LOCUS       sequence-438193-       11743 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses dCAS9- R882H Mutated DNMT3A fusion protein and double 
            marker GFP-T2A-Puro under independent promoters.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11743)
  TITLE     Systemic comparison of dCas9-based DNA methylation editing systems 
            for specificity and stability
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 11743)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: M3-dCAS9-mutDNMT3A(R882H)(dCas9-mut3A)
FEATURES             Location/Qualifiers
     source          1..11743
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    610..731
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(647..666)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     701..720
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   745..885
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   745..874
                     /label=AAV2 ITR
     rep_origin      960..1415
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1047..1066)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1257..1278
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1432..1451)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1551..1573
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1611..1629)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1697..1801
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1802..1870,1871..2662)
                     /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(2020..2039)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2833..3421
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3322..3341
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3483..3731
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     3483..3503
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     3654..3673
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        3750..3825
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        3922..4207
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     intron          4487..4714
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      4726..4735
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4735..4800
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4807..4827
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(4852..4875,4876..4878,4879..7365,7366..7368,
                     7369..8952)
                     /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             4852..8952
                     /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             8953..9000
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             9028..9960
                     /codon_start=1
                     /product="catalytic domain of DNA methyltransferase 3A"
                     /label=DNMT3A catalytic domain
                     /translation="PSRLQMFFANNHDQEFDPPKVYPPVPAEKRKPIRVLSLFDGIATG
                     LLVLKDLGIQVGRYIASEVCEDSITVGMVRHQGKIMYVGDVRSVTQKHIQEWGPFDLVI
                     GGSPCNDLSIVNPARKGLYEGTGRLFFEFYRLLHDARPKEGDDRPFFWLFENVVAMGVS
                     DKRDISRFLESNPVMIDAKEVSAAHRARYFWGNLPGMNRPLASTVNDKLELQECLEHGR
                     IAKFSKVRTITTRSNSIKQGKDQHFPVFMNEKEDILWCTEMERVFGFPVHYTDVSNMSH
                     LARQRLLGRSWSVPVIRHLFAPLKEYFACV"
     primer_bind     complement(10005..10022)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10011..10218
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        10233..10536
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        10537..10740
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     10691..10711
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      10779..10788
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(10785..10787,10788..10790,10791..11501)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(10785..10787,10788..10790,10791..11501)
                     /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(11091..11110)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     11438..11459
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             11532..11585
                     /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             join(11586..11743,1..442)
                     /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     complement(11586..11605)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
ORIGIN
        1 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
       61 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
      121 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
      181 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga
      241 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
      301 cgccggggtg cccgccttcc tggagacctc cgcgccccac aacctcccct tctacgagcg
      361 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
      421 gacccgcaag cccggtgcct gaaagcttcg aattctgcag tcgacggtac cgcgggcccg
      481 ggatccaccg gatctagata actgatcata atcagccata ccacatttgt agaggtttta
      541 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
      601 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
      661 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
      721 aatgtatctt aacgcggcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc
      781 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
      841 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt
      901 attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta
      961 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     1021 tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     1081 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     1141 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
     1201 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     1261 actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata
     1321 agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     1381 cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg
     1441 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     1501 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     1561 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     1621 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     1681 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     1741 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     1801 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     1861 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     1921 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     1981 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     2041 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     2101 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     2161 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     2221 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     2281 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     2341 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     2401 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     2461 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtggaag
     2521 ccgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     2581 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     2641 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     2701 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     2761 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     2821 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     2881 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     2941 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     3001 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3061 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3121 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3181 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     3241 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     3301 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     3361 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     3421 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     3481 gtgagggcct atttcccatg attccttcat atttgcatat acgatacaag gctgttagag
     3541 agataattgg aattaatttg actgtaaaca caaagatatt agtacaaaat acgtgacgta
     3601 gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa atggactatc
     3661 atatgcttac cgtaacttga aagtatttcg atttcttggc tttatatatc ttgtggaaag
     3721 gacgaaacac cgggtcttcg agaagacctg ttttagagct agaaatagca agttaaaata
     3781 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt tgttttagag
     3841 ctagaaatag caagttaaaa taaggctagt ccgtttttag cgcgtgcgcc aattctgcag
     3901 acaaatggct ctagaggtac ccgttacata acttacggta aatggcccgc ctggctgacc
     3961 gcccaacgac ccccgcccat tgacgtcaat agtaacgcca atagggactt tccattgacg
     4021 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
     4081 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attgtgccca
     4141 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
     4201 taccatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc ccccctcccc
     4261 acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg gggcgggggg
     4321 gggggggggg ggcgccgggc gggggggggg gggggggggg gggggggggg gggggggggg
     4381 ggggggggcg gcggcagcca acaggagcgg cgcgctccga aagtttcctt ttatggcgag
     4441 gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg
     4501 ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact
     4561 gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta
     4621 gctgagcaag aggtaagggt ttaagggatg gttggttggt ggggtattaa tgtttaatta
     4681 cctggagcac ctgcctgaaa tcactttttt tcaggttgga ccggtgccac catggactat
     4741 aaggaccacg acggagacta caaggatcat gatattgatt acaaagacga tgacgataag
     4801 atggccccaa agaagaagcg gaaggtcggt atccacggag tcccagcagc cgacaagaag
     4861 tacagcatcg gcctggccat cggcaccaac tctgtgggct gggccgtgat caccgacgag
     4921 tacaaggtgc ccagcaagaa attcaaggtg ctgggcaaca ccgaccggca cagcatcaag
     4981 aagaacctga tcggagccct gctgttcgac agcggcgaaa cagccgaggc cacccggctg
     5041 aagagaaccg ccagaagaag atacaccaga cggaagaacc ggatctgcta tctgcaagag
     5101 atcttcagca acgagatggc caaggtggac gacagcttct tccacagact ggaagagtcc
     5161 ttcctggtgg aagaggataa gaagcacgag cggcacccca tcttcggcaa catcgtggac
     5221 gaggtggcct accacgagaa gtaccccacc atctaccacc tgagaaagaa actggtggac
     5281 agcaccgaca aggccgacct gcggctgatc tatctggccc tggcccacat gatcaagttc
     5341 cggggccact tcctgatcga gggcgacctg aaccccgaca acagcgacgt ggacaagctg
     5401 ttcatccagc tggtgcagac ctacaaccag ctgttcgagg aaaaccccat caacgccagc
     5461 ggcgtggacg ccaaggccat cctgtctgcc agactgagca agagcagacg gctggaaaat
     5521 ctgatcgccc agctgcccgg cgagaagaag aatggcctgt tcggcaacct gattgccctg
     5581 agcctgggcc tgacccccaa cttcaagagc aacttcgacc tggccgagga tgccaaactg
     5641 cagctgagca aggacaccta cgacgacgac ctggacaacc tgctggccca gatcggcgac
     5701 cagtacgccg acctgtttct ggccgccaag aacctgtccg acgccatcct gctgagcgac
     5761 atcctgagag tgaacaccga gatcaccaag gcccccctga gcgcctctat gatcaagaga
     5821 tacgacgagc accaccagga cctgaccctg ctgaaagctc tcgtgcggca gcagctgcct
     5881 gagaagtaca aagagatttt cttcgaccag agcaagaacg gctacgccgg ctacattgac
     5941 ggcggagcca gccaggaaga gttctacaag ttcatcaagc ccatcctgga aaagatggac
     6001 ggcaccgagg aactgctcgt gaagctgaac agagaggacc tgctgcggaa gcagcggacc
     6061 ttcgacaacg gcagcatccc ccaccagatc cacctgggag agctgcacgc cattctgcgg
     6121 cggcaggaag atttttaccc attcctgaag gacaaccggg aaaagatcga gaagatcctg
     6181 accttccgca tcccctacta cgtgggccct ctggccaggg gaaacagcag attcgcctgg
     6241 atgaccagaa agagcgagga aaccatcacc ccctggaact tcgaggaagt ggtggacaag
     6301 ggcgcttccg cccagagctt catcgagcgg atgaccaact tcgataagaa cctgcccaac
     6361 gagaaggtgc tgcccaagca cagcctgctg tacgagtact tcaccgtgta taacgagctg
     6421 accaaagtga aatacgtgac cgagggaatg agaaagcccg ccttcctgag cggcgagcag
     6481 aaaaaggcca tcgtggacct gctgttcaag accaaccgga aagtgaccgt gaagcagctg
     6541 aaagaggact acttcaagaa aatcgagtgc ttcgactccg tggaaatctc cggcgtggaa
     6601 gatcggttca acgcctccct gggcacatac cacgatctgc tgaaaattat caaggacaag
     6661 gacttcctgg acaatgagga aaacgaggac attctggaag atatcgtgct gaccctgaca
     6721 ctgtttgagg acagagagat gatcgaggaa cggctgaaaa cctatgccca cctgttcgac
     6781 gacaaagtga tgaagcagct gaagcggcgg agatacaccg gctggggcag gctgagccgg
     6841 aagctgatca acggcatccg ggacaagcag tccggcaaga caatcctgga tttcctgaag
     6901 tccgacggct tcgccaacag aaacttcatg cagctgatcc acgacgacag cctgaccttt
     6961 aaagaggaca tccagaaagc ccaggtgtcc ggccagggcg atagcctgca cgagcacatt
     7021 gccaatctgg ccggcagccc cgccattaag aagggcatcc tgcagacagt gaaggtggtg
     7081 gacgagctcg tgaaagtgat gggccggcac aagcccgaga acatcgtgat cgaaatggcc
     7141 agagagaacc agaccaccca gaagggacag aagaacagcc gcgagagaat gaagcggatc
     7201 gaagagggca tcaaagagct gggcagccag atcctgaaag aacaccccgt ggaaaacacc
     7261 cagctgcaga acgagaagct gtacctgtac tacctgcaga atgggcggga tatgtacgtg
     7321 gaccaggaac tggacatcaa ccggctgtcc gactacgatg tggacgctat cgtgcctcag
     7381 agctttctga aggacgactc catcgacaac aaggtgctga ccagaagcga caagaaccgg
     7441 ggcaagagcg acaacgtgcc ctccgaagag gtcgtgaaga agatgaagaa ctactggcgg
     7501 cagctgctga acgccaagct gattacccag agaaagttcg acaatctgac caaggccgag
     7561 agaggcggcc tgagcgaact ggataaggcc ggcttcatca agagacagct ggtggaaacc
     7621 cggcagatca caaagcacgt ggcacagatc ctggactccc ggatgaacac taagtacgac
     7681 gagaatgaca agctgatccg ggaagtgaaa gtgatcaccc tgaagtccaa gctggtgtcc
     7741 gatttccgga aggatttcca gttttacaaa gtgcgcgaga tcaacaacta ccaccacgcc
     7801 cacgacgcct acctgaacgc cgtcgtggga accgccctga tcaaaaagta ccctaagctg
     7861 gaaagcgagt tcgtgtacgg cgactacaag gtgtacgacg tgcggaagat gatcgccaag
     7921 agcgagcagg aaatcggcaa ggctaccgcc aagtacttct tctacagcaa catcatgaac
     7981 tttttcaaga ccgagattac cctggccaac ggcgagatcc ggaagcggcc tctgatcgag
     8041 acaaacggcg aaaccgggga gatcgtgtgg gataagggcc gggattttgc caccgtgcgg
     8101 aaagtgctga gcatgcccca agtgaatatc gtgaaaaaga ccgaggtgca gacaggcggc
     8161 ttcagcaaag agtctatcct gcccaagagg aacagcgata agctgatcgc cagaaagaag
     8221 gactgggacc ctaagaagta cggcggcttc gacagcccca ccgtggccta ttctgtgctg
     8281 gtggtggcca aagtggaaaa gggcaagtcc aagaaactga agagtgtgaa agagctgctg
     8341 gggatcacca tcatggaaag aagcagcttc gagaagaatc ccatcgactt tctggaagcc
     8401 aagggctaca aagaagtgaa aaaggacctg atcatcaagc tgcctaagta ctccctgttc
     8461 gagctggaaa acggccggaa gagaatgctg gcctctgccg gcgaactgca gaagggaaac
     8521 gaactggccc tgccctccaa atatgtgaac ttcctgtacc tggccagcca ctatgagaag
     8581 ctgaagggct cccccgagga taatgagcag aaacagctgt ttgtggaaca gcacaagcac
     8641 tacctggacg agatcatcga gcagatcagc gagttctcca agagagtgat cctggccgac
     8701 gctaatctgg acaaagtgct gtccgcctac aacaagcacc gggataagcc catcagagag
     8761 caggccgaga atatcatcca cctgtttacc ctgaccaatc tgggagcccc tgccgccttc
     8821 aagtactttg acaccaccat cgaccggaag aggtacacca gcaccaaaga ggtgctggac
     8881 gccaccctga tccaccagag catcaccggc ctgtacgaga cacggatcga cctgtctcag
     8941 ctgggaggcg acaaaaggcc ggcggccacg aaaaaggccg gacaggccaa aaagaaaaag
     9001 ctcgagggcg gaggcgggag cggatccccc tcccggctcc agatgttctt cgctaataac
     9061 cacgaccagg aatttgaccc tccaaaggtt tacccacctg tcccagctga gaagaggaag
     9121 cccatccggg tgctgtctct ctttgatgga atcgctacag ggctcctggt gctgaaggac
     9181 ttgggcattc aggtgggccg ctacattgcc tcggaggtgt gtgaggactc catcacggtg
     9241 ggcatggtgc ggcaccaggg gaagatcatg tacgtcgggg acgtccgcag cgtcacacag
     9301 aagcatatcc aggagtgggg cccattcgat ctggtgattg ggggcagtcc ctgcaatgac
     9361 ctctccatcg tcaaccctgc tcgcaagggc ctctacgagg gcactggccg gctcttcttt
     9421 gagttctacc gcctcctgca tgatgcgcgg cccaaggagg gagatgatcg ccccttcttc
     9481 tggctctttg agaatgtggt ggccatgggc gttagtgaca agagggacat ctcgcgattt
     9541 ctcgagtcca accctgtgat gattgatgcc aaagaagtgt cagctgcaca cagggcccgc
     9601 tacttctggg gtaaccttcc cggtatgaac aggccgttgg catccactgt gaatgataag
     9661 ctggagctgc aggagtgtct ggagcatggc aggatagcca agttcagcaa agtgaggacc
     9721 attactacga ggtcaaactc cataaagcag ggcaaagacc agcattttcc tgtgttcatg
     9781 aatgagaaag aggacatctt atggtgcact gaaatggaaa gggtatttgg tttcccagtc
     9841 cactatactg acgtctccaa catgagccac ttggcgaggc agagactgct gggccggtca
     9901 tggagcgtgc cagtcatccg ccacctcttc gctccgctga aggagtattt tgcgtgtgtg
     9961 tccggccggc ccgaattcaa ttctaactag agctcgctga tcagcctcga ctgtgccttc
    10021 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
    10081 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
    10141 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagagaa
    10201 tagcaggcat gctggggagc ggccgcgttc cgcgttacat aacttacggt aaatggcccg
    10261 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
    10321 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
    10381 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
    10441 ggtaaatggc ccgcctggca ttgtgcccag tacatgacct tatgggactt tcctacttgg
    10501 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
    10561 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
    10621 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
    10681 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
    10741 ggtttagtga accgtcagat ccgctagcgc taccggtcgc caccatggtg agcaagggcg
    10801 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
    10861 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
    10921 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
    10981 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
    11041 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
    11101 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
    11161 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
    11221 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
    11281 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
    11341 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
    11401 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
    11461 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtccggactc agatctcgag
    11521 ctcaagcttc tgagggcaga ggaagtctgc taacatgcgg tgacgtcgag gagaatcctg
    11581 gcccaatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg
    11641 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc
    11701 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact ctt
//
