LOCUS       sequence-267083-        9420 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of fusion protein M3D395A-M14-dCas9 in Mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9420)
  AUTHORS   Liu XM, Zhou J, Mao Y, Ji Q, Qian SB
  TITLE     Programmable RNA N(6)-methyladenosine editing by CRISPR-Cas9 
            conjugates.
  JOURNAL   Nat Chem Biol. 2019 Sep;15(9):865-871. doi: 
            10.1038/s41589-019-0327-1. Epub 2019 Aug 5.
  PUBMED    31383972
REFERENCE   2  (bases 1 to 9420)
  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"; doi: 
            "10.1038/s41589-019-0327-1"; journalName: "Nat Chem Biol"; date: 
            "2019-09"; volume: "15"; issue: "9"; pages: "865-871"
COMMENT     Sequence Label: M3D395A-M14-dCas9
FEATURES             Location/Qualifiers
     source          1..9420
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(2281..5679,5680..5682,5683..6279,6280,6281..6283,
                     6284..6285,6286..6288,6289..6381)
                     /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="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(2281..2304,2305..2307,2308..4794,4795..4797,
                     4798..6381)
                     /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             6394..6642
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6655..6675
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6684..6701
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6724..6741)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6730..6954
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7025..7041)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7025..7041)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7038..7060)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7049..7065
                     /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(7073..7079,7080..7097,7098..7103))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7118..7139
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7256..7273)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7427..8015)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7507..7526)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8186..8977,8978..9046))
                     /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     8809..8828
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9047..9151)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9230..9249)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
      601 agatccgcta gagatccgcg gccgctaata cgactcacta tagggagagc cgccaccatg
      661 ttcccacctc agtggatctg ttgtgatatc cgctacctgg acgtcagtat cttgggcaag
      721 tttgcagttg tgatggctgc cccaccctgg gatattcaca tggaactgcc ctatgggacc
      781 ctgacagatg atgagatgcg caggctcaac atacccgtac tacaggatga tggctttctc
      841 ttcctctggg tcacaggcag ggccatggag ttggggagag aatgtctaaa cctctggggg
      901 tatgaacggg tagatgaaat tatttgggtg aagacaaatc aactgcaacg catcattcgg
      961 acaggccgta caggtcactg gttgaaccat gggaaggaac actgcttggt tggtgtcaaa
     1021 ggaaatcccc aaggcttcaa ccagggtctg gattgtgatg tgatcgtagc tgaggttcgt
     1081 tccaccagtc ataaaccaga tgaaatctat ggcatgattg aaagactatc tcctggcact
     1141 cgcaagattg agttatttgg acgaccacac aatgtgcaac ccaactggat cacccttgga
     1201 aaccaactgg atgggatcca cctactagac ccagatgtgg ttgcacggtt caagcaaagg
     1261 tacccagatg gtatcatctc taaacctaag aatttaggcg gatcaggtgg ctccggaggg
     1321 tctggtggat cgggaacaca gagcttaaat ccccataatg attactgcca acattttgta
     1381 gacactggac atagacctca gaatttcatc agggatgtag gtttagctga cagatttgaa
     1441 gaatatccta aactgaggga gctcatcagg ctaaaggatg agttaatagc taaatctaac
     1501 actcctccca tgtacttaca agccgatata gaagcctttg acatcagaga actaacaccc
     1561 aaatttgatg tgattcttct ggaaccccct ttagaagaat attacagaga aactggcatc
     1621 actgctaatg aaaaatgctg gacttgggat gatattatga agttagaaat tgatgagatt
     1681 gcagcacctc gatcatttat ttttctctgg tgtggttctg gggaggggtt ggaccttgga
     1741 agagtgtgtt tacgaaaatg gggttacaga agatgtgaag atatttgttg gattaaaacc
     1801 aataaaaaca atcctgggaa gactaagact ttagatccaa aggctgtctt tcagagaaca
     1861 aaggaacact gcctcatggg gatcaaagga actgtgaagc gtagcacaga cggggacttc
     1921 attcatgcta atgttgacat tgacttaatt atcacagaag aacctgaaat tggcaatata
     1981 gaaaaacctg tagaaatttt tcatataatt gagcattttt gtcttggtag aagacgcctt
     2041 catctatttg gaagagatag tacaattcga ccaggctggc tcacagttgg accaacgctt
     2101 acaaatagca actacaatgc agaaacatat gcatcctatt tcagtgctcc taattcctac
     2161 ttgactggtt gtacagaaga aattgagaga cttcgaccaa aatcgcctcc tcccaaatct
     2221 aaatctgacc gaagcggcag cgagactccc gggacctcag agtccgccac acccgaaagt
     2281 gataaaaagt attctattgg tttagccatc ggcactaatt ccgttggatg ggctgtcata
     2341 accgatgaat acaaagtacc ttcaaagaaa tttaaggtgt tggggaacac agaccgtcat
     2401 tcgattaaaa agaatcttat cggtgccctc ctattcgata gtggcgaaac ggcagaggcg
     2461 actcgcctga aacgaaccgc tcggagaagg tatacacgtc gcaagaaccg aatatgttac
     2521 ttacaagaaa tttttagcaa tgagatggcc aaagttgacg attctttctt tcaccgtttg
     2581 gaagagtcct tccttgtcga agaggacaag aaacatgaac ggcaccccat ctttggaaac
     2641 atagtagatg aggtggcata tcatgaaaag tacccaacga tttatcacct cagaaaaaag
     2701 ctagttgact caactgataa agcggacctg aggttaatct acttggctct tgcccatatg
     2761 ataaagttcc gtgggcactt tctcattgag ggtgatctaa atccggacaa ctcggatgtc
     2821 gacaaactgt tcatccagtt agtacaaacc tataatcagt tgtttgaaga gaaccctata
     2881 aatgcaagtg gcgtggatgc gaaggctatt cttagcgccc gcctctctaa atcccgacgg
     2941 ctagaaaacc tgatcgcaca attacccgga gagaagaaaa atgggttgtt cggtaacctt
     3001 atagcgctct cactaggcct gacaccaaat tttaagtcga acttcgactt agctgaagat
     3061 gccaaattgc agcttagtaa ggacacgtac gatgacgatc tcgacaatct actggcacaa
     3121 attggagatc agtatgcgga cttatttttg gctgccaaaa accttagcga tgcaatcctc
     3181 ctatctgaca tactgagagt taatactgag attaccaagg cgccgttatc cgcttcaatg
     3241 atcaaaaggt acgatgaaca tcaccaagac ttgacacttc tcaaggccct agtccgtcag
     3301 caactgcctg agaaatataa ggaaatattc tttgatcagt cgaaaaacgg gtacgcaggt
     3361 tatattgacg gcggagcgag tcaagaggaa ttctacaagt ttatcaaacc catattagag
     3421 aagatggatg ggacggaaga gttgcttgta aaactcaatc gcgaagatct actgcgaaag
     3481 cagcggactt tcgacaacgg tagcattcca catcaaatcc acttaggcga attgcatgct
     3541 atacttagaa ggcaggagga tttttatccg ttcctcaaag acaatcgtga aaagattgag
     3601 aaaatcctaa cctttcgcat accttactat gtgggacccc tggcccgagg gaactctcgg
     3661 ttcgcatgga tgacaagaaa gtccgaagaa acgattactc catggaattt tgaggaagtt
     3721 gtcgataaag gtgcgtcagc tcaatcgttc atcgagagga tgaccaactt tgacaagaat
     3781 ttaccgaacg aaaaagtatt gcctaagcac agtttacttt acgagtattt cacagtgtac
     3841 aatgaactca cgaaagttaa gtatgtcact gagggcatgc gtaaacccgc ctttctaagc
     3901 ggagaacaga agaaagcaat agtagatctg ttattcaaga ccaaccgcaa agtgacagtt
     3961 aagcaattga aagaggacta ctttaagaaa attgaatgct tcgattctgt cgagatctcc
     4021 ggggtagaag atcgatttaa tgcgtcactt ggtacgtatc atgacctcct aaagataatt
     4081 aaagataagg acttcctgga taacgaagag aatgaagata tcttagaaga tatagtgttg
     4141 actcttaccc tctttgaaga tcgggaaatg attgaggaaa gactaaaaac atacgctcac
     4201 ctgttcgacg ataaggttat gaaacagtta aagaggcgtc gctatacggg ctggggacga
     4261 ttgtcgcgga aacttatcaa cgggataaga gacaagcaaa gtggtaaaac tattctcgat
     4321 tttctaaaga gcgacggctt cgccaatagg aactttatgc agctgatcca tgatgactct
     4381 ttaaccttca aagaggatat acaaaaggca caggtttccg gacaagggga ctcattgcac
     4441 gaacatattg cgaatcttgc tggttcgcca gccatcaaaa agggcatact ccagacagtc
     4501 aaagtagtgg atgagctagt taaggtcatg ggacgtcaca aaccggaaaa cattgtaatc
     4561 gagatggcac gcgaaaatca aacgactcag aaggggcaaa aaaacagtcg agagcggatg
     4621 aagagaatag aagagggtat taaagaactg ggcagccaga tcttaaagga gcatcctgtg
     4681 gaaaataccc aattgcagaa cgagaaactt tacctctatt acctacaaaa tggaagggac
     4741 atgtatgttg atcaggaact ggacataaac cgtttatctg attacgacgt cgatgccatt
     4801 gtaccccaat cctttttgaa ggacgattca atcgacaata aagtgcttac acgctcggat
     4861 aagaaccgag ggaaaagtga caatgttcca agcgaggaag tcgtaaagaa aatgaagaac
     4921 tattggcggc agctcctaaa tgcgaaactg ataacgcaaa gaaagttcga taacttaact
     4981 aaagctgaga ggggtggctt gtctgaactt gacaaggccg gatttattaa acgtcagctc
     5041 gtggaaaccc gccaaatcac aaagcatgtt gcacagatac tagattcccg aatgaatacg
     5101 aaatacgacg agaacgataa gctgattcgg gaagtcaaag taatcacttt aaagtcaaaa
     5161 ttggtgtcgg acttcagaaa ggattttcaa ttctataaag ttagggagat aaataactac
     5221 caccatgcgc acgacgctta tcttaatgcc gtcgtaggga ccgcactcat taagaaatac
     5281 ccgaagctag aaagtgagtt tgtgtatggt gattacaaag tttatgacgt ccgtaagatg
     5341 atcgcgaaaa gcgaacagga gataggcaag gctacagcca aatacttctt ttattctaac
     5401 attatgaatt tctttaagac ggaaatcact ctggcaaacg gagagatacg caaacgacct
     5461 ttaattgaaa ccaatgggga gacaggtgaa atcgtatggg ataagggccg ggacttcgcg
     5521 acggtgagaa aagttttgtc catgccccaa gtcaacatag taaagaaaac tgaggtgcag
     5581 accggagggt tttcaaagga atcgattctt ccaaaaagga atagtgataa gctcatcgct
     5641 cgtaaaaagg actgggaccc gaaaaagtac ggtggcttcg atagccctac agttgcctat
     5701 tctgtcctag tagtggcaaa agttgagaag ggaaaatcca agaaactgaa gtcagtcaaa
     5761 gaattattgg ggataacgat tatggagcgc tcgtcttttg aaaagaaccc catcgacttc
     5821 cttgaggcga aaggttacaa ggaagtaaaa aaggatctca taattaaact accaaagtat
     5881 agtctgtttg agttagaaaa tggccgaaaa cggatgttgg ctagcgccgg agagcttcaa
     5941 aaggggaacg aactcgcact accgtctaaa tacgtgaatt tcctgtattt agcgtcccat
     6001 tacgagaagt tgaaaggttc acctgaagat aacgaacaga agcaactttt tgttgagcag
     6061 cacaaacatt atctcgacga aatcatagag caaatttcgg aattcagtaa gagagtcatc
     6121 ctagctgatg ccaatctgga caaagtatta agcgcataca acaagcacag ggataaaccc
     6181 atacgtgagc aggcggaaaa tattatccat ttgtttactc ttaccaacct cggcgctcca
     6241 gccgcattca agtattttga cacaacgata gatcgcaaac gatacacttc taccaaggag
     6301 gtgctagacg cgacactgat tcaccaatcc atcacgggat tatatgaaac tcggatagat
     6361 ttgtcacagc ttgggggtga ctctggtggt tctactaatc tgtcagatat tattgaaaag
     6421 gagaccggta agcaactggt tatccaggaa tccatcctca tgctcccaga ggaggtggaa
     6481 gaagtcattg ggaacaagcc ggaaagcgat atactcgtgc acaccgccta cgacgagagc
     6541 accgacgaga atgtcatgct tctgactagc gacgcccctg aatacaagcc ttgggctctg
     6601 gtcatacagg atagcaacgg tgagaacaag attaagatgc tctctggtgg ttctcccaag
     6661 aagaagagga aagtctaacc ggtcatcatc accatcacca ttgagtttaa acccgctgat
     6721 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     6781 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     6841 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     6901 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     6961 aggcggaaag aaccagctgg ggctcgatac cgtcgacctc tagctagagc ttggcgtaat
     7021 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     7081 gagccggaag cataaagtgt aaagcctagg gtgcctaatg agtgagctaa ctcacattaa
     7141 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     7201 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     7261 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     7321 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     7381 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     7441 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     7501 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     7561 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     7621 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     7681 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     7741 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     7801 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     7861 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     7921 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     7981 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     8041 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     8101 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     8161 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     8221 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     8281 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     8341 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     8401 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     8461 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     8521 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     8581 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     8641 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     8701 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     8761 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     8821 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     8881 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     8941 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     9001 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     9061 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     9121 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     9181 tcgacggatc gggagatcga tctcccgatc ccctagggtc gactctcagt acaatctgct
     9241 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
     9301 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
     9361 atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt
//
