LOCUS       sequence-204686-       11751 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SpCas9 fused to SaCas9 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11751)
  AUTHORS   Bolukbasi MF, Liu P, Luk K, Kwok SF, Gupta A, Amrani N, Sontheimer 
            EJ, Zhu LJ, Wolfe SA
  TITLE     Orthogonal Cas9-Cas9 chimeras provide a versatile platform for 
            genome editing.
  JOURNAL   Nat Commun. 2018 Nov 19;9(1):4856. doi: 10.1038/s41467-018-07310-x.
  PUBMED    30451839
REFERENCE   2  (bases 1 to 11751)
  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: "Nat 
            Commun."; date: "2018-11-19"; volume: "9(1):4856. doi"; pages: " 
            10.1038/s41467-018-07310-x"
COMMENT     Sequence Label: pCSD_2xNLS-SpCas9WT-NLS-3xHA-NLS-SaCas9WT-2xNLS
FEATURES             Location/Qualifiers
     source          1..11751
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        220..238
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     220..237
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             288..335
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             354..374
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             381..4481
                     /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
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             381..4481
                     /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
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             381..4481
                     /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
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4500..4520
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4521..4610
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     CDS             4623..4649
                     /codon_start=1
                     /product="nuclear localization signal (Makkerh et al., 
                     1996)"
                     /label=NLS
                     /translation="PAAKKKKLD"
     CDS             4692..7847
                     /codon_start=1
                     /product="Cas9 endonuclease from the Staphylococcus aureus 
                     Type II CRISPR/Cas system"
                     /label=SaCas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="KRNYILGLDIGITSVGYGIIDYETRDVIDAGVRLFKEANVENNEG
                     RRSKRGARRLKRRRRHRIQRVKKLLFDYNLLTDHSELSGINPYEARVKGLSQKLSEEEF
                     SAALLHLAKRRGVHNVNEVEEDTGNELSTKEQISRNSKALEEKYVAELQLERLKKDGEV
                     RGSINRFKTSDYVKEAKQLLKVQKAYHQLDQSFIDTYIDLLETRRTYYEGPGEGSPFGW
                     KDIKEWYEMLMGHCTYFPEELRSVKYAYNADLYNALNDLNNLVITRDENEKLEYYEKFQ
                     IIENVFKQKKKPTLKQIAKEILVNEEDIKGYRVTSTGKPEFTNLKVYHDIKDITARKEI
                     IENAELLDQIAKILTIYQSSEDIQEELTNLNSELTQEEIEQISNLKGYTGTHNLSLKAI
                     NLILDELWHTNDNQIAIFNRLKLVPKKVDLSQQKEIPTTLVDDFILSPVVKRSFIQSIK
                     VINAIIKKYGLPNDIIIELAREKNSKDAQKMINEMQKRNRQTNERIEEIIRTTGKENAK
                     YLIEKIKLHDMQEGKCLYSLEAIPLEDLLNNPFNYEVDHIIPRSVSFDNSFNNKVLVKQ
                     EENSKKGNRTPFQYLSSSDSKISYETFKKHILNLAKGKGRISKTKKEYLLEERDINRFS
                     VQKDFINRNLVDTRYATRGLMNLLRSYFRVNNLDVKVKSINGGFTSFLRRKWKFKKERN
                     KGYKHHAEDALIIANADFIFKEWKKLDKAKKVMENQMFEEKQAESMPEIETEQEYKEIF
                     ITPHQIKHIKDFKDYKYSHRVDKKPNRELINDTLYSTRKDDKGNTLIVNNLNGLYDKDN
                     DKLKKLINKSPEKLLMYHHDPQTYQKLKLIMEQYGDEKNPLYKYYEETGNYLTKYSKKD
                     NGPVIKKIKYYGNKLNAHLDITDDYPNSRNKVVKLSLKPYRFDVYLDNGVYKFVTVKNL
                     DVIKKENYYEVNSKCYEEAKKLKKISNQAEFIASFYNNDLIKINGELYRVIGVNNDLLN
                     RIEVNMIDITYREYLENMNDKRPPRIIKTIASKTQSIKKYSTDILGNLYEVKSKKHPQI
                     IKKG"
     CDS             7854..7901
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7920..7946
                     /codon_start=1
                     /product="nuclear localization signal (Makkerh et al., 
                     1996)"
                     /label=NLS
                     /translation="PAAKKKKLD"
     polyA_signal    7988..8122
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8012..8031)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     8066..8085
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(8235..8255)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(8235..8253)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(8274..8290)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8274..8290)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8287..8309)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8298..8314
                     /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(8322..8328,8329..8346,8347..8352))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8367..8388
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8505..8522)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8676..9264)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8756..8775)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9435..10226,10227..10295))
                     /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     10058..10077
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10296..10400)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(10426..10881)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10563..10584)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     10775..10794
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(10952..11751,1..185)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 ccagggtaca ttggcagtac tcccattgac gtcaatggcg gtaaatggcc cgcgatggct
       61 gccaagtaca tccccattga cgtcaatggg gaggggcaat gacgcaaatg ggcgttccat
      121 tgacgtaaat gggcggtagg cgtgcctaat gggaggtcta tataagcaat gctcgtttag
      181 ggaaccgcca ttctgcctgg ggacgtcgga gcaagcttga tttaggtgac actatagaat
      241 acaagctact tgttcttttt gcaggatccc atcgattcga attcatgaaa aggccggcgg
      301 ccacgaaaaa ggccggccag gcaaaaaaga aaaagctgga cggcagcgga agccccaaga
      361 agaagaggaa ggtaccatgg gacaagaagt actccattgg gctcgatatc ggtaccaaca
      421 gcgtcggctg ggccgtcatt acggacgagt acaaggtgcc gagcaaaaaa ttcaaagttc
      481 tgggcaatac cgatcgccac agcataaaga agaacctcat tggagccctc ctgttcgact
      541 ccggggagac ggccgaagcc acgcggctca aaagaacagc acggcgcaga tatacccgca
      601 gaaagaatcg gatctgctac ctgcaggaga tctttagtaa tgagatggct aaggtggatg
      661 actctttctt ccataggctg gaggagtcct ttttggtgga ggaggataaa aagcacgagc
      721 gccacccaat ctttggcaat atcgtggacg aggtggcgta ccatgaaaag tacccaacca
      781 tatatcatct gaggaagaag ctggtagaca gtactgataa ggctgacttg cggttgatct
      841 atctcgcgct ggcgcacatg atcaaatttc ggggacactt cctcatcgag ggggacctga
      901 acccagacaa cagcgatgtc gacaaactct ttatccaact ggttcagact tacaatcagc
      961 ttttcgagga gaacccgatc aacgcatccg gcgttgacgc caaagcaatc ctgagcgcta
     1021 ggctgtccaa atcccggcgg ctcgaaaacc tcatcgcaca gctccctggg gagaagaaga
     1081 acggcctgtt tggtaatctt atcgccctgt cactcgggct gacccccaac tttaaatcta
     1141 acttcgacct ggccgaagat gccaagctgc aactgagcaa agacacctac gatgatgatc
     1201 tcgacaatct gctggcccag atcggcgacc agtacgcaga cctttttttg gcggcaaaga
     1261 acctgtcaga cgccattctg ctgagtgata ttctgcgagt gaacacggag atcaccaaag
     1321 ctccgctgag cgctagtatg atcaagcgct atgatgagca ccaccaagac ttgactttgc
     1381 tgaaggccct tgtcagacag caactgcctg agaagtacaa ggaaattttc ttcgatcagt
     1441 ctaaaaatgg ctacgccgga tacattgacg gcggagcaag ccaggaggaa ttttacaaat
     1501 ttattaagcc catcttggaa aaaatggacg gcaccgagga gctgctggta aagctgaaca
     1561 gagaagatct gttgcgcaaa cagcgcactt tcgacaatgg aagcatcccc caccagattc
     1621 acctgggcga actgcacgct atcctcaggc ggcaagagga tttctacccc tttttgaaag
     1681 ataacaggga aaagattgag aaaatcctca catttcggat accctactat gtaggccccc
     1741 tcgctcgggg aaattccaga ttcgcgtgga tgactcgcaa atcagaagag accatcactc
     1801 cctggaactt cgaggaagtc gtggataagg gggcctctgc ccagtccttc atcgaaagga
     1861 tgactaactt tgataaaaat ctgcctaacg aaaaggtgct tcctaaacac tctctgctgt
     1921 acgagtactt cacagtttat aacgagctca ccaaggtcaa atacgtcaca gaagggatga
     1981 gaaagccagc attcctgtct ggagagcaga agaaagctat cgtggacctc ctcttcaaga
     2041 cgaaccggaa agttaccgtg aaacagctca aagaagacta tttcaaaaag attgaatgtt
     2101 tcgactctgt tgaaatcagc ggagtggagg atcgcttcaa cgcatccctg ggaacgtatc
     2161 acgatctcct gaaaatcatt aaagacaagg acttcctgga caatgaggag aacgaggaca
     2221 ttcttgagga cattgtcctc acccttacgt tgtttgaaga tagggagatg attgaagaac
     2281 gcttgaaaac ttacgctcat ctcttcgacg acaaagtcat gaaacagctc aagagacgcc
     2341 gatatacagg atgggggcgg ctgtcaagaa aactgatcaa tggcatccga gacaagcaga
     2401 gtggaaagac aatcctggat tttcttaagt ccgatggatt tgccaaccgg aacttcatgc
     2461 agttgatcca tgatgactct ctcaccttta aggaggacat ccagaaagca caagtttctg
     2521 gccaggggga cagtcttcac gagcacatcg ctaatcttgc aggtagccca gctatcaaaa
     2581 agggaatact gcagaccgtt aaggtcgtgg atgaactcgt caaagtaatg ggaaggcata
     2641 agcccgagaa tatcgttatc gagatggccc gagagaacca aactacccag aagggacaga
     2701 agaacagtag ggaaaggatg aagaggattg aagagggtat aaaagaactg gggtcccaaa
     2761 tccttaagga acacccagtt gaaaacaccc agcttcagaa tgagaagctc tacctgtact
     2821 acctgcagaa cggcagggac atgtacgtgg atcaggaact ggacatcaac cggttgtccg
     2881 actacgacgt ggatcatatc gtgccccaaa gctttctcaa agatgattct attgataata
     2941 aagtgttgac aagatccgat aaaaatagag ggaagagtga taacgtcccc tcagaagaag
     3001 ttgtcaagaa aatgaaaaat tattggcggc agctgctgaa cgccaaactg atcacacaac
     3061 ggaagttcga taatctgact aaggctgaac gaggtggcct gtctgagttg gataaagccg
     3121 gcttcatcaa aaggcagctt gttgagacac gccagatcac caagcacgtg gcccaaattc
     3181 tcgattcacg catgaacacc aagtacgatg aaaatgacaa actgattcga gaggtgaaag
     3241 ttattactct gaagtctaag ctggtctcag atttcagaaa ggactttcag ttttataagg
     3301 tgagagagat caacaattac caccatgcgc atgatgccta cctgaatgca gtggtaggca
     3361 ctgcacttat caaaaaatat cccaagctgg aatctgaatt tgtttacgga gactataaag
     3421 tgtacgatgt taggaaaatg atcgcaaagt ctgagcagga aataggcaag gccaccgcta
     3481 agtacttctt ttacagcaat attatgaatt ttttcaagac cgagattaca ctggccaatg
     3541 gagagattcg gaagcgacca cttatcgaaa caaacggaga aacaggagaa atcgtgtggg
     3601 acaagggtag ggatttcgcg acagtccgca aggtcctgtc catgccgcag gtgaacatcg
     3661 ttaaaaagac cgaagtacag accggaggct tctccaagga aagtatcctc ccgaaaagga
     3721 acagcgacaa gctgatcgca cgcaaaaaag attgggaccc caagaaatac ggcggattcg
     3781 attctcctac agtcgcttac agtgtactgg ttgtggccaa agtggagaaa gggaagtcta
     3841 aaaaactcaa aagcgtcaag gaactgctgg gcatcacaat catggagcga tccagcttcg
     3901 agaaaaaccc catcgacttt ctcgaagcga aaggatataa agaggtcaaa aaagacctca
     3961 tcattaagct gcccaagtac tctctctttg agcttgaaaa cggccggaaa cgaatgctcg
     4021 ctagtgcggg cgagctgcag aaaggtaacg agctggcact gccctctaaa tacgttaatt
     4081 tcttgtatct ggccagccac tatgaaaagc tcaaagggtc tcccgaagat aatgagcaga
     4141 agcagctgtt cgtggaacaa cacaaacact accttgatga gatcatcgag caaataagcg
     4201 agttctccaa aagagtgatc ctcgccgacg ctaacctcga taaggtgctt tctgcttaca
     4261 ataagcacag ggataagccc atcagggagc aggcagaaaa cattatccac ttgtttactc
     4321 tgaccaactt gggcgcgcct gcagccttca agtacttcga caccaccata gacagaaagc
     4381 ggtacacctc tacaaaggag gtcctggacg ccacactgat tcatcagtca attacggggc
     4441 tctatgaaac aagaatcgac ctctctcagc tcggtggaga cgctagcggc accggcgggc
     4501 ccaagaagaa gaggaaggta tacccatacg atgttcctga ctatgcgggc tatccctatg
     4561 acgtcccgga ctatgcagga tcgtatcctt atgacgttcc agattacgct ggatccgccg
     4621 ctccggcagc taagaaaaag aaactggatt tcgaatccgg aactagtggt atccacggag
     4681 tcccagcagc caagcggaac tacatcctgg gcctggacat cggcatcacc agcgtgggct
     4741 acggcatcat cgactacgag acacgggacg tgatcgatgc cggcgtgcgg ctgttcaaag
     4801 aggccaacgt ggaaaacaac gagggcaggc ggagcaagag aggcgccaga aggctgaagc
     4861 ggcggaggcg gcatagaatc cagagagtga agaagctgct gttcgactac aacctgctga
     4921 ccgaccacag cgagctgagc ggcatcaacc cctacgaggc cagagtgaag ggcctgagcc
     4981 agaagctgag cgaggaagag ttctctgccg ccctgctgca cctggccaag agaagaggcg
     5041 tgcacaacgt gaacgaggtg gaagaggaca ccggcaacga gctgtccacc aaagagcaga
     5101 tcagccggaa cagcaaggcc ctggaagaga aatacgtggc cgaactgcag ctggaacggc
     5161 tgaagaaaga cggcgaagtg cggggcagca tcaacagatt caagaccagc gactacgtga
     5221 aagaagccaa acagctgctg aaggtgcaga aggcctacca ccagctggac cagagcttca
     5281 tcgacaccta catcgacctg ctggaaaccc ggcggaccta ctatgaggga cctggcgagg
     5341 gcagcccctt cggctggaag gacatcaaag aatggtacga gatgctgatg ggccactgca
     5401 cctacttccc cgaggaactg cggagcgtga agtacgccta caacgccgac ctgtacaacg
     5461 ccctgaacga cctgaacaat ctcgtgatca ccagggacga gaacgagaag ctggaatatt
     5521 acgagaagtt ccagatcatc gagaacgtgt tcaagcagaa gaagaagccc accctgaagc
     5581 agatcgccaa agaaatcctc gtgaacgaag aggatattaa gggctacaga gtgaccagca
     5641 ccggcaagcc cgagttcacc aacctgaagg tgtaccacga catcaaggac attaccgccc
     5701 ggaaagagat tattgagaac gccgagctgc tggatcagat tgccaagatc ctgaccatct
     5761 accagagcag cgaggacatc caggaagaac tgaccaatct gaactccgag ctgacccagg
     5821 aagagatcga gcagatctct aatctgaagg gctataccgg cacccacaac ctgagcctga
     5881 aggccatcaa cctgatcctg gacgagctgt ggcacaccaa cgacaaccag atcgctatct
     5941 tcaaccggct gaagctggtg cccaagaagg tggacctgtc ccagcagaaa gagatcccca
     6001 ccaccctggt ggacgacttc atcctgagcc ccgtcgtgaa gagaagcttc atccagagca
     6061 tcaaagtgat caacgccatc atcaagaagt acggcctgcc caacgacatc attatcgagc
     6121 tggcccgcga gaagaactcc aaggacgccc agaaaatgat caacgagatg cagaagcgga
     6181 accggcagac caacgagcgg atcgaggaaa tcatccggac caccggcaaa gagaacgcca
     6241 agtacctgat cgagaagatc aagctgcacg acatgcagga aggcaagtgc ctgtacagcc
     6301 tggaagccat ccctctggaa gatctgctga acaacccctt caactatgag gtggaccaca
     6361 tcatccccag aagcgtgtcc ttcgacaaca gcttcaacaa caaggtgctc gtgaagcagg
     6421 aagaaaacag caagaagggc aaccggaccc cattccagta cctgagcagc agcgacagca
     6481 agatcagcta cgaaaccttc aagaagcaca tcctgaatct ggccaagggc aagggcagaa
     6541 tcagcaagac caagaaagag tatctgctgg aagaacggga catcaacagg ttctccgtgc
     6601 agaaagactt catcaaccgg aacctggtgg ataccagata cgccaccaga ggcctgatga
     6661 acctgctgcg gagctacttc agagtgaaca acctggacgt gaaagtgaag tccatcaatg
     6721 gcggcttcac cagctttctg cggcggaagt ggaagtttaa gaaagagcgg aacaaggggt
     6781 acaagcacca cgccgaggac gccctgatca ttgccaacgc cgatttcatc ttcaaagagt
     6841 ggaagaaact ggacaaggcc aaaaaagtga tggaaaacca gatgttcgag gaaaagcagg
     6901 ccgagagcat gcccgagatc gaaaccgagc aggagtacaa agagatcttc atcacccccc
     6961 accagatcaa gcacattaag gacttcaagg actacaagta cagccaccgg gtggacaaga
     7021 agcctaatag agagctgatt aacgacaccc tgtactccac ccggaaggac gacaagggca
     7081 acaccctgat cgtgaacaat ctgaacggcc tgtacgacaa ggacaatgac aagctgaaaa
     7141 agctgatcaa caagagcccc gaaaagctgc tgatgtacca ccacgacccc cagacctacc
     7201 agaaactgaa gctgattatg gaacagtacg gcgacgagaa gaatcccctg tacaagtact
     7261 acgaggaaac cgggaactac ctgaccaagt actccaaaaa ggacaacggc cccgtgatca
     7321 agaagattaa gtattacggc aacaaactga acgcccatct ggacatcacc gacgactacc
     7381 ccaacagcag aaacaaggtc gtgaagctgt ccctgaagcc ctacagattc gacgtgtacc
     7441 tggacaatgg cgtgtacaag ttcgtgaccg tgaagaatct ggatgtgatc aaaaaagaaa
     7501 actactacga agtgaatagc aagtgctatg aggaagctaa gaagctgaag aagatcagca
     7561 accaggccga gtttatcgcc tccttctaca acaacgatct gatcaagatc aacggcgagc
     7621 tgtatagagt gatcggcgtg aacaacgacc tgctgaaccg gatcgaagtg aacatgatcg
     7681 acatcaccta ccgcgagtac ctggaaaaca tgaacgacaa gaggcccccc aggatcatta
     7741 agacaatcgc ctccaagacc cagagcatta agaagtacag cacagacatt ctgggcaacc
     7801 tgtatgaagt gaaatctaag aagcaccctc agatcatcaa aaagggcccc gggaaaaggc
     7861 cggcggccac gaaaaaggcc ggccaggcaa aaaagaaaaa gctggacggc agcggaagcc
     7921 cggcagctaa gaaaaagaaa ctggatctcg agtaataatc tagaactata gtgagtcgta
     7981 ttacgtagat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     8041 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     8101 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     8161 gggggaggtg tgggaggttt tttaattcgc ggccgcggcg ccaatgcatt gggcccggta
     8221 cccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag
     8281 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccgggagc
     8341 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     8401 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     8461 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     8521 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     8581 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     8641 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     8701 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     8761 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     8821 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     8881 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     8941 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     9001 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     9061 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     9121 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     9181 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     9241 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     9301 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     9361 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     9421 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     9481 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     9541 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     9601 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     9661 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     9721 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     9781 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     9841 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     9901 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     9961 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
    10021 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
    10081 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
    10141 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
    10201 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
    10261 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
    10321 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
    10381 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta
    10441 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
    10501 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
    10561 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
    10621 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
    10681 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
    10741 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
    10801 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
    10861 atgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt tgggaagggc
    10921 gatcggtgcg ggcctcttcg ctattacgcc agtcgaccgc caattcaata tggcgtatat
    10981 ggactcatgc caattcaata tggtggatct ggacctgtgc caattcaata tggcgtatat
    11041 ggactcgtgc caattcaata tggtggatct ggaccccagc caattcaata tggcggactt
    11101 ggcaccatgc caattcaata tggcggacct ggcactgtgc caactgggga ggggtctact
    11161 tggcacggtg ccaagtttga ggaggggtct tggccctgtg ccaagtccgc catattgaat
    11221 tggcatggtg ccaataatgg cggccatatt ggctatatgc caggatcaat atataggcaa
    11281 tatccaatat ggccctatgc caatatggct attggccagg ttcaatacta tgtattggcc
    11341 ctatgccata tagtattcca tatatgggtt ttcctattga cgtagatagc ccctcccaat
    11401 gggcggtccc atataccata tatggggctt cctaataccg cccatagcca ctcccccatt
    11461 gacgtcaatg gtctctatat atggtctttc ctattgacgt catatgggcg gtcctattga
    11521 cgtatatggc gcctccccca ttgacgtcaa ttacggtaaa tggcccgcct ggctcaatgc
    11581 ccattgacgt caataggacc acccaccatt gacgtcaatg ggatggctca ttgcccattc
    11641 atatccgttc tcacgccccc tattgacgtc aatgacggta aatggcccac ttggcagtac
    11701 atcaatatct attaatagta acttggcaag tacattacta ttggaagtac g
//
