LOCUS       sequence-438174-       11741 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses dCAS9-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 11741)
  TITLE     Systemic comparison of dCas9-based DNA methylation editing systems 
            for specificity and stability
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 11741)
  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            "Unpublished"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
FEATURES             Location/Qualifiers
     source          1..11741
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             510..557
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             585..1517
                     /codon_start=1
                     /product="catalytic domain of DNA methyltransferase 3A"
                     /label=DNMT3A catalytic domain
                     /translation="PSRLQMFFANNHDQEFDPPKVYPPVPAEKRKPIRVLSLFDGIATG
                     LLVLKDLGIQVDRYIASEVCEDSITVGMVRHQGKIMYVGDVRSVTQKHIQEWGPFDLVI
                     GGSPCNDLSIVNPARKGLYEGTGRLFFEFYRLLHDARPKEGDDRPFFWLFENVVAMGVS
                     DKRDISRFLESNPVMIDAKEVSAAHRARYFWGNLPGMNRPLASTVNDKLELQECLEHGR
                     IAKFSKVRTITTRSNSIKQGKDQHFPVFMNEKEDILWCTEMERVFGFPVHYTDVSNMSR
                     LARQRLLGRSWSVPVIRHLFAPLKEYFACV"
     primer_bind     complement(1562..1579)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1568..1775
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    1789..1910
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1800..1819)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1854..1873
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2078..2677)
                     /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     2658..2677
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(2678..2731)
                     /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(2762..3472,3473..3475,3476..3478))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(2762..3472,3473..3475,3476..3478))
                     /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(2804..2825)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3153..3172
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     regulatory      3475..3484
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3523..3726)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3552..3572)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3727..4030
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     repeat_region   4045..4185
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4045..4174
                     /label=AAV2 ITR
     rep_origin      4260..4715
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4347..4366)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4557..4578
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4732..4751)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4851..4873
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4911..4929)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4997..5101
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5102..5170,5171..5962)
                     /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     complement(5320..5339)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6133..6721
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6622..6641
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        6783..7031
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6783..6803
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6954..6973
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        7050..7125
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        7222..7507
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        7509..7784
                     /label=chicken beta-actin promoter
     intron          7785..8012
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      8024..8033
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             8033..8098
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             8105..8125
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(8150..8173,8174..8176,8177..10663,10664..10666,
                     10667..11741,1..509)
                     /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             join(8150..11741,1..509)
                     /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 ctaagtactc cctgttcgag ctggaaaacg gccggaagag aatgctggcc tctgccggcg
       61 aactgcagaa gggaaacgaa ctggccctgc cctccaaata tgtgaacttc ctgtacctgg
      121 ccagccacta tgagaagctg aagggctccc ccgaggataa tgagcagaaa cagctgtttg
      181 tggaacagca caagcactac ctggacgaga tcatcgagca gatcagcgag ttctccaaga
      241 gagtgatcct ggccgacgct aatctggaca aagtgctgtc cgcctacaac aagcaccggg
      301 ataagcccat cagagagcag gccgagaata tcatccacct gtttaccctg accaatctgg
      361 gagcccctgc cgccttcaag tactttgaca ccaccatcga ccggaagagg tacaccagca
      421 ccaaagaggt gctggacgcc accctgatcc accagagcat caccggcctg tacgagacac
      481 ggatcgacct gtctcagctg ggaggcgaca aaaggccggc ggccacgaaa aaggccggac
      541 aggccaaaaa gaaaaagctc gagggcggag gcgggagcgg atccccctcc cggctccaga
      601 tgttcttcgc taataaccac gaccaggaat ttgaccctcc aaaggtttac ccacctgtcc
      661 cagctgagaa gaggaagccc atccgggtgc tgtctctctt tgatggaatc gctacagggc
      721 tcctggtgct gaaggacttg ggcattcagg tggaccgcta cattgcctcg gaggtgtgtg
      781 aggactccat cacggtgggc atggtgcggc accaggggaa gatcatgtac gtcggggacg
      841 tccgcagcgt cacacagaag catatccagg agtggggccc attcgatctg gtgattgggg
      901 gcagtccctg caatgacctc tccatcgtca accctgctcg caagggcctc tacgagggca
      961 ctggccggct cttctttgag ttctaccgcc tcctgcatga tgcgcggccc aaggagggag
     1021 atgatcgccc cttcttctgg ctctttgaga atgtggtggc catgggcgtt agtgacaaga
     1081 gggacatctc gcgatttctc gagtccaacc ctgtgatgat tgatgccaaa gaagtgtcag
     1141 ctgcacacag ggcccgctac ttctggggta accttcccgg tatgaacagg ccgttggcat
     1201 ccactgtgaa tgataagctg gagctgcagg agtgtctgga gcatggcagg atagccaagt
     1261 tcagcaaagt gaggaccatt actacgaggt caaactccat aaagcagggc aaagaccagc
     1321 attttcctgt gttcatgaat gagaaagagg acatcttatg gtgcactgaa atggaaaggg
     1381 tatttggttt cccagtccac tatactgacg tctccaacat gagccgcttg gcgaggcaga
     1441 gactgctggg ccggtcatgg agcgtgccag tcatccgcca cctcttcgct ccgctgaagg
     1501 agtattttgc gtgtgtgtcc ggccggcccg aattcaattc taactagagc tcgctgatca
     1561 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     1621 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     1681 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     1741 gaggattggg aagagaatag caggcatgct ggggagcggc cgccgcgtta agatacattg
     1801 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     1861 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     1921 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
     1981 aaaacctcta caaatgtggt atggctgatt atgatcagtt atctagatcc ggtggatccc
     2041 gggcccgcgg taccgtcgac tgcagaattc gaagctttca ggcaccgggc ttgcgggtca
     2101 tgcaccaggt gcgcggtcct tcgggcacct cgacgtcggc ggtgacggtg aagccgagcc
     2161 gctcgtagaa ggggaggttg tggggcgcgg aggtctccag gaaggcgggc accccggcgc
     2221 gctcggccgc ctccactccg gggagcacga cggcgctgcc cagacccttg ccctggtggt
     2281 cgggcgagac gccgacggtg gccaggaacc acgcgggctc cttgggccgg tgcggcgcca
     2341 ggaggccttc catctgttgc tgcgcggcca gccgggaacc gctcaactcg gccatgcgcg
     2401 ggccgatctc ggcgaacacc gcccccgctt cgacgctctc cggcgtggtc cagaccgcca
     2461 ccgcggcgcc gtcgtccgcg acccacacct tgccgatgtc gagcccgacg cgcgtgagga
     2521 agagttcttg cagctcggtg acccgctcga tgtggcggtc cggatcgacg gtgtggcgcg
     2581 tggcggggta gtcggcgaac gcggcggcga gggtgcgtac ggccctgggg acgtcgtcgc
     2641 gggtggcgag gcgcaccgtg ggcttgtact cggtcattgg gccaggattc tcctcgacgt
     2701 caccgcatgt tagcagactt cctctgccct cagaagcttg agctcgagat ctgagtccgg
     2761 acttgtacag ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac tccagcagga
     2821 ccatgtgatc gcgcttctcg ttggggtctt tgctcagggc ggactgggtg ctcaggtagt
     2881 ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg
     2941 cgagctgcac gctgccgtcc tcgatgttgt ggcggatctt gaagttcacc ttgatgccgt
     3001 tcttctgctt gtcggccatg atatagacgt tgtggctgtt gtagttgtac tccagcttgt
     3061 gccccaggat gttgccgtcc tccttgaagt cgatgccctt cagctcgatg cggttcacca
     3121 gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg tccttgaaga
     3181 agatggtgcg ctcctggacg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct
     3241 tcatgtggtc ggggtagcgg ctgaagcact gcacgccgta ggtcagggtg gtcacgaggg
     3301 tgggccaggg cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt
     3361 aggtggcatc gccctcgccc tcgccggaca cgctgaactt gtggccgttt acgtcgccgt
     3421 ccagctcgac caggatgggc accaccccgg tgaacagctc ctcgcccttg ctcaccatgg
     3481 tggcgaccgg tagcgctagc ggatctgacg gttcactaaa ccagctctgc ttatatagac
     3541 ctcccaccgt acacgcctac cgcccatttg cgtcaatggg gcggagttgt tacgacattt
     3601 tggaaagtcc cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa tggggtggag
     3661 acttggaaat ccccgtgagt caaaccgcta tccacgccca ttgatgtact gccaaaaccg
     3721 catcaccatg gtaatagcga tgactaatac gtagatgtac tgccaagtag gaaagtccca
     3781 taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga cgtcaatagg
     3841 gggcgtactt ggcatatgat acacttgatg tactgccaag tgggcagttt accgtaaata
     3901 ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa catacgtcat
     3961 tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag
     4021 ttatgtaacg cggaacgcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc
     4081 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     4141 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt
     4201 attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta
     4261 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4321 tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4381 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     4441 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
     4501 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4561 actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata
     4621 agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4681 cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg
     4741 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     4801 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     4861 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     4921 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     4981 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     5041 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     5101 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     5161 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     5221 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     5281 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     5341 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     5401 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     5461 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     5521 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     5581 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     5641 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     5701 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     5761 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtggaag
     5821 ccgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     5881 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     5941 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     6001 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     6061 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     6121 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     6181 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     6241 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     6301 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     6361 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     6421 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     6481 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     6541 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     6601 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     6661 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     6721 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     6781 gtgagggcct atttcccatg attccttcat atttgcatat acgatacaag gctgttagag
     6841 agataattgg aattaatttg actgtaaaca caaagatatt agtacaaaat acgtgacgta
     6901 gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa atggactatc
     6961 atatgcttac cgtaacttga aagtatttcg atttcttggc tttatatatc ttgtggaaag
     7021 gacgaaacac cgggtcttcg agaagacctg ttttagagct agaaatagca agttaaaata
     7081 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt tgttttagag
     7141 ctagaaatag caagttaaaa taaggctagt ccgtttttag cgcgtgcgcc aattctgcag
     7201 acaaatggct ctagaggtac ccgttacata acttacggta aatggcccgc ctggctgacc
     7261 gcccaacgac ccccgcccat tgacgtcaat agtaacgcca atagggactt tccattgacg
     7321 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
     7381 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attgtgccca
     7441 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
     7501 taccatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc ccccctcccc
     7561 acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg gggcgggggg
     7621 ggggggggsg cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga
     7681 gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc
     7741 ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgcgct
     7801 gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg ctctgactga
     7861 ccgcgttact cccacaggtg agcgggcggg acggcccttc tcctccgggc tgtaattagc
     7921 tgagcaagag gtaagggttt aagggatggt tggttggtgg ggtattaatg tttaattacc
     7981 tggagcacct gcctgaaatc actttttttc aggttggacc ggtgccacca tggactataa
     8041 ggaccacgac ggagactaca aggatcatga tattgattac aaagacgatg acgataagat
     8101 ggccccaaag aagaagcgga aggtcggtat ccacggagtc ccagcagccg acaagaagta
     8161 cagcatcggc ctggccatcg gcaccaactc tgtgggctgg gccgtgatca ccgacgagta
     8221 caaggtgccc agcaagaaat tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa
     8281 gaacctgatc ggagccctgc tgttcgacag cggcgaaaca gccgaggcca cccggctgaa
     8341 gagaaccgcc agaagaagat acaccagacg gaagaaccgg atctgctatc tgcaagagat
     8401 cttcagcaac gagatggcca aggtggacga cagcttcttc cacagactgg aagagtcctt
     8461 cctggtggaa gaggataaga agcacgagcg gcaccccatc ttcggcaaca tcgtggacga
     8521 ggtggcctac cacgagaagt accccaccat ctaccacctg agaaagaaac tggtggacag
     8581 caccgacaag gccgacctgc ggctgatcta tctggccctg gcccacatga tcaagttccg
     8641 gggccacttc ctgatcgagg gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt
     8701 catccagctg gtgcagacct acaaccagct gttcgaggaa aaccccatca acgccagcgg
     8761 cgtggacgcc aaggccatcc tgtctgccag actgagcaag agcagacggc tggaaaatct
     8821 gatcgcccag ctgcccggcg agaagaagaa tggcctgttc ggcaacctga ttgccctgag
     8881 cctgggcctg acccccaact tcaagagcaa cttcgacctg gccgaggatg ccaaactgca
     8941 gctgagcaag gacacctacg acgacgacct ggacaacctg ctggcccaga tcggcgacca
     9001 gtacgccgac ctgtttctgg ccgccaagaa cctgtccgac gccatcctgc tgagcgacat
     9061 cctgagagtg aacaccgaga tcaccaaggc ccccctgagc gcctctatga tcaagagata
     9121 cgacgagcac caccaggacc tgaccctgct gaaagctctc gtgcggcagc agctgcctga
     9181 gaagtacaaa gagattttct tcgaccagag caagaacggc tacgccggct acattgacgg
     9241 cggagccagc caggaagagt tctacaagtt catcaagccc atcctggaaa agatggacgg
     9301 caccgaggaa ctgctcgtga agctgaacag agaggacctg ctgcggaagc agcggacctt
     9361 cgacaacggc agcatccccc accagatcca cctgggagag ctgcacgcca ttctgcggcg
     9421 gcaggaagat ttttacccat tcctgaagga caaccgggaa aagatcgaga agatcctgac
     9481 cttccgcatc ccctactacg tgggccctct ggccagggga aacagcagat tcgcctggat
     9541 gaccagaaag agcgaggaaa ccatcacccc ctggaacttc gaggaagtgg tggacaaggg
     9601 cgcttccgcc cagagcttca tcgagcggat gaccaacttc gataagaacc tgcccaacga
     9661 gaaggtgctg cccaagcaca gcctgctgta cgagtacttc accgtgtata acgagctgac
     9721 caaagtgaaa tacgtgaccg agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa
     9781 aaaggccatc gtggacctgc tgttcaagac caaccggaaa gtgaccgtga agcagctgaa
     9841 agaggactac ttcaagaaaa tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga
     9901 tcggttcaac gcctccctgg gcacatacca cgatctgctg aaaattatca aggacaagga
     9961 cttcctggac aatgaggaaa acgaggacat tctggaagat atcgtgctga ccctgacact
    10021 gtttgaggac agagagatga tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga
    10081 caaagtgatg aagcagctga agcggcggag atacaccggc tggggcaggc tgagccggaa
    10141 gctgatcaac ggcatccggg acaagcagtc cggcaagaca atcctggatt tcctgaagtc
    10201 cgacggcttc gccaacagaa acttcatgca gctgatccac gacgacagcc tgacctttaa
    10261 agaggacatc cagaaagccc aggtgtccgg ccagggcgat agcctgcacg agcacattgc
    10321 caatctggcc ggcagccccg ccattaagaa gggcatcctg cagacagtga aggtggtgga
    10381 cgagctcgtg aaagtgatgg gccggcacaa gcccgagaac atcgtgatcg aaatggccag
    10441 agagaaccag accacccaga agggacagaa gaacagccgc gagagaatga agcggatcga
    10501 agagggcatc aaagagctgg gcagccagat cctgaaagaa caccccgtgg aaaacaccca
    10561 gctgcagaac gagaagctgt acctgtacta cctgcagaat gggcgggata tgtacgtgga
    10621 ccaggaactg gacatcaacc ggctgtccga ctacgatgtg gacgctatcg tgcctcagag
    10681 ctttctgaag gacgactcca tcgacaacaa ggtgctgacc agaagcgaca agaaccgggg
    10741 caagagcgac aacgtgccct ccgaagaggt cgtgaagaag atgaagaact actggcggca
    10801 gctgctgaac gccaagctga ttacccagag aaagttcgac aatctgacca aggccgagag
    10861 aggcggcctg agcgaactgg ataaggccgg cttcatcaag agacagctgg tggaaacccg
    10921 gcagatcaca aagcacgtgg cacagatcct ggactcccgg atgaacacta agtacgacga
    10981 gaatgacaag ctgatccggg aagtgaaagt gatcaccctg aagtccaagc tggtgtccga
    11041 tttccggaag gatttccagt tttacaaagt gcgcgagatc aacaactacc accacgccca
    11101 cgacgcctac ctgaacgccg tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga
    11161 aagcgagttc gtgtacggcg actacaaggt gtacgacgtg cggaagatga tcgccaagag
    11221 cgagcaggaa atcggcaagg ctaccgccaa gtacttcttc tacagcaaca tcatgaactt
    11281 tttcaagacc gagattaccc tggccaacgg cgagatccgg aagcggcctc tgatcgagac
    11341 aaacggcgaa accggggaga tcgtgtggga taagggccgg gattttgcca ccgtgcggaa
    11401 agtgctgagc atgccccaag tgaatatcgt gaaaaagacc gaggtgcaga caggcggctt
    11461 cagcaaagag tctatcctgc ccaagaggaa cagcgataag ctgatcgcca gaaagaagga
    11521 ctgggaccct aagaagtacg gcggcttcga cagccccacc gtggcctatt ctgtgctggt
    11581 ggtggccaaa gtggaaaagg gcaagtccaa gaaactgaag agtgtgaaag agctgctggg
    11641 gatcaccatc atggaaagaa gcagcttcga gaagaatccc atcgactttc tggaagccaa
    11701 gggctacaaa gaagtgaaaa aggacctgat catcaagctg c
//
