LOCUS       sequence-204774-       11834 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SpCas9 (R1333S) fused to NmCas9 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11834)
  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 11834)
  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-SpCas9MT2-NLS-3xHA-2XNLS-NmCas9WT-2...
FEATURES             Location/Qualifiers
     source          1..11834
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        38..56
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     38..55
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             106..153
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             172..192
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             199..4299
                     /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             4312..4332
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4333..4422
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     CDS             4435..4461
                     /codon_start=1
                     /product="nuclear localization signal (Makkerh et al., 
                     1996)"
                     /label=NLS
                     /translation="PAAKKKKLD"
     CDS             4477..4497
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4498..7740
                     /codon_start=1
                     /product="Cas9 endonuclease from the Neisseria meningitidis
                     Type II CRISPR/Cas system (Esvelt et al., 2013)"
                     /label=NmCas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="AAFKPNPINYILGLDIGIASVGWAMVEIDEDENPICLIDLGVRVF
                     ERAEVPKTGDSLAMARRLARSVRRLTRRRAHRLLRARRLLKREGVLQAADFDENGLIKS
                     LPNTPWQLRAAALDRKLTPLEWSAVLLHLIKHRGYLSQRKNEGETADKELGALLKGVAD
                     NAHALQTGDFRTPAELALNKFEKESGHIRNQRGDYSHTFSRKDLQAELILLFEKQKEFG
                     NPHVSGGLKEGIETLLMTQRPALSGDAVQKMLGHCTFEPAEPKAAKNTYTAERFIWLTK
                     LNNLRILEQGSERPLTDTERATLMDEPYRKSKLTYAQARKLLGLEDTAFFKGLRYGKDN
                     AEASTLMEMKAYHAISRALEKEGLKDKKSPLNLSPELQDEIGTAFSLFKTDEDITGRLK
                     DRIQPEILEALLKHISFDKFVQISLKALRRIVPLMEQGKRYDEACAEIYGDHYGKKNTE
                     EKIYLPPIPADEIRNPVVLRALSQARKVINGVVRRYGSPARIHIETAREVGKSFKDRKE
                     IEKRQEENRKDREKAAAKFREYFPNFVGEPKSKDILKLRLYEQQHGKCLYSGKEINLGR
                     LNEKGYVEIDHALPFSRTWDDSFNNKVLVLGSENQNKGNQTPYEYFNGKDNSREWQEFK
                     ARVETSRFPRSKKQRILLQKFDEDGFKERNLNDTRYVNRFLCQFVADRMRLTGKGKKRV
                     FASNGQITNLLRGFWGLRKVRAENDRHHALDAVVVACSTVAMQQKITRFVRYKEMNAFD
                     GKTIDKETGEVLHQKTHFPQPWEFFAQEVMIRVFGKPDGKPEFEEADTPEKLRTLLAEK
                     LSSRPEAVHEYVTPLFVSRAPNRKMSGQGHMETVKSAKRLDEGVSVLRVPLTQLKLKDL
                     EKMVNREREPKLYEALKARLEAHKDDPAKAFAEPFYKYDKAGNRTQQVKAVRVEQVQKT
                     GVWVRNHNGIADNATMVRVDVFEKGDKYYLVPIYSWQVAKGILPDRAVVQGKDEEDWQL
                     IDDSFNFKFSLHPNDLVEVITKKARMFGYFASCHRGTGNINIRIHDLDHKIGKNGILEG
                     IGVKTALSFQKYQIDELGKEIRPCRLKKRPPVR"
     regulatory      7327..7336
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7753..7773
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7789..7836
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     polyA_signal    7889..8023
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7913..7932)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7967..7986
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(8136..8156)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(8136..8154)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(8175..8191)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8175..8191)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8188..8210)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8199..8215
                     /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(8223..8229,8230..8247,8248..8253))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8268..8289
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8406..8423)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8577..9165)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8657..8676)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9336..10127,10128..10196))
                     /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     9959..9978
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10197..10301)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(10327..10782)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10464..10485)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     10676..10695
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(10853..11834,1..3)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 aaccgccatt ctgcctgggg acgtcggagc aagcttgatt taggtgacac tatagaatac
       61 aagctacttg ttctttttgc aggatcccat cgattcgaat tcatgaaaag gccggcggcc
      121 acgaaaaagg ccggccaggc aaaaaagaaa aagctggacg gcagcggaag ccccaagaag
      181 aagaggaagg taccatggga caagaagtac tccattgggc tcgatatcgg taccaacagc
      241 gtcggctggg ccgtcattac ggacgagtac aaggtgccga gcaaaaaatt caaagttctg
      301 ggcaataccg atcgccacag cataaagaag aacctcattg gagccctcct gttcgactcc
      361 ggggagacgg ccgaagccac gcggctcaaa agaacagcac ggcgcagata tacccgcaga
      421 aagaatcgga tctgctacct gcaggagatc tttagtaatg agatggctaa ggtggatgac
      481 tctttcttcc ataggctgga ggagtccttt ttggtggagg aggataaaaa gcacgagcgc
      541 cacccaatct ttggcaatat cgtggacgag gtggcgtacc atgaaaagta cccaaccata
      601 tatcatctga ggaagaagct ggtagacagt actgataagg ctgacttgcg gttgatctat
      661 ctcgcgctgg cgcacatgat caaatttcgg ggacacttcc tcatcgaggg ggacctgaac
      721 ccagacaaca gcgatgtcga caaactcttt atccaactgg ttcagactta caatcagctt
      781 ttcgaggaga acccgatcaa cgcatccggc gttgacgcca aagcaatcct gagcgctagg
      841 ctgtccaaat cccggcggct cgaaaacctc atcgcacagc tccctgggga gaagaagaac
      901 ggcctgtttg gtaatcttat cgccctgtca ctcgggctga cccccaactt taaatctaac
      961 ttcgacctgg ccgaagatgc caagctgcaa ctgagcaaag acacctacga tgatgatctc
     1021 gacaatctgc tggcccagat cggcgaccag tacgcagacc tttttttggc ggcaaagaac
     1081 ctgtcagacg ccattctgct gagtgatatt ctgcgagtga acacggagat caccaaagct
     1141 ccgctgagcg ctagtatgat caagcgctat gatgagcacc accaagactt gactttgctg
     1201 aaggcccttg tcagacagca actgcctgag aagtacaagg aaattttctt cgatcagtct
     1261 aaaaatggct acgccggata cattgacggc ggagcaagcc aggaggaatt ttacaaattt
     1321 attaagccca tcttggaaaa aatggacggc accgaggagc tgctggtaaa gctgaacaga
     1381 gaagatctgt tgcgcaaaca gcgcactttc gacaatggaa gcatccccca ccagattcac
     1441 ctgggcgaac tgcacgctat cctcaggcgg caagaggatt tctacccctt tttgaaagat
     1501 aacagggaaa agattgagaa aatcctcaca tttcggatac cctactatgt aggccccctc
     1561 gctcggggaa attccagatt cgcgtggatg actcgcaaat cagaagagac catcactccc
     1621 tggaacttcg aggaagtcgt ggataagggg gcctctgccc agtccttcat cgaaaggatg
     1681 actaactttg ataaaaatct gcctaacgaa aaggtgcttc ctaaacactc tctgctgtac
     1741 gagtacttca cagtttataa cgagctcacc aaggtcaaat acgtcacaga agggatgaga
     1801 aagccagcat tcctgtctgg agagcagaag aaagctatcg tggacctcct cttcaagacg
     1861 aaccggaaag ttaccgtgaa acagctcaaa gaagactatt tcaaaaagat tgaatgtttc
     1921 gactctgttg aaatcagcgg agtggaggat cgcttcaacg catccctggg aacgtatcac
     1981 gatctcctga aaatcattaa agacaaggac ttcctggaca atgaggagaa cgaggacatt
     2041 cttgaggaca ttgtcctcac ccttacgttg tttgaagata gggagatgat tgaagaacgc
     2101 ttgaaaactt acgctcatct cttcgacgac aaagtcatga aacagctcaa gagacgccga
     2161 tatacaggat gggggcggct gtcaagaaaa ctgatcaatg gcatccgaga caagcagagt
     2221 ggaaagacaa tcctggattt tcttaagtcc gatggatttg ccaaccggaa cttcatgcag
     2281 ttgatccatg atgactctct cacctttaag gaggacatcc agaaagcaca agtttctggc
     2341 cagggggaca gtcttcacga gcacatcgct aatcttgcag gtagcccagc tatcaaaaag
     2401 ggaatactgc agaccgttaa ggtcgtggat gaactcgtca aagtaatggg aaggcataag
     2461 cccgagaata tcgttatcga gatggcccga gagaaccaaa ctacccagaa gggacagaag
     2521 aacagtaggg aaaggatgaa gaggattgaa gagggtataa aagaactggg gtcccaaatc
     2581 cttaaggaac acccagttga aaacacccag cttcagaatg agaagctcta cctgtactac
     2641 ctgcagaacg gcagggacat gtacgtggat caggaactgg acatcaaccg gttgtccgac
     2701 tacgacgtgg atcatatcgt gccccaaagc tttctcaaag atgattctat tgataataaa
     2761 gtgttgacaa gatccgataa aaatagaggg aagagtgata acgtcccctc agaagaagtt
     2821 gtcaagaaaa tgaaaaatta ttggcggcag ctgctgaacg ccaaactgat cacacaacgg
     2881 aagttcgata atctgactaa ggctgaacga ggtggcctgt ctgagttgga taaagccggc
     2941 ttcatcaaaa ggcagcttgt tgagacacgc cagatcacca agcacgtggc ccaaattctc
     3001 gattcacgca tgaacaccaa gtacgatgaa aatgacaaac tgattcgaga ggtgaaagtt
     3061 attactctga agtctaagct ggtctcagat ttcagaaagg actttcagtt ttataaggtg
     3121 agagagatca acaattacca ccatgcgcat gatgcctacc tgaatgcagt ggtaggcact
     3181 gcacttatca aaaaatatcc caagctggaa tctgaatttg tttacggaga ctataaagtg
     3241 tacgatgtta ggaaaatgat cgcaaagtct gagcaggaaa taggcaaggc caccgctaag
     3301 tacttctttt acagcaatat tatgaatttt ttcaagaccg agattacact ggccaatgga
     3361 gagattcgga agcgaccact tatcgaaaca aacggagaaa caggagaaat cgtgtgggac
     3421 aagggtaggg atttcgcgac agtccgcaag gtcctgtcca tgccgcaggt gaacatcgtt
     3481 aaaaagaccg aagtacagac cggaggcttc tccaaggaaa gtatcctccc gaaaaggaac
     3541 agcgacaagc tgatcgcacg caaaaaagat tgggacccca agaaatacgg cggattcgat
     3601 tctcctacag tcgcttacag tgtactggtt gtggccaaag tggagaaagg gaagtctaaa
     3661 aaactcaaaa gcgtcaagga actgctgggc atcacaatca tggagcgatc cagcttcgag
     3721 aaaaacccca tcgactttct cgaagcgaaa ggatataaag aggtcaaaaa agacctcatc
     3781 attaagctgc ccaagtactc tctctttgag cttgaaaacg gccggaaacg aatgctcgct
     3841 agtgcgggcg agctgcagaa aggtaacgag ctggcactgc cctctaaata cgttaatttc
     3901 ttgtatctgg ccagccacta tgaaaagctc aaagggtctc ccgaagataa tgagcagaag
     3961 cagctgttcg tggaacaaca caaacactac cttgatgaga tcatcgagca aataagcgag
     4021 ttctccaaaa gagtgatcct cgccgacgct aacctcgata aggtgctttc tgcttacaat
     4081 aagcacaggg ataagcccat cagggagcag gcagaaaaca ttatccactt gtttactctg
     4141 accaacttgg gcgcgcctgc agccttcaag tacttcgaca ccaccataga cagcaagcgg
     4201 tacacctcta caaaggaggt cctggacgcc acactgattc atcagtcaat tacggggctc
     4261 tatgaaacaa gaatcgacct ctctcagctc ggtggagacg gcaccggcgg gcccaagaag
     4321 aagaggaagg tatacccata cgatgttcct gactatgcgg gctatcccta tgacgtcccg
     4381 gactatgcag gatcgtatcc ttatgacgtt ccagattacg ctggatccgc cgctccggca
     4441 gctaagaaaa agaaactgga tttcgaatcc ggaatgccca agaagaagag gaaggtggcc
     4501 gccttcaagc ccaaccccat caactacatc ctgggcctgg acatcggcat cgccagcgtg
     4561 ggctgggcca tggtggagat cgacgaggac gagaacccca tctgcctgat cgacctgggt
     4621 gtgcgcgtgt tcgagcgcgc tgaggtgccc aagactggtg acagtctggc tatggctcgc
     4681 cggcttgctc gctctgttcg gcgccttact cgccggcgcg ctcaccgcct tctgcgcgct
     4741 cgccgcctgc tgaagcgcga gggtgtgctg caggctgccg acttcgacga gaacggcctg
     4801 atcaagagcc tgcccaacac tccttggcag ctgcgcgctg ccgctctgga ccgcaagctg
     4861 actcctctgg agtggagcgc cgtgctgctg cacctgatca agcaccgcgg ctacctgagc
     4921 cagcgcaaga acgagggcga gaccgccgac aaggagctgg gtgctctgct gaagggcgtg
     4981 gccgacaacg cccacgccct gcagactggt gacttccgca ctcctgctga gctggccctg
     5041 aacaagttcg agaaggagag cggccacatc cgcaaccagc gcggcgacta cagccacacc
     5101 ttcagccgca aggacctgca ggccgagctg atcctgctgt tcgagaagca gaaggagttc
     5161 ggcaaccccc acgtgagcgg cggcctgaag gagggcatcg agaccctgct gatgacccag
     5221 cgccccgccc tgagcggcga cgccgtgcag aagatgctgg gccactgcac cttcgagcca
     5281 gccgagccca aggccgccaa gaacacctac accgccgagc gcttcatctg gctgaccaag
     5341 ctgaacaacc tgcgcatcct ggagcagggc agcgagcgcc ccctgaccga caccgagcgc
     5401 gccaccctga tggacgagcc ctaccgcaag agcaagctga cctacgccca ggcccgcaag
     5461 ctgctgggtc tggaggacac cgccttcttc aagggcctgc gctacggcaa ggacaacgcc
     5521 gaggccagca ccctgatgga gatgaaggcc taccacgcca tcagccgcgc cctggagaag
     5581 gagggcctga aggacaagaa gagtcctctg aacctgagcc ccgagctgca ggacgagatc
     5641 ggcaccgcct tcagcctgtt caagaccgac gaggacatca ccggccgcct gaaggaccgc
     5701 atccagcccg agatcctgga ggccctgctg aagcacatca gcttcgacaa gttcgtgcag
     5761 atcagcctga aggccctgcg ccgcatcgtg cccctgatgg agcagggcaa gcgctacgac
     5821 gaggcctgcg ccgagatcta cggcgaccac tacggcaaga agaacaccga ggagaagatc
     5881 tacctgcctc ctatccccgc cgacgagatc cgcaaccccg tggtgctgcg cgccctgagc
     5941 caggcccgca aggtgatcaa cggcgtggtg cgccgctacg gcagccccgc ccgcatccac
     6001 atcgagaccg cccgcgaggt gggcaagagc ttcaaggacc gcaaggagat cgagaagcgc
     6061 caggaggaga accgcaagga ccgcgagaag gccgccgcca agttccgcga gtacttcccc
     6121 aacttcgtgg gcgagcccaa gagcaaggac atcctgaagc tgcgcctgta cgagcagcag
     6181 cacggcaagt gcctgtacag cggcaaggag atcaacctgg gccgcctgaa cgagaagggc
     6241 tacgtggaga tcgaccatgc cctgcccttc agccgcacct gggacgacag cttcaacaac
     6301 aaggtgctgg tgctgggcag cgagaatcag aacaagggca accagacccc ctacgagtac
     6361 ttcaacggca aggacaacag ccgcgagtgg caggagttca aggcccgcgt ggagaccagc
     6421 cgcttccccc gcagcaagaa gcagcgcatc ctgctgcaga agttcgacga ggacggcttc
     6481 aaggagcgca acctgaacga cacccgctac gtgaaccgct tcctgtgcca gttcgtggcc
     6541 gaccgcatgc gcctgaccgg caagggcaag aagcgcgtgt tcgccagcaa cggccagatc
     6601 accaacctgc tgcgcggctt ctggggcctg cgcaaggtgc gcgccgagaa cgaccgccac
     6661 cacgccctgg acgccgtggt ggtggcctgc agcaccgtgg ccatgcagca gaagatcacc
     6721 cgcttcgtgc gctacaagga gatgaacgcc ttcgacggta aaaccatcga caaggagacc
     6781 ggcgaggtgc tgcaccagaa gacccacttc ccccagccct gggagttctt cgcccaggag
     6841 gtgatgatcc gcgtgttcgg caagcccgac ggcaagcccg agttcgagga ggccgacacc
     6901 cccgagaagc tgcgcaccct gctggccgag aagctgagca gccgccctga ggccgtgcac
     6961 gagtacgtga ctcctctgtt cgtgagccgc gcccccaacc gcaagatgag cggtcagggt
     7021 cacatggaga ccgtgaagag cgccaagcgc ctggacgagg gcgtgagcgt gctgcgcgtg
     7081 cccctgaccc agctgaagct gaaggacctg gagaagatgg tgaaccgcga gcgcgagccc
     7141 aagctgtacg aggccctgaa ggcccgcctg gaggcccaca aggacgaccc cgccaaggcc
     7201 ttcgccgagc ccttctacaa gtacgacaag gccggcaacc gcacccagca ggtgaaggcc
     7261 gtgcgcgtgg agcaggtgca gaagaccggc gtgtgggtgc gcaaccacaa cggcatcgcc
     7321 gacaacgcca ccatggtgcg cgtggacgtg ttcgagaagg gcgacaagta ctacctggtg
     7381 cccatctaca gctggcaggt ggccaagggc atcctgcccg accgcgccgt ggtgcagggc
     7441 aaggacgagg aggactggca gctgatcgac gacagcttca acttcaagtt cagcctgcac
     7501 cccaacgacc tggtggaggt gatcaccaag aaggcccgca tgttcggcta cttcgccagc
     7561 tgccaccgcg gcaccggcaa catcaacatc cgcatccacg acctggacca caagatcggc
     7621 aagaacggca tcctggaggg catcggcgtg aagaccgccc tgagcttcca gaagtaccag
     7681 atcgacgagc tgggcaagga gatccgcccc tgccgcctga agaagcgccc tcctgtgcgc
     7741 agcagggctg accccaagaa gaagaggaag gtggaggcca gcctcgagaa aaggccggcg
     7801 gccacgaaaa aggccggcca ggcaaaaaag aaaaagggca gttaatgatc tagtaatgat
     7861 ctagaactat agtgagtcgt attacgtaga tccagacatg ataagataca ttgatgagtt
     7921 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     7981 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     8041 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaattcg cggccgcggc
     8101 gccaatgcat tgggcccggt acccagcttt tgttcccttt agtgagggtt aattgcgcgc
     8161 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     8221 cacaacatac gagccgggag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     8281 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     8341 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     8401 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     8461 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     8521 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     8581 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     8641 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     8701 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     8761 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     8821 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     8881 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     8941 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     9001 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     9061 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     9121 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     9181 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     9241 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     9301 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     9361 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     9421 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     9481 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     9541 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     9601 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     9661 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     9721 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     9781 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     9841 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     9901 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     9961 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
    10021 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
    10081 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
    10141 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
    10201 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
    10261 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
    10321 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
    10381 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
    10441 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
    10501 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
    10561 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
    10621 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
    10681 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
    10741 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc
    10801 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagtcgaccg
    10861 ccaattcaat atggcgtata tggactcatg ccaattcaat atggtggatc tggacctgtg
    10921 ccaattcaat atggcgtata tggactcgtg ccaattcaat atggtggatc tggaccccag
    10981 ccaattcaat atggcggact tggcaccatg ccaattcaat atggcggacc tggcactgtg
    11041 ccaactgggg aggggtctac ttggcacggt gccaagtttg aggaggggtc ttggccctgt
    11101 gccaagtccg ccatattgaa ttggcatggt gccaataatg gcggccatat tggctatatg
    11161 ccaggatcaa tatataggca atatccaata tggccctatg ccaatatggc tattggccag
    11221 gttcaatact atgtattggc cctatgccat atagtattcc atatatgggt tttcctattg
    11281 acgtagatag cccctcccaa tgggcggtcc catataccat atatggggct tcctaatacc
    11341 gcccatagcc actcccccat tgacgtcaat ggtctctata tatggtcttt cctattgacg
    11401 tcatatgggc ggtcctattg acgtatatgg cgcctccccc attgacgtca attacggtaa
    11461 atggcccgcc tggctcaatg cccattgacg tcaataggac cacccaccat tgacgtcaat
    11521 gggatggctc attgcccatt catatccgtt ctcacgcccc ctattgacgt caatgacggt
    11581 aaatggccca cttggcagta catcaatatc tattaatagt aacttggcaa gtacattact
    11641 attggaagta cgccagggta cattggcagt actcccattg acgtcaatgg cggtaaatgg
    11701 cccgcgatgg ctgccaagta catccccatt gacgtcaatg gggaggggca atgacgcaaa
    11761 tgggcgttcc attgacgtaa atgggcggta ggcgtgccta atgggaggtc tatataagca
    11821 atgctcgttt aggg
//
