatctctc tgtcctcaaa cagtgtcagg gtcagcacga
    11401 tatcttccag aatgtcctcg ttttcctcat tgtccaggaa gtccttgtcc ttgataattt
    11461 tcagcagatc gtggtatgtg cccagggagg cgttgaaccg atcttccacg ccggagattt
    11521 ccacggagtc gaagcactcg attttcttga agtagtcctc tttcagctgc ttcacggtca
    11581 ctttccggtt ggtcttgaac agcaggtcca cgatggcctt tttctgctcg ccgctcagga
    11641 aggcgggctt tctcattccc tcggtcacgt atttcacttt ggtcagctcg ttatacacgg
    11701 tgaagtactc gtacagcagg ctgtgcttgg gcagcacctt ctcgttgggc aggttcttat
    11761 cgaagttggt catccgctcg atgaagctct gggcggaagc gcccttgtcc accacttcct
    11821 cgaagttcca gggggtgatg gtttcctcgc tctttctggt catccaggcg aatctgctgt
    11881 ttcccctggc cagagggccc acgtagtagg ggatgcggaa ggtcaggatc ttctcgatct
    11941 tttcccggtt gtccttcagg aatgggtaaa aatcttcct
//
