LOCUS       sequence-262453-        9272 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HypaCas9 nuclease in mammalian cells. Used to modify 
            genome of mouse embroys.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9272)
  AUTHORS   Ikeda A, Fujii W, Sugiura K, Naito K
  TITLE     High-fidelity endonuclease variant HypaCas9 facilitates accurate 
            allele-specific gene modification in mouse zygotes.
  JOURNAL   Commun Biol. 2019 Oct 10;2:371. doi: 10.1038/s42003-019-0627-8. 
            eCollection 2019.
  PUBMED    31633062
REFERENCE   2  (bases 1 to 9272)
  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: "Commun 
            Biol."; date: "2019-10-10"; volume: "2:371. doi"; pages: " 
            10.1038/s42003-019-0627-8. eCollection 2019"
COMMENT     Sequence Label: pCAG-T3-HypaCAS9-pA
FEATURES             Location/Qualifiers
     source          1..9272
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             80..103
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             113..133
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(158..2227,2228..2230,2231..2233,2234..2239,2240..2245,
                     2246..2248,2249..4258)
                     /codon_start=1
                     /product="Cas9 endonuclease from the Streptococcus pyogenes
                     Type II CRISPR/Cas system, mutated to improve targeting 
                     specificity without compromising on-target activity (Chen 
                     et al., 2017)"
                     /label=HypaCas9
                     /note="carries the mutations N692A, M694A, Q695A, and 
                     H698A"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRAFAA
                     LIADDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             158..4258
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRAFAA
                     LIADDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4271..4291
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4579..4598)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4644..4699
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4698..4717)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5060..5076)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5060..5076)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5073..5095)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5084..5100
                     /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(5108..5114,5115..5132,5133..5138))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5153..5174
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5233..5428
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5279..5414
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5434..5568
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5471..5490)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5525..5544
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5636..5653)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5807..6395)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5887..5906)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6566..7357,7358..7426))
                     /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     7189..7208
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7427..7531)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        7562..7941
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7943..8199
                     /label=chicken beta-actin promoter
                     /note="contains a 21-bp deletion relative to the standard 
                     chicken beta-actin promoter"
     intron          8200..9216
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     9140..9162
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     9224..9243
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
ORIGIN
        1 aattaaccct cactaaaggg aacaaaagct ggagctccac cgcggtggcg gccgctctag
       61 cccgggcgga tccaccatgg attacaagga tgacgacgat aagatcatgg ccccaaagaa
      121 gaagcggaag gtcggtatcc acggagtccc agcagccgac aagaagtact ccattgggct
      181 cgatatcggc acaaacagcg tcggctgggc cgtcattacg gacgagtaca aggtgccgag
      241 caaaaaattc aaagttctgg gcaataccga tcgccacagc ataaagaaga acctcattgg
      301 cgccctcctg ttcgactccg gggagacggc cgaagccacg cggctcaaaa gaacagcacg
      361 gcgcagatat acccgcagaa agaatcggat ctgctacctg caggagatct ttagtaatga
      421 gatggctaag gtggatgact ctttcttcca taggctggag gagtcctttt tggtggagga
      481 ggataaaaag cacgagcgcc acccaatctt tggcaatatc gtggacgagg tggcgtacca
      541 tgaaaagtac ccaaccatat atcatctgag gaagaagctt gtagacagta ctgataaggc
      601 tgacttgcgg ttgatctatc tcgcgctggc gcatatgatc aaatttcggg gacacttcct
      661 catcgagggg gacctgaacc cagacaacag cgatgtcgac aaactcttta tccaactggt
      721 tcagacttac aatcagcttt tcgaagagaa cccgatcaac gcatccggag ttgacgccaa
      781 agcaatcctg agcgctaggc tgtccaaatc ccggcggctc gaaaacctca tcgcacagct
      841 ccctggggag aagaagaacg gcctgtttgg taatcttatc gccctgtcac tcgggctgac
      901 ccccaacttt aaatctaact tcgacctggc cgaagatgcc aagcttcaac tgagcaaaga
      961 cacctacgat gatgatctcg acaatctgct ggcccagatc ggcgaccagt acgcagacct
     1021 ttttttggcg gcaaagaacc tgtcagacgc cattctgctg agtgatattc tgcgagtgaa
     1081 cacggagatc accaaagctc cgctgagcgc tagtatgatc aagcgctatg atgagcacca
     1141 ccaagacttg actttgctga aggcccttgt cagacagcaa ctgcctgaga agtacaagga
     1201 aattttcttc gatcagtcta aaaatggcta cgccggatac attgacggcg gagcaagcca
     1261 ggaggaattt tacaaattta ttaagcccat cttggaaaaa atggacggca ccgaggagct
     1321 gctggtaaag cttaacagag aagatctgtt gcgcaaacag cgcactttcg acaatggaag
     1381 catcccccac cagattcacc tgggcgaact gcacgctatc ctcaggcggc aagaggattt
     1441 ctaccccttt ttgaaagata acagggaaaa gattgagaaa atcctcacat ttcggatacc
     1501 ctactatgta ggccccctcg cccggggaaa ttccagattc gcgtggatga ctcgcaaatc
     1561 agaagagacc atcactccct ggaacttcga ggaagtcgtg gataaggggg cctctgccca
     1621 gtccttcatc gaaaggatga ctaactttga taaaaatctg cctaacgaaa aggtgcttcc
     1681 taaacactct ctgctgtacg agtacttcac agtttataac gagctcacca aggtcaaata
     1741 cgtcacagaa gggatgagaa agccagcatt cctgtctgga gagcagaaga aagctatcgt
     1801 ggacctcctc ttcaagacga accggaaagt taccgtgaaa cagctcaaag aagactattt
     1861 caaaaagatt gaatgtttcg actctgttga aatcagcgga gtggaggatc gcttcaacgc
     1921 atccctggga acgtatcacg atctcctgaa aatcattaaa gacaaggact tcctggacaa
     1981 tgaggagaac gaggacattc ttgaggacat tgtcctcacc cttacgttgt ttgaagatag
     2041 ggagatgatt gaagaacgct tgaaaactta cgctcatctc ttcgacgaca aagtcatgaa
     2101 acagctcaag aggcgccgat atacaggatg ggggcggctg tcaagaaaac tgatcaatgg
     2161 gatccgagac aagcagagtg gaaagacaat cctggatttt cttaagtccg atggatttgc
     2221 caaccgggcc ttcgctgcat tgatcgctga tgactctctc acctttaagg aggacatcca
     2281 gaaagcacaa gtttctggcc agggggacag tcttcacgag cacatcgcta atcttgcagg
     2341 tagcccagct atcaaaaagg gaatactgca gaccgttaag gtcgtggatg aactcgtcaa
     2401 agtaatggga aggcataagc ccgagaatat cgttatcgag atggcccgag agaaccaaac
     2461 tacccagaag ggacagaaga acagtaggga aaggatgaag aggattgaag agggtataaa
     2521 agaactgggg tcccaaatcc ttaaggaaca cccagttgaa aacacccagc ttcagaatga
     2581 gaagctctac ctgtactacc tgcagaacgg cagggacatg tacgtggatc aggaactgga
     2641 catcaatcgg ctctccgact acgacgtgga tcatatcgtg ccccagtctt ttctcaaaga
     2701 tgattctatt gataataaag tgttgacaag atccgataaa aatagaggga agagtgataa
     2761 cgtcccctca gaagaagttg tcaagaaaat gaaaaattat tggcggcagc tgctgaacgc
     2821 caaactgatc acacaacgga agttcgataa tctgactaag gctgaacgag gtggcctgtc
     2881 tgagttggat aaagccggct tcatcaaaag gcagcttgtt gagacacgcc agatcaccaa
     2941 gcacgtggcc caaattctcg attcacgcat gaacaccaag tacgatgaaa atgacaaact
     3001 gattcgagag gtgaaagtta ttactctgaa gtctaagctg gtctcagatt tcagaaagga
     3061 ctttcagttt tataaggtga gagagatcaa caattaccac catgcgcatg atgcctacct
     3121 gaatgcagtg gtaggcactg cacttatcaa aaaatatccc aagcttgaat ctgaatttgt
     3181 ttacggagac tataaagtgt acgatgttag gaaaatgatc gcaaagtctg agcaggaaat
     3241 aggcaaggcc accgctaagt acttctttta cagcaatatt atgaattttt tcaagaccga
     3301 gattacactg gccaatggag agattcggaa gcgaccactt atcgaaacaa acggagaaac
     3361 aggagaaatc gtgtgggaca agggtaggga tttcgcgaca gtccggaagg tcctgtccat
     3421 gccgcaggtg aacatcgtta aaaagaccga agtacagacc ggaggcttct ccaaggaaag
     3481 tatcctcccg aaaaggaaca gcgacaagct gatcgcacgc aaaaaagatt gggaccccaa
     3541 gaaatacggc ggattcgatt ctcctacagt cgcttacagt gtactggttg tggccaaagt
     3601 ggagaaaggg aagtctaaaa aactcaaaag cgtcaaggaa ctgctgggca tcacaatcat
     3661 ggagcgatca agcttcgaaa aaaaccccat cgactttctc gaggcgaaag gatataaaga
     3721 ggtcaaaaaa gacctcatca ttaagcttcc caagtactct ctctttgagc ttgaaaacgg
     3781 ccggaaacga atgctcgcta gtgcgggcga gctgcagaaa ggtaacgagc tggcactgcc
     3841 ctctaaatac gttaatttct tgtatctggc cagccactat gaaaagctca aagggtctcc
     3901 cgaagataat gagcagaagc agctgttcgt ggaacaacac aaacactacc ttgatgagat
     3961 catcgagcaa ataagcgaat tctccaaaag agtgatcctc gccgacgcta acctcgataa
     4021 ggtgctttct gcttacaata agcacaggga taagcccatc agggagcagg cagaaaacat
     4081 tatccacttg tttactctga ccaacttggg cgcgcctgca gccttcaagt acttcgacac
     4141 caccatagac agaaagcggt acacctctac aaaggaggtc ctggacgcca cactgattca
     4201 tcagtcaatt acggggctct atgaaacaag aatcgacctc tctcagctcg gtggagacag
     4261 cagggctgac cccaagaaga agaggaaggt gtgaatcgat ggtacctatg ggcaccaaag
     4321 aacctgtaaa cgttatcttt ttaaattgaa tgtgcacaaa taaaagwwwr raaaaraaaa
     4381 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     4441 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa actcgaggta gcatgcaagt gtacggaatt
     4501 cactcctcag gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca atgccctggc
     4561 tcacaaatac cactgagatc tttttccctc tgccaaaaat tatggggaca tcatgaagcc
     4621 ccttgagcat ctgacttctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg
     4681 gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt taaaacatca
     4741 gaatgagtat ttggtttaga gtttggcaac atatgcccat atgctggctg ccatgaacaa
     4801 aggttggcta taaagaggtc atcagtatat gaaacagccc cctgctgtcc attccttatt
     4861 ccatagaaaa gccttgactt gaggttagat tttttttata ttttgttttg tgttattttt
     4921 ttctttaaca tccctaaaat tttccttaca tgttttacta gccagatttt tcctcctctc
     4981 ctgactactc ccagtcatag ctgtccctct tctcttatgg agatccctcg acctgcagcc
     5041 caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     5101 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     5161 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     5221 gccagcggat ccgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     5281 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     5341 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     5401 cttttttgga ggcctaggct tttgcaaaaa gctaacttgt ttattgcagc ttataatggt
     5461 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     5521 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatccg ctgcattaat
     5581 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     5641 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     5701 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     5761 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     5821 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     5881 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     5941 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     6001 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     6061 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     6121 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     6181 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     6241 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     6301 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     6361 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6421 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     6481 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     6541 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     6601 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     6661 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     6721 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6781 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     6841 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     6901 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     6961 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     7021 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     7081 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     7141 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     7201 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     7261 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     7321 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     7381 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     7441 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     7501 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgggt
     7561 cgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
     7621 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     7681 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     7741 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     7801 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     7861 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     7921 ttagtcatcg ctattaccat ggtcgaggtg agccccacgt tctgcttcac tctccccatc
     7981 tcccccccct ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg
     8041 atgggggcgg gggggggggg ggggsgsggc gaggggcggg gcggggcgag gcggagaggt
     8101 gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg
     8161 cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc gcgctgcctt
     8221 cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc cccggctctg actgaccgcg
     8281 ttactcccac aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa ttagcgcttg
     8341 gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa gccttgaggg gctccgggag
     8401 ggccctttgt gcggggggag cggctcgggg ggtgcgtgcg tgtgtgtgtg cgtggggagc
     8461 gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct gcgggcgcgg cgcggggctt
     8521 tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg gcggtgcccc gcggtgcggg
     8581 gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt gcgtgggggg gtgagcaggg
     8641 ggtgtgggcg cgtcggtcgg gctgcaaccc cccctgcacc cccctccccg agttgctgag
     8701 cacggcccgg cttcgggtgc ggggctccgt acggggcgtg gcgcggggct cgccgtgccg
     8761 ggcggggggt ggcggcaggt gggggtgccg ggcggggcgg ggccgcctcg ggccggggag
     8821 ggctcggggg aggggcgcgg cggcccccgg agcgccggcg gctgtcgagg cgcggcgagc
     8881 cgcagccatt gccttttatg gtaatcgtgc gagagggcgc agggacttcc tttgtcccaa
     8941 atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc cctctagcgg gcgcggggcg
     9001 aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg gccttcgtgc gtcgccgcgc
     9061 cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc ggggggacgg ctgccttcgg
     9121 gggggacggg gcagggcggg gttcggcttc tggcgtgtga ccggcggctc tagagcctct
     9181 gctaaccatg ttcatgcctt cttctttttc ctacagctcc tgggcaacgt gctggttatt
     9241 gtgctgtctc atcattttgg caaagaattc ga
//
