LOCUS       sequence-202501-       11754 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SpCas9 (R1333S) fused to SaCas9 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11754)
  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 11754)
  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_SpCas9MT2-NLS-3xHA-NLS-SaCas9WT-2xNLS
FEATURES             Location/Qualifiers
     source          1..11754
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        217..235
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     217..234
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             285..332
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             351..371
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             378..4478
                     /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
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDSKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4497..4517
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4518..4607
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     CDS             4620..4646
                     /codon_start=1
                     /product="nuclear localization signal (Makkerh et al., 
                     1996)"
                     /label=NLS
                     /translation="PAAKKKKLD"
     CDS             4689..7844
                     /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             7851..7898
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7917..7943
                     /codon_start=1
                     /product="nuclear localization signal (Makkerh et al., 
                     1996)"
                     /label=NLS
                     /translation="PAAKKKKLD"
     polyA_signal    7985..8119
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8009..8028)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     8063..8082
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(8232..8252)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(8232..8250)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(8271..8287)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8271..8287)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8284..8306)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8295..8311
                     /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(8319..8325,8326..8343,8344..8349))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8364..8385
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8502..8519)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8673..9261)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8753..8772)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9432..10223,10224..10292))
                     /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     10055..10074
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10293..10397)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(10423..10878)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10560..10581)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     10772..10791
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(10952..11754,1..182)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 gggtacattg gcagtactcc cattgacgtc aatggcggta aatggcccgc gatggctgcc
       61 aagtacatcc ccattgacgt caatggggag gggcaatgac gcaaatgggc gttccattga
      121 cgtaaatggg cggtaggcgt gcctaatggg aggtctatat aagcaatgct cgtttaggga
      181 accgccattc tgcctgggga cgtcggagca agcttgattt aggtgacact atagaataca
      241 agctacttgt tctttttgca ggatcccatc gattcgaatt catgaaaagg ccggcggcca
      301 cgaaaaaggc cggccaggca aaaaagaaaa agctggacgg cagcggaagc cccaagaaga
      361 agaggaaggt accatgggac aagaagtact ccattgggct cgatatcggt accaacagcg
      421 tcggctgggc cgtcattacg gacgagtaca aggtgccgag caaaaaattc aaagttctgg
      481 gcaataccga tcgccacagc ataaagaaga acctcattgg agccctcctg ttcgactccg
      541 gggagacggc cgaagccacg cggctcaaaa gaacagcacg gcgcagatat acccgcagaa
      601 agaatcggat ctgctacctg caggagatct ttagtaatga gatggctaag gtggatgact
      661 ctttcttcca taggctggag gagtcctttt tggtggagga ggataaaaag cacgagcgcc
      721 acccaatctt tggcaatatc gtggacgagg tggcgtacca tgaaaagtac ccaaccatat
      781 atcatctgag gaagaagctg gtagacagta ctgataaggc tgacttgcgg ttgatctatc
      841 tcgcgctggc gcacatgatc aaatttcggg gacacttcct catcgagggg gacctgaacc
      901 cagacaacag cgatgtcgac aaactcttta tccaactggt tcagacttac aatcagcttt
      961 tcgaggagaa cccgatcaac gcatccggcg ttgacgccaa agcaatcctg agcgctaggc
     1021 tgtccaaatc ccggcggctc gaaaacctca tcgcacagct ccctggggag aagaagaacg
     1081 gcctgtttgg taatcttatc gccctgtcac tcgggctgac ccccaacttt aaatctaact
     1141 tcgacctggc cgaagatgcc aagctgcaac tgagcaaaga cacctacgat gatgatctcg
     1201 acaatctgct ggcccagatc ggcgaccagt acgcagacct ttttttggcg gcaaagaacc
     1261 tgtcagacgc cattctgctg agtgatattc tgcgagtgaa cacggagatc accaaagctc
     1321 cgctgagcgc tagtatgatc aagcgctatg atgagcacca ccaagacttg actttgctga
     1381 aggcccttgt cagacagcaa ctgcctgaga agtacaagga aattttcttc gatcagtcta
     1441 aaaatggcta cgccggatac attgacggcg gagcaagcca ggaggaattt tacaaattta
     1501 ttaagcccat cttggaaaaa atggacggca ccgaggagct gctggtaaag ctgaacagag
     1561 aagatctgtt gcgcaaacag cgcactttcg acaatggaag catcccccac cagattcacc
     1621 tgggcgaact gcacgctatc ctcaggcggc aagaggattt ctaccccttt ttgaaagata
     1681 acagggaaaa gattgagaaa atcctcacat ttcggatacc ctactatgta ggccccctcg
     1741 ctcggggaaa ttccagattc gcgtggatga ctcgcaaatc agaagagacc atcactccct
     1801 ggaacttcga ggaagtcgtg gataaggggg cctctgccca gtccttcatc gaaaggatga
     1861 ctaactttga taaaaatctg cctaacgaaa aggtgcttcc taaacactct ctgctgtacg
     1921 agtacttcac agtttataac gagctcacca aggtcaaata cgtcacagaa gggatgagaa
     1981 agccagcatt cctgtctgga gagcagaaga aagctatcgt ggacctcctc ttcaagacga
     2041 accggaaagt taccgtgaaa cagctcaaag aagactattt caaaaagatt gaatgtttcg
     2101 actctgttga aatcagcgga gtggaggatc gcttcaacgc atccctggga acgtatcacg
     2161 atctcctgaa aatcattaaa gacaaggact tcctggacaa tgaggagaac gaggacattc
     2221 ttgaggacat tgtcctcacc cttacgttgt ttgaagatag ggagatgatt gaagaacgct
     2281 tgaaaactta cgctcatctc ttcgacgaca aagtcatgaa acagctcaag agacgccgat
     2341 atacaggatg ggggcggctg tcaagaaaac tgatcaatgg catccgagac aagcagagtg
     2401 gaaagacaat cctggatttt cttaagtccg atggatttgc caaccggaac ttcatgcagt
     2461 tgatccatga tgactctctc acctttaagg aggacatcca gaaagcacaa gtttctggcc
     2521 agggggacag tcttcacgag cacatcgcta atcttgcagg tagcccagct atcaaaaagg
     2581 gaatactgca gaccgttaag gtcgtggatg aactcgtcaa agtaatggga aggcataagc
     2641 ccgagaatat cgttatcgag atggcccgag agaaccaaac tacccagaag ggacagaaga
     2701 acagtaggga aaggatgaag aggattgaag agggtataaa agaactgggg tcccaaatcc
     2761 ttaaggaaca cccagttgaa aacacccagc ttcagaatga gaagctctac ctgtactacc
     2821 tgcagaacgg cagggacatg tacgtggatc aggaactgga catcaaccgg ttgtccgact
     2881 acgacgtgga tcatatcgtg ccccaaagct ttctcaaaga tgattctatt gataataaag
     2941 tgttgacaag atccgataaa aatagaggga agagtgataa cgtcccctca gaagaagttg
     3001 tcaagaaaat gaaaaattat tggcggcagc tgctgaacgc caaactgatc acacaacgga
     3061 agttcgataa tctgactaag gctgaacgag gtggcctgtc tgagttggat aaagccggct
     3121 tcatcaaaag gcagcttgtt gagacacgcc agatcaccaa gcacgtggcc caaattctcg
     3181 attcacgcat gaacaccaag tacgatgaaa atgacaaact gattcgagag gtgaaagtta
     3241 ttactctgaa gtctaagctg gtctcagatt tcagaaagga ctttcagttt tataaggtga
     3301 gagagatcaa caattaccac catgcgcatg atgcctacct gaatgcagtg gtaggcactg
     3361 cacttatcaa aaaatatccc aagctggaat ctgaatttgt ttacggagac tataaagtgt
     3421 acgatgttag gaaaatgatc gcaaagtctg agcaggaaat aggcaaggcc accgctaagt
     3481 acttctttta cagcaatatt atgaattttt tcaagaccga gattacactg gccaatggag
     3541 agattcggaa gcgaccactt atcgaaacaa acggagaaac aggagaaatc gtgtgggaca
     3601 agggtaggga tttcgcgaca gtccgcaagg tcctgtccat gccgcaggtg aacatcgtta
     3661 aaaagaccga agtacagacc ggaggcttct ccaaggaaag tatcctcccg aaaaggaaca
     3721 gcgacaagct gatcgcacgc aaaaaagatt gggaccccaa gaaatacggc ggattcgatt
     3781 ctcctacagt cgcttacagt gtactggttg tggccaaagt ggagaaaggg aagtctaaaa
     3841 aactcaaaag cgtcaaggaa ctgctgggca tcacaatcat ggagcgatcc agcttcgaga
     3901 aaaaccccat cgactttctc gaagcgaaag gatataaaga ggtcaaaaaa gacctcatca
     3961 ttaagctgcc caagtactct ctctttgagc ttgaaaacgg ccggaaacga atgctcgcta
     4021 gtgcgggcga gctgcagaaa ggtaacgagc tggcactgcc ctctaaatac gttaatttct
     4081 tgtatctggc cagccactat gaaaagctca aagggtctcc cgaagataat gagcagaagc
     4141 agctgttcgt ggaacaacac aaacactacc ttgatgagat catcgagcaa ataagcgagt
     4201 tctccaaaag agtgatcctc gccgacgcta acctcgataa ggtgctttct gcttacaata
     4261 agcacaggga taagcccatc agggagcagg cagaaaacat tatccacttg tttactctga
     4321 ccaacttggg cgcgcctgca gccttcaagt acttcgacac caccatagac agcaagcggt
     4381 acacctctac aaaggaggtc ctggacgcca cactgattca tcagtcaatt acggggctct
     4441 atgaaacaag aatcgacctc tctcagctcg gtggagacgc tagcggcacc ggcgggccca
     4501 agaagaagag gaaggtatac ccatacgatg ttcctgacta tgcgggctat ccctatgacg
     4561 tcccggacta tgcaggatcg tatccttatg acgttccaga ttacgctgga tccgccgctc
     4621 cggcagctaa gaaaaagaaa ctggatttcg aatccggaac tagtggtatc cacggagtcc
     4681 cagcagccaa gcggaactac atcctgggcc tggacatcgg catcaccagc gtgggctacg
     4741 gcatcatcga ctacgagaca cgggacgtga tcgatgccgg cgtgcggctg ttcaaagagg
     4801 ccaacgtgga aaacaacgag ggcaggcgga gcaagagagg cgccagaagg ctgaagcggc
     4861 ggaggcggca tagaatccag agagtgaaga agctgctgtt cgactacaac ctgctgaccg
     4921 accacagcga gctgagcggc atcaacccct acgaggccag agtgaagggc ctgagccaga
     4981 agctgagcga ggaagagttc tctgccgccc tgctgcacct ggccaagaga agaggcgtgc
     5041 acaacgtgaa cgaggtggaa gaggacaccg gcaacgagct gtccaccaaa gagcagatca
     5101 gccggaacag caaggccctg gaagagaaat acgtggccga actgcagctg gaacggctga
     5161 agaaagacgg cgaagtgcgg ggcagcatca acagattcaa gaccagcgac tacgtgaaag
     5221 aagccaaaca gctgctgaag gtgcagaagg cctaccacca gctggaccag agcttcatcg
     5281 acacctacat cgacctgctg gaaacccggc ggacctacta tgagggacct ggcgagggca
     5341 gccccttcgg ctggaaggac atcaaagaat ggtacgagat gctgatgggc cactgcacct
     5401 acttccccga ggaactgcgg agcgtgaagt acgcctacaa cgccgacctg tacaacgccc
     5461 tgaacgacct gaacaatctc gtgatcacca gggacgagaa cgagaagctg gaatattacg
     5521 agaagttcca gatcatcgag aacgtgttca agcagaagaa gaagcccacc ctgaagcaga
     5581 tcgccaaaga aatcctcgtg aacgaagagg atattaaggg ctacagagtg accagcaccg
     5641 gcaagcccga gttcaccaac ctgaaggtgt accacgacat caaggacatt accgcccgga
     5701 aagagattat tgagaacgcc gagctgctgg atcagattgc caagatcctg accatctacc
     5761 agagcagcga ggacatccag gaagaactga ccaatctgaa ctccgagctg acccaggaag
     5821 agatcgagca gatctctaat ctgaagggct ataccggcac ccacaacctg agcctgaagg
     5881 ccatcaacct gatcctggac gagctgtggc acaccaacga caaccagatc gctatcttca
     5941 accggctgaa gctggtgccc aagaaggtgg acctgtccca gcagaaagag atccccacca
     6001 ccctggtgga cgacttcatc ctgagccccg tcgtgaagag aagcttcatc cagagcatca
     6061 aagtgatcaa cgccatcatc aagaagtacg gcctgcccaa cgacatcatt atcgagctgg
     6121 cccgcgagaa gaactccaag gacgcccaga aaatgatcaa cgagatgcag aagcggaacc
     6181 ggcagaccaa cgagcggatc gaggaaatca tccggaccac cggcaaagag aacgccaagt
     6241 acctgatcga gaagatcaag ctgcacgaca tgcaggaagg caagtgcctg tacagcctgg
     6301 aagccatccc tctggaagat ctgctgaaca accccttcaa ctatgaggtg gaccacatca
     6361 tccccagaag cgtgtccttc gacaacagct tcaacaacaa ggtgctcgtg aagcaggaag
     6421 aaaacagcaa gaagggcaac cggaccccat tccagtacct gagcagcagc gacagcaaga
     6481 tcagctacga aaccttcaag aagcacatcc tgaatctggc caagggcaag ggcagaatca
     6541 gcaagaccaa gaaagagtat ctgctggaag aacgggacat caacaggttc tccgtgcaga
     6601 aagacttcat caaccggaac ctggtggata ccagatacgc caccagaggc ctgatgaacc
     6661 tgctgcggag ctacttcaga gtgaacaacc tggacgtgaa agtgaagtcc atcaatggcg
     6721 gcttcaccag ctttctgcgg cggaagtgga agtttaagaa agagcggaac aaggggtaca
     6781 agcaccacgc cgaggacgcc ctgatcattg ccaacgccga tttcatcttc aaagagtgga
     6841 agaaactgga caaggccaaa aaagtgatgg aaaaccagat gttcgaggaa aagcaggccg
     6901 agagcatgcc cgagatcgaa accgagcagg agtacaaaga gatcttcatc accccccacc
     6961 agatcaagca cattaaggac ttcaaggact acaagtacag ccaccgggtg gacaagaagc
     7021 ctaatagaga gctgattaac gacaccctgt actccacccg gaaggacgac aagggcaaca
     7081 ccctgatcgt gaacaatctg aacggcctgt acgacaagga caatgacaag ctgaaaaagc
     7141 tgatcaacaa gagccccgaa aagctgctga tgtaccacca cgacccccag acctaccaga
     7201 aactgaagct gattatggaa cagtacggcg acgagaagaa tcccctgtac aagtactacg
     7261 aggaaaccgg gaactacctg accaagtact ccaaaaagga caacggcccc gtgatcaaga
     7321 agattaagta ttacggcaac aaactgaacg cccatctgga catcaccgac gactacccca
     7381 acagcagaaa caaggtcgtg aagctgtccc tgaagcccta cagattcgac gtgtacctgg
     7441 acaatggcgt gtacaagttc gtgaccgtga agaatctgga tgtgatcaaa aaagaaaact
     7501 actacgaagt gaatagcaag tgctatgagg aagctaagaa gctgaagaag atcagcaacc
     7561 aggccgagtt tatcgcctcc ttctacaaca acgatctgat caagatcaac ggcgagctgt
     7621 atagagtgat cggcgtgaac aacgacctgc tgaaccggat cgaagtgaac atgatcgaca
     7681 tcacctaccg cgagtacctg gaaaacatga acgacaagag gccccccagg atcattaaga
     7741 caatcgcctc caagacccag agcattaaga agtacagcac agacattctg ggcaacctgt
     7801 atgaagtgaa atctaagaag caccctcaga tcatcaaaaa gggccccggg aaaaggccgg
     7861 cggccacgaa aaaggccggc caggcaaaaa agaaaaagct ggacggcagc ggaagcccgg
     7921 cagctaagaa aaagaaactg gatctcgagt aataatctag aactatagtg agtcgtatta
     7981 cgtagatcca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     8041 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     8101 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     8161 ggaggtgtgg gaggtttttt aattcgcggc cgcggcgcca atgcattggg cccggtaccc
     8221 agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg
     8281 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     8341 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     8401 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     8461 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     8521 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     8581 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     8641 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     8701 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     8761 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     8821 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     8881 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     8941 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     9001 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     9061 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     9121 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     9181 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     9241 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     9301 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     9361 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     9421 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     9481 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     9541 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     9601 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     9661 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     9721 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     9781 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     9841 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     9901 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     9961 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
    10021 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
    10081 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
    10141 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
    10201 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
    10261 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
    10321 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
    10381 caaatagggg ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata
    10441 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
    10501 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
    10561 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
    10621 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
    10681 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
    10741 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
    10801 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
    10861 cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg caactgttgg gaagggcgat
    10921 cggtgcgggc ctcttcgcta ttacgccagt cgaccatagc caattcaata tggcgtatat
    10981 ggactcatgc caattcaata tggtggatct ggacctgtgc caattcaata tggcgtatat
    11041 ggactcgtgc caattcaata tggtggatct ggaccccagc caattcaata tggcggactt
    11101 ggcaccatgc caattcaata tggcggactt 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 ttggaaggac gcca
//
