LOCUS       sequence-204749-       11834 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SpCas9 (R1335K) 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-SpCas9MT3-NLS-3xHA-2XNLS-NmCas9WT-2...
FEATURES             Location/Qualifiers
     source          1..11834
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        28..46
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     28..45
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             96..143
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             162..182
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             189..4289
                     /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
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKKYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4302..4322
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4323..4412
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     CDS             4425..4451
                     /codon_start=1
                     /product="nuclear localization signal (Makkerh et al., 
                     1996)"
                     /label=NLS
                     /translation="PAAKKKKLD"
     CDS             4467..4487
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4488..7730
                     /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      7317..7326
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7743..7763
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7779..7826
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     polyA_signal    7879..8013
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7903..7922)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7957..7976
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(8126..8146)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(8126..8144)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(8165..8181)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8165..8181)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8178..8200)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8189..8205
                     /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(8213..8219,8220..8237,8238..8243))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8258..8279
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8396..8413)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8567..9155)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8647..8666)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9326..10117,10118..10186))
                     /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     9949..9968
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10187..10291)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(10317..10772)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10454..10475)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     10666..10685
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        10843..11827
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 ctgcctgggg acgtcggagc aagcttgatt taggtgacac tatagaatac aagctacttg
       61 ttctttttgc aggatcccat cgattcgaat tcatgaaaag gccggcggcc acgaaaaagg
      121 ccggccaggc aaaaaagaaa aagctggacg gcagcggaag ccccaagaag aagaggaagg
      181 taccatggga caagaagtac tccattgggc tcgatatcgg taccaacagc gtcggctggg
      241 ccgtcattac ggacgagtac aaggtgccga gcaaaaaatt caaagttctg ggcaataccg
      301 atcgccacag cataaagaag aacctcattg gagccctcct gttcgactcc ggggagacgg
      361 ccgaagccac gcggctcaaa agaacagcac ggcgcagata tacccgcaga aagaatcgga
      421 tctgctacct gcaggagatc tttagtaatg agatggctaa ggtggatgac tctttcttcc
      481 ataggctgga ggagtccttt ttggtggagg aggataaaaa gcacgagcgc cacccaatct
      541 ttggcaatat cgtggacgag gtggcgtacc atgaaaagta cccaaccata tatcatctga
      601 ggaagaagct ggtagacagt actgataagg ctgacttgcg gttgatctat ctcgcgctgg
      661 cgcacatgat caaatttcgg ggacacttcc tcatcgaggg ggacctgaac ccagacaaca
      721 gcgatgtcga caaactcttt atccaactgg ttcagactta caatcagctt ttcgaggaga
      781 acccgatcaa cgcatccggc gttgacgcca aagcaatcct gagcgctagg ctgtccaaat
      841 cccggcggct cgaaaacctc atcgcacagc tccctgggga gaagaagaac ggcctgtttg
      901 gtaatcttat cgccctgtca ctcgggctga cccccaactt taaatctaac ttcgacctgg
      961 ccgaagatgc caagctgcaa ctgagcaaag acacctacga tgatgatctc gacaatctgc
     1021 tggcccagat cggcgaccag tacgcagacc tttttttggc ggcaaagaac ctgtcagacg
     1081 ccattctgct gagtgatatt ctgcgagtga acacggagat caccaaagct ccgctgagcg
     1141 ctagtatgat caagcgctat gatgagcacc accaagactt gactttgctg aaggcccttg
     1201 tcagacagca actgcctgag aagtacaagg aaattttctt cgatcagtct aaaaatggct
     1261 acgccggata cattgacggc ggagcaagcc aggaggaatt ttacaaattt attaagccca
     1321 tcttggaaaa aatggacggc accgaggagc tgctggtaaa gctgaacaga gaagatctgt
     1381 tgcgcaaaca gcgcactttc gacaatggaa gcatccccca ccagattcac ctgggcgaac
     1441 tgcacgctat cctcaggcgg caagaggatt tctacccctt tttgaaagat aacagggaaa
     1501 agattgagaa aatcctcaca tttcggatac cctactatgt aggccccctc gctcggggaa
     1561 attccagatt cgcgtggatg actcgcaaat cagaagagac catcactccc tggaacttcg
     1621 aggaagtcgt ggataagggg gcctctgccc agtccttcat cgaaaggatg actaactttg
     1681 ataaaaatct gcctaacgaa aaggtgcttc ctaaacactc tctgctgtac gagtacttca
     1741 cagtttataa cgagctcacc aaggtcaaat acgtcacaga agggatgaga aagccagcat
     1801 tcctgtctgg agagcagaag aaagctatcg tggacctcct cttcaagacg aaccggaaag
     1861 ttaccgtgaa acagctcaaa gaagactatt tcaaaaagat tgaatgtttc gactctgttg
     1921 aaatcagcgg agtggaggat cgcttcaacg catccctggg aacgtatcac gatctcctga
     1981 aaatcattaa agacaaggac ttcctggaca atgaggagaa cgaggacatt cttgaggaca
     2041 ttgtcctcac ccttacgttg tttgaagata gggagatgat tgaagaacgc ttgaaaactt
     2101 acgctcatct cttcgacgac aaagtcatga aacagctcaa gagacgccga tatacaggat
     2161 gggggcggct gtcaagaaaa ctgatcaatg gcatccgaga caagcagagt ggaaagacaa
     2221 tcctggattt tcttaagtcc gatggatttg ccaaccggaa cttcatgcag ttgatccatg
     2281 atgactctct cacctttaag gaggacatcc agaaagcaca agtttctggc cagggggaca
     2341 gtcttcacga gcacatcgct aatcttgcag gtagcccagc tatcaaaaag ggaatactgc
     2401 agaccgttaa ggtcgtggat gaactcgtca aagtaatggg aaggcataag cccgagaata
     2461 tcgttatcga gatggcccga gagaaccaaa ctacccagaa gggacagaag aacagtaggg
     2521 aaaggatgaa gaggattgaa gagggtataa aagaactggg gtcccaaatc cttaaggaac
     2581 acccagttga aaacacccag cttcagaatg agaagctcta cctgtactac ctgcagaacg
     2641 gcagggacat gtacgtggat caggaactgg acatcaaccg gttgtccgac tacgacgtgg
     2701 atcatatcgt gccccaaagc tttctcaaag atgattctat tgataataaa gtgttgacaa
     2761 gatccgataa aaatagaggg aagagtgata acgtcccctc agaagaagtt gtcaagaaaa
     2821 tgaaaaatta ttggcggcag ctgctgaacg ccaaactgat cacacaacgg aagttcgata
     2881 atctgactaa ggctgaacga ggtggcctgt ctgagttgga taaagccggc ttcatcaaaa
     2941 ggcagcttgt tgagacacgc cagatcacca agcacgtggc ccaaattctc gattcacgca
     3001 tgaacaccaa gtacgatgaa aatgacaaac tgattcgaga ggtgaaagtt attactctga
     3061 agtctaagct ggtctcagat ttcagaaagg actttcagtt ttataaggtg agagagatca
     3121 acaattacca ccatgcgcat gatgcctacc tgaatgcagt ggtaggcact gcacttatca
     3181 aaaaatatcc caagctggaa tctgaatttg tttacggaga ctataaagtg tacgatgtta
     3241 ggaaaatgat cgcaaagtct gagcaggaaa taggcaaggc caccgctaag tacttctttt
     3301 acagcaatat tatgaatttt ttcaagaccg agattacact ggccaatgga gagattcgga
     3361 agcgaccact tatcgaaaca aacggagaaa caggagaaat cgtgtgggac aagggtaggg
     3421 atttcgcgac agtccgcaag gtcctgtcca tgccgcaggt gaacatcgtt aaaaagaccg
     3481 aagtacagac cggaggcttc tccaaggaaa gtatcctccc gaaaaggaac agcgacaagc
     3541 tgatcgcacg caaaaaagat tgggacccca agaaatacgg cggattcgat tctcctacag
     3601 tcgcttacag tgtactggtt gtggccaaag tggagaaagg gaagtctaaa aaactcaaaa
     3661 gcgtcaagga actgctgggc atcacaatca tggagcgatc cagcttcgag aaaaacccca
     3721 tcgactttct cgaagcgaaa ggatataaag aggtcaaaaa agacctcatc attaagctgc
     3781 ccaagtactc tctctttgag cttgaaaacg gccggaaacg aatgctcgct agtgcgggcg
     3841 agctgcagaa aggtaacgag ctggcactgc cctctaaata cgttaatttc ttgtatctgg
     3901 ccagccacta tgaaaagctc aaagggtctc ccgaagataa tgagcagaag cagctgttcg
     3961 tggaacaaca caaacactac cttgatgaga tcatcgagca aataagcgag ttctccaaaa
     4021 gagtgatcct cgccgacgct aacctcgata aggtgctttc tgcttacaat aagcacaggg
     4081 ataagcccat cagggagcag gcagaaaaca ttatccactt gtttactctg accaacttgg
     4141 gcgcgcctgc agccttcaag tacttcgaca ccaccataga cagaaagaag tacacctcta
     4201 caaaggaggt cctggacgcc acactgattc atcagtcaat tacggggctc tatgaaacaa
     4261 gaatcgacct ctctcagctc ggtggagacg gcaccggcgg gcccaagaag aagaggaagg
     4321 tatacccata cgatgttcct gactatgcgg gctatcccta tgacgtcccg gactatgcag
     4381 gatcgtatcc ttatgacgtt ccagattacg ctggatccgc cgctccggca gctaagaaaa
     4441 agaaactgga tttcgaatcc ggaatgccca agaagaagag gaaggtggcc gccttcaagc
     4501 ccaaccccat caactacatc ctgggcctgg acatcggcat cgccagcgtg ggctgggcca
     4561 tggtggagat cgacgaggac gagaacccca tctgcctgat cgacctgggt gtgcgcgtgt
     4621 tcgagcgcgc tgaggtgccc aagactggtg acagtctggc tatggctcgc cggcttgctc
     4681 gctctgttcg gcgccttact cgccggcgcg ctcaccgcct tctgcgcgct cgccgcctgc
     4741 tgaagcgcga gggtgtgctg caggctgccg acttcgacga gaacggcctg atcaagagcc
     4801 tgcccaacac tccttggcag ctgcgcgctg ccgctctgga ccgcaagctg actcctctgg
     4861 agtggagcgc cgtgctgctg cacctgatca agcaccgcgg ctacctgagc cagcgcaaga
     4921 acgagggcga gaccgccgac aaggagctgg gtgctctgct gaagggcgtg gccgacaacg
     4981 cccacgccct gcagactggt gacttccgca ctcctgctga gctggccctg aacaagttcg
     5041 agaaggagag cggccacatc cgcaaccagc gcggcgacta cagccacacc ttcagccgca
     5101 aggacctgca ggccgagctg atcctgctgt tcgagaagca gaaggagttc ggcaaccccc
     5161 acgtgagcgg cggcctgaag gagggcatcg agaccctgct gatgacccag cgccccgccc
     5221 tgagcggcga cgccgtgcag aagatgctgg gccactgcac cttcgagcca gccgagccca
     5281 aggccgccaa gaacacctac accgccgagc gcttcatctg gctgaccaag ctgaacaacc
     5341 tgcgcatcct ggagcagggc agcgagcgcc ccctgaccga caccgagcgc gccaccctga
     5401 tggacgagcc ctaccgcaag agcaagctga cctacgccca ggcccgcaag ctgctgggtc
     5461 tggaggacac cgccttcttc aagggcctgc gctacggcaa ggacaacgcc gaggccagca
     5521 ccctgatgga gatgaaggcc taccacgcca tcagccgcgc cctggagaag gagggcctga
     5581 aggacaagaa gagtcctctg aacctgagcc ccgagctgca ggacgagatc ggcaccgcct
     5641 tcagcctgtt caagaccgac gaggacatca ccggccgcct gaaggaccgc atccagcccg
     5701 agatcctgga ggccctgctg aagcacatca gcttcgacaa gttcgtgcag atcagcctga
     5761 aggccctgcg ccgcatcgtg cccctgatgg agcagggcaa gcgctacgac gaggcctgcg
     5821 ccgagatcta cggcgaccac tacggcaaga agaacaccga ggagaagatc tacctgcctc
     5881 ctatccccgc cgacgagatc cgcaaccccg tggtgctgcg cgccctgagc caggcccgca
     5941 aggtgatcaa cggcgtggtg cgccgctacg gcagccccgc ccgcatccac atcgagaccg
     6001 cccgcgaggt gggcaagagc ttcaaggacc gcaaggagat cgagaagcgc caggaggaga
     6061 accgcaagga ccgcgagaag gccgccgcca agttccgcga gtacttcccc aacttcgtgg
     6121 gcgagcccaa gagcaaggac atcctgaagc tgcgcctgta cgagcagcag cacggcaagt
     6181 gcctgtacag cggcaaggag atcaacctgg gccgcctgaa cgagaagggc tacgtggaga
     6241 tcgaccatgc cctgcccttc agccgcacct gggacgacag cttcaacaac aaggtgctgg
     6301 tgctgggcag cgagaatcag aacaagggca accagacccc ctacgagtac ttcaacggca
     6361 aggacaacag ccgcgagtgg caggagttca aggcccgcgt ggagaccagc cgcttccccc
     6421 gcagcaagaa gcagcgcatc ctgctgcaga agttcgacga ggacggcttc aaggagcgca
     6481 acctgaacga cacccgctac gtgaaccgct tcctgtgcca gttcgtggcc gaccgcatgc
     6541 gcctgaccgg caagggcaag aagcgcgtgt tcgccagcaa cggccagatc accaacctgc
     6601 tgcgcggctt ctggggcctg cgcaaggtgc gcgccgagaa cgaccgccac cacgccctgg
     6661 acgccgtggt ggtggcctgc agcaccgtgg ccatgcagca gaagatcacc cgcttcgtgc
     6721 gctacaagga gatgaacgcc ttcgacggta aaaccatcga caaggagacc ggcgaggtgc
     6781 tgcaccagaa gacccacttc ccccagccct gggagttctt cgcccaggag gtgatgatcc
     6841 gcgtgttcgg caagcccgac ggcaagcccg agttcgagga ggccgacacc cccgagaagc
     6901 tgcgcaccct gctggccgag aagctgagca gccgccctga ggccgtgcac gagtacgtga
     6961 ctcctctgtt cgtgagccgc gcccccaacc gcaagatgag cggtcagggt cacatggaga
     7021 ccgtgaagag cgccaagcgc ctggacgagg gcgtgagcgt gctgcgcgtg cccctgaccc
     7081 agctgaagct gaaggacctg gagaagatgg tgaaccgcga gcgcgagccc aagctgtacg
     7141 aggccctgaa ggcccgcctg gaggcccaca aggacgaccc cgccaaggcc ttcgccgagc
     7201 ccttctacaa gtacgacaag gccggcaacc gcacccagca ggtgaaggcc gtgcgcgtgg
     7261 agcaggtgca gaagaccggc gtgtgggtgc gcaaccacaa cggcatcgcc gacaacgcca
     7321 ccatggtgcg cgtggacgtg ttcgagaagg gcgacaagta ctacctggtg cccatctaca
     7381 gctggcaggt ggccaagggc atcctgcccg accgcgccgt ggtgcagggc aaggacgagg
     7441 aggactggca gctgatcgac gacagcttca acttcaagtt cagcctgcac cccaacgacc
     7501 tggtggaggt gatcaccaag aaggcccgca tgttcggcta cttcgccagc tgccaccgcg
     7561 gcaccggcaa catcaacatc cgcatccacg acctggacca caagatcggc aagaacggca
     7621 tcctggaggg catcggcgtg aagaccgccc tgagcttcca gaagtaccag atcgacgagc
     7681 tgggcaagga gatccgcccc tgccgcctga agaagcgccc tcctgtgcgc agcagggctg
     7741 accccaagaa gaagaggaag gtggaggcca gcctcgagaa aaggccggcg gccacgaaaa
     7801 aggccggcca ggcaaaaaag aaaaagggca gttaatgatc tagtaatgat ctagaactat
     7861 agtgagtcgt attacgtaga tccagacatg ataagataca ttgatgagtt tggacaaacc
     7921 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     7981 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     8041 tttcaggttc agggggaggt gtgggaggtt ttttaattcg cggccgcggc gccaatgcat
     8101 tgggcccggt acccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat
     8161 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     8221 gagccgggag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     8281 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     8341 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     8401 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     8461 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     8521 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     8581 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     8641 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     8701 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     8761 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     8821 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     8881 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     8941 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     9001 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     9061 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     9121 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     9181 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     9241 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     9301 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     9361 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     9421 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     9481 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     9541 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     9601 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     9661 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     9721 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     9781 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     9841 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     9901 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     9961 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
    10021 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
    10081 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
    10141 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
    10201 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
    10261 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat
    10321 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
    10381 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
    10441 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
    10501 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
    10561 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
    10621 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
    10681 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
    10741 cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc tgcgcaactg
    10801 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagtcgaccg ccaattcaat
    10861 atggcgtata tggactcatg ccaattcaat atggtggatc tggacctgtg ccaattcaat
    10921 atggcgtata tggactcgtg ccaattcaat atggtggatc tggaccccag ccaattcaat
    10981 atggcggact tggcaccatg ccaattcaat atggcggacc tggcactgtg ccaactgggg
    11041 aggggtctac ttggcacggt gccaagtttg aggaggggtc ttggccctgt gccaagtccg
    11101 ccatattgaa ttggcatggt gccaataatg gcggccatat tggctatatg ccaggatcaa
    11161 tatataggca atatccaata tggccctatg ccaatatggc tattggccag gttcaatact
    11221 atgtattggc cctatgccat atagtattcc atatatgggt tttcctattg acgtagatag
    11281 cccctcccaa tgggcggtcc catataccat atatggggct tcctaatacc gcccatagcc
    11341 actcccccat tgacgtcaat ggtctctata tatggtcttt cctattgacg tcatatgggc
    11401 ggtcctattg acgtatatgg cgcctccccc attgacgtca attacggtaa atggcccgcc
    11461 tggctcaatg cccattgacg tcaataggac cacccaccat tgacgtcaat gggatggctc
    11521 attgcccatt catatccgtt ctcacgcccc ctattgacgt caatgacggt aaatggccca
    11581 cttggcagta catcaatatc tattaatagt aacttggcaa gtacattact attggaagta
    11641 cgccagggta cattggcagt actcccattg acgtcaatgg cggtaaatgg cccgcgatgg
    11701 ctgccaagta catccccatt gacgtcaatg gggaggggca atgacgcaaa tgggcgttcc
    11761 attgacgtaa atgggcggta ggcgtgccta atgggaggtc tatataagca atgctcgttt
    11821 agggaaccgc catt
//
