LOCUS       sequence-204717-       11834 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SpCas9 (R1335K) fused to dNmCas9 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-NmdCas9-2XNLS
FEATURES             Location/Qualifiers
     source          1..11834
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             48..95
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             114..134
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             141..4241
                     /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             4254..4274
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4275..4364
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     CDS             4377..4403
                     /codon_start=1
                     /product="nuclear localization signal (Makkerh et al., 
                     1996)"
                     /label=NLS
                     /translation="PAAKKKKLD"
     CDS             4419..4439
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(4440..4481,4482..4484,4485..6194,6195..6200,6201..6266,
                     6267..6269,6270..7682)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Neisseria meningitidis Type II 
                     CRISPR/Cas system"
                     /label=NmdCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the mutations D16A, D587A, 
                     H588A, and N611A"
                     /translation="AAFKPNPINYILGLAIGIASVGWAMVEIDEDENPICLIDLGVRVF
                     ERAEVPKTGDSLAMARRLARSVRRLTRRRAHRLLRARRLLKREGVLQAADFDENGLIKS
                     LPNTPWQLRAAALDRKLTPLEWSAVLLHLIKHRGYLSQRKNEGETADKELGALLKGVAD
                     NAHALQTGDFRTPAELALNKFEKESGHIRNQRGDYSHTFSRKDLQAELILLFEKQKEFG
                     NPHVSGGLKEGIETLLMTQRPALSGDAVQKMLGHCTFEPAEPKAAKNTYTAERFIWLTK
                     LNNLRILEQGSERPLTDTERATLMDEPYRKSKLTYAQARKLLGLEDTAFFKGLRYGKDN
                     AEASTLMEMKAYHAISRALEKEGLKDKKSPLNLSPELQDEIGTAFSLFKTDEDITGRLK
                     DRIQPEILEALLKHISFDKFVQISLKALRRIVPLMEQGKRYDEACAEIYGDHYGKKNTE
                     EKIYLPPIPADEIRNPVVLRALSQARKVINGVVRRYGSPARIHIETAREVGKSFKDRKE
                     IEKRQEENRKDREKAAAKFREYFPNFVGEPKSKDILKLRLYEQQHGKCLYSGKEINLGR
                     LNEKGYVEIAAALPFSRTWDDSFNNKVLVLGSEAQNKGNQTPYEYFNGKDNSREWQEFK
                     ARVETSRFPRSKKQRILLQKFDEDGFKERNLNDTRYVNRFLCQFVADRMRLTGKGKKRV
                     FASNGQITNLLRGFWGLRKVRAENDRHHALDAVVVACSTVAMQQKITRFVRYKEMNAFD
                     GKTIDKETGEVLHQKTHFPQPWEFFAQEVMIRVFGKPDGKPEFEEADTPEKLRTLLAEK
                     LSSRPEAVHEYVTPLFVSRAPNRKMSGQGHMETVKSAKRLDEGVSVLRVPLTQLKLKDL
                     EKMVNREREPKLYEALKARLEAHKDDPAKAFAEPFYKYDKAGNRTQQVKAVRVEQVQKT
                     GVWVRNHNGIADNATMVRVDVFEKGDKYYLVPIYSWQVAKGILPDRAVVQGKDEEDWQL
                     IDDSFNFKFSLHPNDLVEVITKKARMFGYFASCHRGTGNINIRIHDLDHKIGKNGILEG
                     IGVKTALSFQKYQIDELGKEIRPCRLKKRPPVR"
     regulatory      7269..7278
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7695..7715
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7731..7778
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     polyA_signal    7831..7965
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7855..7874)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7909..7928
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(8078..8098)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(8078..8096)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(8117..8133)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8117..8133)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8130..8152)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8141..8157
                     /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(8165..8171,8172..8189,8190..8195))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8210..8231
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8348..8365)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8519..9107)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8599..8618)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9278..10069,10070..10138))
                     /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     9901..9920
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10139..10243)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(10269..10724)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10406..10427)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     10618..10637
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        10795..11779
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        11814..11832
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     11814..11831
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acaagctact tgttcttttt gcaggatccc atcgattcga attcatgaaa aggccggcgg
       61 ccacgaaaaa ggccggccag gcaaaaaaga aaaagctgga cggcagcgga agccccaaga
      121 agaagaggaa ggtaccatgg gacaagaagt actccattgg gctcgatatc ggtaccaaca
      181 gcgtcggctg ggccgtcatt acggacgagt acaaggtgcc gagcaaaaaa ttcaaagttc
      241 tgggcaatac cgatcgccac agcataaaga agaacctcat tggagccctc ctgttcgact
      301 ccggggagac ggccgaagcc acgcggctca aaagaacagc acggcgcaga tatacccgca
      361 gaaagaatcg gatctgctac ctgcaggaga tctttagtaa tgagatggct aaggtggatg
      421 actctttctt ccataggctg gaggagtcct ttttggtgga ggaggataaa aagcacgagc
      481 gccacccaat ctttggcaat atcgtggacg aggtggcgta ccatgaaaag tacccaacca
      541 tatatcatct gaggaagaag ctggtagaca gtactgataa ggctgacttg cggttgatct
      601 atctcgcgct ggcgcacatg atcaaatttc ggggacactt cctcatcgag ggggacctga
      661 acccagacaa cagcgatgtc gacaaactct ttatccaact ggttcagact tacaatcagc
      721 ttttcgagga gaacccgatc aacgcatccg gcgttgacgc caaagcaatc ctgagcgcta
      781 ggctgtccaa atcccggcgg ctcgaaaacc tcatcgcaca gctccctggg gagaagaaga
      841 acggcctgtt tggtaatctt atcgccctgt cactcgggct gacccccaac tttaaatcta
      901 acttcgacct ggccgaagat gccaagctgc aactgagcaa agacacctac gatgatgatc
      961 tcgacaatct gctggcccag atcggcgacc agtacgcaga cctttttttg gcggcaaaga
     1021 acctgtcaga cgccattctg ctgagtgata ttctgcgagt gaacacggag atcaccaaag
     1081 ctccgctgag cgctagtatg atcaagcgct atgatgagca ccaccaagac ttgactttgc
     1141 tgaaggccct tgtcagacag caactgcctg agaagtacaa ggaaattttc ttcgatcagt
     1201 ctaaaaatgg ctacgccgga tacattgacg gcggagcaag ccaggaggaa ttttacaaat
     1261 ttattaagcc catcttggaa aaaatggacg gcaccgagga gctgctggta aagctgaaca
     1321 gagaagatct gttgcgcaaa cagcgcactt tcgacaatgg aagcatcccc caccagattc
     1381 acctgggcga actgcacgct atcctcaggc ggcaagagga tttctacccc tttttgaaag
     1441 ataacaggga aaagattgag aaaatcctca catttcggat accctactat gtaggccccc
     1501 tcgctcgggg aaattccaga ttcgcgtgga tgactcgcaa atcagaagag accatcactc
     1561 cctggaactt cgaggaagtc gtggataagg gggcctctgc ccagtccttc atcgaaagga
     1621 tgactaactt tgataaaaat ctgcctaacg aaaaggtgct tcctaaacac tctctgctgt
     1681 acgagtactt cacagtttat aacgagctca ccaaggtcaa atacgtcaca gaagggatga
     1741 gaaagccagc attcctgtct ggagagcaga agaaagctat cgtggacctc ctcttcaaga
     1801 cgaaccggaa agttaccgtg aaacagctca aagaagacta tttcaaaaag attgaatgtt
     1861 tcgactctgt tgaaatcagc ggagtggagg atcgcttcaa cgcatccctg ggaacgtatc
     1921 acgatctcct gaaaatcatt aaagacaagg acttcctgga caatgaggag aacgaggaca
     1981 ttcttgagga cattgtcctc acccttacgt tgtttgaaga tagggagatg attgaagaac
     2041 gcttgaaaac ttacgctcat ctcttcgacg acaaagtcat gaaacagctc aagagacgcc
     2101 gatatacagg atgggggcgg ctgtcaagaa aactgatcaa tggcatccga gacaagcaga
     2161 gtggaaagac aatcctggat tttcttaagt ccgatggatt tgccaaccgg aacttcatgc
     2221 agttgatcca tgatgactct ctcaccttta aggaggacat ccagaaagca caagtttctg
     2281 gccaggggga cagtcttcac gagcacatcg ctaatcttgc aggtagccca gctatcaaaa
     2341 agggaatact gcagaccgtt aaggtcgtgg atgaactcgt caaagtaatg ggaaggcata
     2401 agcccgagaa tatcgttatc gagatggccc gagagaacca aactacccag aagggacaga
     2461 agaacagtag ggaaaggatg aagaggattg aagagggtat aaaagaactg gggtcccaaa
     2521 tccttaagga acacccagtt gaaaacaccc agcttcagaa tgagaagctc tacctgtact
     2581 acctgcagaa cggcagggac atgtacgtgg atcaggaact ggacatcaac cggttgtccg
     2641 actacgacgt ggatcatatc gtgccccaaa gctttctcaa agatgattct attgataata
     2701 aagtgttgac aagatccgat aaaaatagag ggaagagtga taacgtcccc tcagaagaag
     2761 ttgtcaagaa aatgaaaaat tattggcggc agctgctgaa cgccaaactg atcacacaac
     2821 ggaagttcga taatctgact aaggctgaac gaggtggcct gtctgagttg gataaagccg
     2881 gcttcatcaa aaggcagctt gttgagacac gccagatcac caagcacgtg gcccaaattc
     2941 tcgattcacg catgaacacc aagtacgatg aaaatgacaa actgattcga gaggtgaaag
     3001 ttattactct gaagtctaag ctggtctcag atttcagaaa ggactttcag ttttataagg
     3061 tgagagagat caacaattac caccatgcgc atgatgccta cctgaatgca gtggtaggca
     3121 ctgcacttat caaaaaatat cccaagctgg aatctgaatt tgtttacgga gactataaag
     3181 tgtacgatgt taggaaaatg atcgcaaagt ctgagcagga aataggcaag gccaccgcta
     3241 agtacttctt ttacagcaat attatgaatt ttttcaagac cgagattaca ctggccaatg
     3301 gagagattcg gaagcgacca cttatcgaaa caaacggaga aacaggagaa atcgtgtggg
     3361 acaagggtag ggatttcgcg acagtccgca aggtcctgtc catgccgcag gtgaacatcg
     3421 ttaaaaagac cgaagtacag accggaggct tctccaagga aagtatcctc ccgaaaagga
     3481 acagcgacaa gctgatcgca cgcaaaaaag attgggaccc caagaaatac ggcggattcg
     3541 attctcctac agtcgcttac agtgtactgg ttgtggccaa agtggagaaa gggaagtcta
     3601 aaaaactcaa aagcgtcaag gaactgctgg gcatcacaat catggagcga tccagcttcg
     3661 agaaaaaccc catcgacttt ctcgaagcga aaggatataa agaggtcaaa aaagacctca
     3721 tcattaagct gcccaagtac tctctctttg agcttgaaaa cggccggaaa cgaatgctcg
     3781 ctagtgcggg cgagctgcag aaaggtaacg agctggcact gccctctaaa tacgttaatt
     3841 tcttgtatct ggccagccac tatgaaaagc tcaaagggtc tcccgaagat aatgagcaga
     3901 agcagctgtt cgtggaacaa cacaaacact accttgatga gatcatcgag caaataagcg
     3961 agttctccaa aagagtgatc ctcgccgacg ctaacctcga taaggtgctt tctgcttaca
     4021 ataagcacag ggataagccc atcagggagc aggcagaaaa cattatccac ttgtttactc
     4081 tgaccaactt gggcgcgcct gcagccttca agtacttcga caccaccata gacagaaaga
     4141 agtacacctc tacaaaggag gtcctggacg ccacactgat tcatcagtca attacggggc
     4201 tctatgaaac aagaatcgac ctctctcagc tcggtggaga cggcaccggc gggcccaaga
     4261 agaagaggaa ggtataccca tacgatgttc ctgactatgc gggctatccc tatgacgtcc
     4321 cggactatgc aggatcgtat ccttatgacg ttccagatta cgctggatcc gccgctccgg
     4381 cagctaagaa aaagaaactg gatttcgaat ccggaatgcc caagaagaag aggaaggtgg
     4441 ccgccttcaa gcccaacccc atcaactaca tcctgggcct ggccatcggc atcgccagcg
     4501 tgggctgggc catggtggag atcgacgagg acgagaaccc catctgcctg atcgacctgg
     4561 gtgtgcgcgt gttcgagcgc gctgaggtgc ccaagactgg tgacagtctg gctatggctc
     4621 gccggcttgc tcgctctgtt cggcgcctta ctcgccggcg cgctcaccgc cttctgcgcg
     4681 ctcgccgcct gctgaagcgc gagggtgtgc tgcaggctgc cgacttcgac gagaacggcc
     4741 tgatcaagag cctgcccaac actccttggc agctgcgcgc tgccgctctg gaccgcaagc
     4801 tgactcctct ggagtggagc gccgtgctgc tgcacctgat caagcaccgc ggctacctga
     4861 gccagcgcaa gaacgagggc gagaccgccg acaaggagct gggtgctctg ctgaagggcg
     4921 tggccgacaa cgcccacgcc ctgcagactg gtgacttccg cactcctgct gagctggccc
     4981 tgaacaagtt cgagaaggag agcggccaca tccgcaacca gcgcggcgac tacagccaca
     5041 ccttcagccg caaggacctg caggccgagc tgatcctgct gttcgagaag cagaaggagt
     5101 tcggcaaccc ccacgtgagc ggcggcctga aggagggcat cgagaccctg ctgatgaccc
     5161 agcgccccgc cctgagcggc gacgccgtgc agaagatgct gggccactgc accttcgagc
     5221 cagccgagcc caaggccgcc aagaacacct acaccgccga gcgcttcatc tggctgacca
     5281 agctgaacaa cctgcgcatc ctggagcagg gcagcgagcg ccccctgacc gacaccgagc
     5341 gcgccaccct gatggacgag ccctaccgca agagcaagct gacctacgcc caggcccgca
     5401 agctgctggg tctggaggac accgccttct tcaagggcct gcgctacggc aaggacaacg
     5461 ccgaggccag caccctgatg gagatgaagg cctaccacgc catcagccgc gccctggaga
     5521 aggagggcct gaaggacaag aagagtcctc tgaacctgag ccccgagctg caggacgaga
     5581 tcggcaccgc cttcagcctg ttcaagaccg acgaggacat caccggccgc ctgaaggacc
     5641 gcatccagcc cgagatcctg gaggccctgc tgaagcacat cagcttcgac aagttcgtgc
     5701 agatcagcct gaaggccctg cgccgcatcg tgcccctgat ggagcagggc aagcgctacg
     5761 acgaggcctg cgccgagatc tacggcgacc actacggcaa gaagaacacc gaggagaaga
     5821 tctacctgcc tcctatcccc gccgacgaga tccgcaaccc cgtggtgctg cgcgccctga
     5881 gccaggcccg caaggtgatc aacggcgtgg tgcgccgcta cggcagcccc gcccgcatcc
     5941 acatcgagac cgcccgcgag gtgggcaaga gcttcaagga ccgcaaggag atcgagaagc
     6001 gccaggagga gaaccgcaag gaccgcgaga aggccgccgc caagttccgc gagtacttcc
     6061 ccaacttcgt gggcgagccc aagagcaagg acatcctgaa gctgcgcctg tacgagcagc
     6121 agcacggcaa gtgcctgtac agcggcaagg agatcaacct gggccgcctg aacgagaagg
     6181 gctacgtgga gatcgccgct gccctgccct tcagccgcac ctgggacgac agcttcaaca
     6241 acaaggtgct ggtgctgggc agcgaggctc agaacaaggg caaccagacc ccctacgagt
     6301 acttcaacgg caaggacaac agccgcgagt ggcaggagtt caaggcccgc gtggagacca
     6361 gccgcttccc ccgcagcaag aagcagcgca tcctgctgca gaagttcgac gaggacggct
     6421 tcaaggagcg caacctgaac gacacccgct acgtgaaccg cttcctgtgc cagttcgtgg
     6481 ccgaccgcat gcgcctgacc ggcaagggca agaagcgcgt gttcgccagc aacggccaga
     6541 tcaccaacct gctgcgcggc ttctggggcc tgcgcaaggt gcgcgccgag aacgaccgcc
     6601 accacgccct ggacgccgtg gtggtggcct gcagcaccgt ggccatgcag cagaagatca
     6661 cccgcttcgt gcgctacaag gagatgaacg ccttcgacgg taaaaccatc gacaaggaga
     6721 ccggcgaggt gctgcaccag aagacccact tcccccagcc ctgggagttc ttcgcccagg
     6781 aggtgatgat ccgcgtgttc ggcaagcccg acggcaagcc cgagttcgag gaggccgaca
     6841 cccccgagaa gctgcgcacc ctgctggccg agaagctgag cagccgccct gaggccgtgc
     6901 acgagtacgt gactcctctg ttcgtgagcc gcgcccccaa ccgcaagatg agcggtcagg
     6961 gtcacatgga gaccgtgaag agcgccaagc gcctggacga gggcgtgagc gtgctgcgcg
     7021 tgcccctgac ccagctgaag ctgaaggacc tggagaagat ggtgaaccgc gagcgcgagc
     7081 ccaagctgta cgaggccctg aaggcccgcc tggaggccca caaggacgac cccgccaagg
     7141 ccttcgccga gcccttctac aagtacgaca aggccggcaa ccgcacccag caggtgaagg
     7201 ccgtgcgcgt ggagcaggtg cagaagaccg gcgtgtgggt gcgcaaccac aacggcatcg
     7261 ccgacaacgc caccatggtg cgcgtggacg tgttcgagaa gggcgacaag tactacctgg
     7321 tgcccatcta cagctggcag gtggccaagg gcatcctgcc cgaccgcgcc gtggtgcagg
     7381 gcaaggacga ggaggactgg cagctgatcg acgacagctt caacttcaag ttcagcctgc
     7441 accccaacga cctggtggag gtgatcacca agaaggcccg catgttcggc tacttcgcca
     7501 gctgccaccg cggcaccggc aacatcaaca tccgcatcca cgacctggac cacaagatcg
     7561 gcaagaacgg catcctggag ggcatcggcg tgaagaccgc cctgagcttc cagaagtacc
     7621 agatcgacga gctgggcaag gagatccgcc cctgccgcct gaagaagcgc cctcctgtgc
     7681 gcagcagggc tgaccccaag aagaagagga aggtggaggc cagcctcgag aaaaggccgg
     7741 cggccacgaa aaaggccggc caggcaaaaa agaaaaaggg cagttaatga tctagtaatg
     7801 atctagaact atagtgagtc gtattacgta gatccagaca tgataagata cattgatgag
     7861 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     7921 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     7981 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaatt cgcggccgcg
     8041 gcgccaatgc attgggcccg gtacccagct tttgttccct ttagtgaggg ttaattgcgc
     8101 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     8161 cacacaacat acgagccggg agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     8221 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     8281 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     8341 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     8401 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     8461 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     8521 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     8581 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     8641 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     8701 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     8761 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     8821 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     8881 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     8941 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     9001 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     9061 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     9121 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     9181 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     9241 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     9301 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     9361 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     9421 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     9481 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     9541 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     9601 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     9661 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     9721 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     9781 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     9841 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     9901 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     9961 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
    10021 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
    10081 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
    10141 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
    10201 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
    10261 tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
    10321 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
    10381 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
    10441 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
    10501 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
    10561 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
    10621 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
    10681 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccattcag
    10741 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagtcgac
    10801 cgccaattca atatggcgta tatggactca tgccaattca atatggtgga tctggacctg
    10861 tgccaattca atatggcgta tatggactcg tgccaattca atatggtgga tctggacccc
    10921 agccaattca atatggcgga cttggcacca tgccaattca atatggcgga cctggcactg
    10981 tgccaactgg ggaggggtct acttggcacg gtgccaagtt tgaggagggg tcttggccct
    11041 gtgccaagtc cgccatattg aattggcatg gtgccaataa tggcggccat attggctata
    11101 tgccaggatc aatatatagg caatatccaa tatggcccta tgccaatatg gctattggcc
    11161 aggttcaata ctatgtattg gccctatgcc atatagtatt ccatatatgg gttttcctat
    11221 tgacgtagat agcccctccc aatgggcggt cccatatacc atatatgggg cttcctaata
    11281 ccgcccatag ccactccccc attgacgtca atggtctcta tatatggtct ttcctattga
    11341 cgtcatatgg gcggtcctat tgacgtatat ggcgcctccc ccattgacgt caattacggt
    11401 aaatggcccg cctggctcaa tgcccattga cgtcaatagg accacccacc attgacgtca
    11461 atgggatggc tcattgccca ttcatatccg ttctcacgcc ccctattgac gtcaatgacg
    11521 gtaaatggcc cacttggcag tacatcaata tctattaata gtaacttggc aagtacatta
    11581 ctattggaag tacgccaggg tacattggca gtactcccat tgacgtcaat ggcggtaaat
    11641 ggcccgcgat ggctgccaag tacatcccca ttgacgtcaa tggggagggg caatgacgca
    11701 aatgggcgtt ccattgacgt aaatgggcgg taggcgtgcc taatgggagg tctatataag
    11761 caatgctcgt ttagggaacc gccattctgc ctggggacgt cggagcaagc ttgatttagg
    11821 tgacactata gaat
//
