LOCUS       sequence-259366-        9990 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dCas9-m6A Writer. Methylates adenines at local GATC sites. 
            pCAG-dCas9-NLS-3xFLAG-3AC3L linker-EcoDam(N132A).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9990)
  AUTHORS   Park M, Patel N, Keung AJ, Khalil AS
  TITLE     Engineering Epigenetic Regulation Using Synthetic Read-Write 
            Modules.
  JOURNAL   Cell. 2019 Jan 10;176(1-2):227-238.e20. doi: 
            10.1016/j.cell.2018.11.002. Epub 2018 Dec 6.
  PUBMED    30528434
REFERENCE   2  (bases 1 to 9990)
  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: "Cell."; 
            date: "2019-01-10"; volume: "176(1-2):227-238.e20. doi"; pages: " 
            10.1016/j.cell.2018.11.002. Epub 2018 Dec 6"
COMMENT     Sequence Label: pMP416
FEATURES             Location/Qualifiers
     source          1..9990
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1670
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1594..1616
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1678..1697
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1763..1772
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1769..5170,5171..5173,5174..5770,5771,5772..5774,
                     5775..5776,5777..5779,5780..5872)
                     /codon_start=1
                     /product="Streptococcus pyogenes Cas9 endonuclease with the
                     D1135V/R1335Q/T1337R mutations"
                     /label=Cas9 VQR
                     /note="has an altered recognition specificity for the 
                     protospacer adjacent motif (PAM) (Kleinstiver et al., 
                     2015)"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             join(1769..1795,1796..1798,1799..4285,4286..4288,
                     4289..5872)
                     /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="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             5879..5899
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5906..5971
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(7014..7033)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    7079..7134
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(7133..7152)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(7493..7509)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7493..7509)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7506..7528)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7517..7533
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(7541..7547,7548..7565,7566..7571))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7586..7607
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        7666..7861
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7712..7847
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    7867..8001
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7904..7923)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7958..7977
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8069..8086)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8240..8828)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8320..8339)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8999..9790,9791..9859))
                     /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     9622..9641
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9860..9964)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
     1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
     1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
     1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
     1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     1681 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attctgcagt cgacggtacc
     1741 gcgggcccgg gatccaccgg tcgccaccat ggataaaaag tattctattg gtttagccat
     1801 cggcactaat tccgttggat gggctgtcat aaccgatgaa tacaaagtac cttcaaagaa
     1861 atttaaggtg ttggggaaca cagaccgtca ttcgattaaa aagaatctta tcggtgccct
     1921 cctattcgat agtggcgaaa cggcagaggc gactcgcctg aaacgaaccg ctcggagaag
     1981 gtatacacgt cgcaagaacc gaatatgtta cttacaagaa atttttagca atgagatggc
     2041 caaagttgac gattctttct ttcaccgttt ggaagagtcc ttccttgtcg aagaggacaa
     2101 gaaacatgaa cggcacccca tctttggaaa catagtagat gaggtggcat atcatgaaaa
     2161 gtacccaacg atttatcacc tcagaaaaaa gctagttgac tcaactgata aagcggacct
     2221 gaggttaatc tacttggctc ttgcccatat gataaagttc cgtgggcact ttctcattga
     2281 gggtgatcta aatccggaca actcggatgt cgacaaactg ttcatccagt tagtacaaac
     2341 ctataatcag ttgtttgaag agaaccctat aaatgcaagt ggcgtggatg cgaaggctat
     2401 tcttagcgcc cgcctctcta aatcccgacg gctagaaaac ctgatcgcac aattacccgg
     2461 agagaagaaa aatgggttgt tcggtaacct tatagcgctc tcactaggcc tgacaccaaa
     2521 ttttaagtcg aacttcgact tagctgaaga tgccaaattg cagcttagta aggacacgta
     2581 cgatgacgat ctcgacaatc tactggcaca aattggagat cagtatgcgg acttattttt
     2641 ggctgccaaa aaccttagcg atgcaatcct cctatctgac atactgagag ttaatactga
     2701 gattaccaag gcgccgttat ccgcttcaat gatcaaaagg tacgatgaac atcaccaaga
     2761 cttgacactt ctcaaggccc tagtccgtca gcaactgcct gagaaatata aggaaatatt
     2821 ctttgatcag tcgaaaaacg ggtacgcagg ttatattgac ggcggagcga gtcaagagga
     2881 attctacaag tttatcaaac ccatattaga gaagatggat gggacggaag agttgcttgt
     2941 aaaactcaat cgcgaagatc tactgcgaaa gcagcggact ttcgacaacg gtagcattcc
     3001 acatcaaatc cacttaggcg aattgcatgc tatacttaga aggcaggagg atttttatcc
     3061 gttcctcaaa gacaatcgtg aaaagattga gaaaatccta acctttcgca taccttacta
     3121 tgtgggaccc ctggcccgag ggaactctcg gttcgcatgg atgacaagaa agtccgaaga
     3181 aacgattact ccatggaatt ttgaggaagt tgtcgataaa ggtgcgtcag ctcaatcgtt
     3241 catcgagagg atgaccaact ttgacaagaa tttaccgaac gaaaaagtat tgcctaagca
     3301 cagtttactt tacgagtatt tcacagtgta caatgaactc acgaaagtta agtatgtcac
     3361 tgagggcatg cgtaaacccg cctttctaag cggagaacag aagaaagcaa tagtagatct
     3421 gttattcaag accaaccgca aagtgacagt taagcaattg aaagaggact actttaagaa
     3481 aattgaatgc ttcgattctg tcgagatctc cggggtagaa gatcgattta atgcgtcact
     3541 tggtacgtat catgacctcc taaagataat taaagataag gacttcctgg ataacgaaga
     3601 gaatgaagat atcttagaag atatagtgtt gactcttacc ctctttgaag atcgggaaat
     3661 gattgaggaa agactaaaaa catacgctca cctgttcgac gataaggtta tgaaacagtt
     3721 aaagaggcgt cgctatacgg gctggggacg attgtcgcgg aaacttatca acgggataag
     3781 agacaagcaa agtggtaaaa ctattctcga ttttctaaag agcgacggct tcgccaatag
     3841 gaactttatg cagctgatcc atgatgactc tttaaccttc aaagaggata tacaaaaggc
     3901 acaggtttcc ggacaagggg actcattgca cgaacatatt gcgaatcttg ctggttcgcc
     3961 agccatcaaa aagggcatac tccagacagt caaagtagtg gatgagctag ttaaggtcat
     4021 gggacgtcac aaaccggaaa acattgtaat cgagatggca cgcgaaaatc aaacgactca
     4081 gaaggggcaa aaaaacagtc gagagcggat gaagagaata gaagagggta ttaaagaact
     4141 gggcagccag atcttaaagg agcatcctgt ggaaaatacc caattgcaga acgagaaact
     4201 ttacctctat tacctacaaa atggaaggga catgtatgtt gatcaggaac tggacataaa
     4261 ccgtttatct gattacgacg tcgatgccat tgtaccccaa tcctttttga aggacgattc
     4321 aatcgacaat aaagtgctta cacgctcgga taagaaccga gggaaaagtg acaatgttcc
     4381 aagcgaggaa gtcgtaaaga aaatgaagaa ctattggcgg cagctcctaa atgcgaaact
     4441 gataacgcaa agaaagttcg ataacttaac taaagctgag aggggtggct tgtctgaact
     4501 tgacaaggcc ggatttatta aacgtcagct cgtggaaacc cgccaaatca caaagcatgt
     4561 tgcacagata ctagattccc gaatgaatac gaaatacgac gagaacgata agctgattcg
     4621 ggaagtcaaa gtaatcactt taaagtcaaa attggtgtcg gacttcagaa aggattttca
     4681 attctataaa gttagggaga taaataacta ccaccatgcg cacgacgctt atcttaatgc
     4741 cgtcgtaggg accgcactca ttaagaaata cccgaagcta gaaagtgagt ttgtgtatgg
     4801 tgattacaaa gtttatgacg tccgtaagat gatcgcgaaa agcgaacagg agataggcaa
     4861 ggctacagcc aaatacttct tttattctaa cattatgaat ttctttaaga cggaaatcac
     4921 tctggcaaac ggagagatac gcaaacgacc tttaattgaa accaatgggg agacaggtga
     4981 aatcgtatgg gataagggcc gggacttcgc gacggtgaga aaagttttgt ccatgcccca
     5041 agtcaacata gtaaagaaaa ctgaggtgca gaccggaggg ttttcaaagg aatcgattct
     5101 tccaaaaagg aatagtgata agctcatcgc tcgtaaaaag gactgggacc cgaaaaagta
     5161 cggtggcttc gatagcccta cagttgccta ttctgtccta gtagtggcaa aagttgagaa
     5221 gggaaaatcc aagaaactga agtcagtcaa agaattattg gggataacga ttatggagcg
     5281 ctcgtctttt gaaaagaacc ccatcgactt ccttgaggcg aaaggttaca aggaagtaaa
     5341 aaaggatctc ataattaaac taccaaagta tagtctgttt gagttagaaa atggccgaaa
     5401 acggatgttg gctagcgccg gagagcttca aaaggggaac gaactcgcac taccgtctaa
     5461 atacgtgaat ttcctgtatt tagcgtccca ttacgagaag ttgaaaggtt cacctgaaga
     5521 taacgaacag aagcaacttt ttgttgagca gcacaaacat tatctcgacg aaatcataga
     5581 gcaaatttcg gaattcagta agagagtcat cctagctgat gccaatctgg acaaagtatt
     5641 aagcgcatac aacaagcaca gggataaacc catacgtgag caggcggaaa atattatcca
     5701 tttgtttact cttaccaacc tcggcgctcc agccgcattc aagtattttg acacaacgat
     5761 agatcgcaaa cgatacactt ctaccaagga ggtgctagac gcgacactga ttcaccaatc
     5821 catcacggga ttatatgaaa ctcggataga tttgtcacag cttgggggtg acggatcccc
     5881 caagaagaag aggaaagtct cgagcgacta caaagaccat gacggtgatt ataaagatca
     5941 tgacatcgat tacaaggatg acgatgacaa ggctgcagga ggcggtggaa gcgggggaga
     6001 gctctctagt ggcaatagca acgccaactc ccggggcccc agtttttcct ccggactggt
     6061 gcccctgtcc ctgagagggt cacatgcctg caggaagaaa aatcgcgctt ttttgaagtg
     6121 ggcagggggc aagtatcccc tgcttgatga tattaaacgg catttgccca agggcgaatg
     6181 tctggttgag ccttttgtag gtgccgggtc ggtgtttctc aacaccgact tttctcgtta
     6241 tatccttgcc gatatcaata gcgacctgat cagtctctat aacattgtga agatgcgtac
     6301 tgatgagtac gtacaggccg cacgcgagct gtttgttccc gaaacaaatt gcgccgaggt
     6361 ttactatcag ttccgcgaag agttcaacaa aagccaggat ccgttccgtc gggcggtact
     6421 gtttttatat ttgaaccgct acggttacaa cggcctgtgt cgttacaatc tgcgcggtga
     6481 gtttgccgtg ccgttcggcc gctacaaaaa accctatttc ccggaagcag agttgtatca
     6541 cttcgctgaa aaagcgcaga atgccttttt ctattgtgag tcttacgccg atagcatggc
     6601 gcgcgcagat gatgcatccg tcgtctattg cgatccgcct tatgcaccgc tgtctgcgac
     6661 cgccaacttt acggcgtatc acacaaacag ttttacgctt gaacaacaag cgcatctggc
     6721 ggagatcgcc gaaggtctgg ttgagcgcca tattccagtg ctgatctcca atcacgatac
     6781 gatgttaacg cgtgagtggt atcagcgcgc aaaattgcat gtcgtcaaag ttcgacgcag
     6841 tataagcagc aacggcggca cacgtaaaaa ggtggacgaa ctgctggctt tgtacaaacc
     6901 aggagtcgtt tcacccgcga aaaaataagc ggccgcactc ctcaggtgca ggctgcctat
     6961 cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca aataccactg agatcttttt
     7021 ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac ttctggctaa
     7081 taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga
     7141 aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt ttagagtttg
     7201 gcaacatatg ccatatgctg gctgccatga acaaaggtgg ctataaagag gtcatcagta
     7261 tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga cttgaggtta
     7321 gatttttttt atattttgtt ttgtgttatt tttttcttta acatccctaa aattttcctt
     7381 acatgtttta ctagccagat ttttcctcct ctcctgacta ctcccagtca tagctgtccc
     7441 tcttctctta tgaagatccc tcgacctgca gcccaagctt ggcgtaatca tggtcatagc
     7501 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     7561 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     7621 cactgcccgc tttccagtcg ggaaacctgt cgtgccagcg gatccgcatc tcaattagtc
     7681 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     7741 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     7801 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     7861 aaagctaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     7921 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     7981 gtatcttatc atgtctggat ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     8041 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     8101 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     8161 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     8221 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     8281 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     8341 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     8401 cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt
     8461 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     8521 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     8581 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     8641 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     8701 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     8761 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     8821 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     8881 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     8941 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     9001 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     9061 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     9121 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     9181 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     9241 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     9301 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     9361 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     9421 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     9481 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     9541 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     9601 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     9661 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     9721 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     9781 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     9841 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     9901 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     9961 cgcgcacatt tccccgaaaa gtgccacctg
//
