LOCUS       sequence-204682-       11751 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SpCas9 fused to dSaCas9 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11751)
  AUTHORS   Bolukbasi MF, Liu P, Luk K, Kwok SF, Gupta A, Amrani N, Sontheimer 
            EJ, Zhu LJ, Wolfe SA
  TITLE     Orthogonal Cas9-Cas9 chimeras provide a versatile platform for 
            genome editing.
  JOURNAL   Nat Commun. 2018 Nov 19;9(1):4856. doi: 10.1038/s41467-018-07310-x.
  PUBMED    30451839
REFERENCE   2  (bases 1 to 11751)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2018-11-19"; volume: "9(1):4856. doi"; pages: " 
            10.1038/s41467-018-07310-x"
COMMENT     Sequence Label: pCSD_2xNLS-SpCas9WT-NLS-3xHA-NLS-dSaCas9-2xNLS
FEATURES             Location/Qualifiers
     source          1..11751
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        221..239
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     221..238
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             289..336
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             355..375
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             382..4482
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             382..4482
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             382..4482
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4501..4521
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4522..4611
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     CDS             4624..4650
                     /codon_start=1
                     /product="nuclear localization signal (Makkerh et al., 
                     1996)"
                     /label=NLS
                     /translation="PAAKKKKLD"
     CDS             7855..7902
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7921..7947
                     /codon_start=1
                     /product="nuclear localization signal (Makkerh et al., 
                     1996)"
                     /label=NLS
                     /translation="PAAKKKKLD"
     polyA_signal    7989..8123
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8013..8032)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     8067..8086
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(8236..8256)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(8236..8254)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(8275..8291)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8275..8291)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8288..8310)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8299..8315
                     /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(8323..8329,8330..8347,8348..8353))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8368..8389
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8506..8523)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8677..9265)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8757..8776)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9436..10227,10228..10296))
                     /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     10059..10078
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10297..10401)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(10427..10882)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10564..10585)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     10776..10795
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(10953..11751,1..186)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 gccagggtac attggcagta ctcccattga cgtcaatggc ggtaaatggc ccgcgatggc
       61 tgccaagtac atccccattg acgtcaatgg ggaggggcaa tgacgcaaat gggcgttcca
      121 ttgacgtaaa tgggcggtag gcgtgcctaa tgggaggtct atataagcaa tgctcgttta
      181 gggaaccgcc attctgcctg gggacgtcgg agcaagcttg atttaggtga cactatagaa
      241 tacaagctac ttgttctttt tgcaggatcc catcgattcg aattcatgaa aaggccggcg
      301 gccacgaaaa aggccggcca ggcaaaaaag aaaaagctgg acggcagcgg aagccccaag
      361 aagaagagga aggtaccatg ggacaagaag tactccattg ggctcgatat cggtaccaac
      421 agcgtcggct gggccgtcat tacggacgag tacaaggtgc cgagcaaaaa attcaaagtt
      481 ctgggcaata ccgatcgcca cagcataaag aagaacctca ttggagccct cctgttcgac
      541 tccggggaga cggccgaagc cacgcggctc aaaagaacag cacggcgcag atatacccgc
      601 agaaagaatc ggatctgcta cctgcaggag atctttagta atgagatggc taaggtggat
      661 gactctttct tccataggct ggaggagtcc tttttggtgg aggaggataa aaagcacgag
      721 cgccacccaa tctttggcaa tatcgtggac gaggtggcgt accatgaaaa gtacccaacc
      781 atatatcatc tgaggaagaa gctggtagac agtactgata aggctgactt gcggttgatc
      841 tatctcgcgc tggcgcacat gatcaaattt cggggacact tcctcatcga gggggacctg
      901 aacccagaca acagcgatgt cgacaaactc tttatccaac tggttcagac ttacaatcag
      961 cttttcgagg agaacccgat caacgcatcc ggcgttgacg ccaaagcaat cctgagcgct
     1021 aggctgtcca aatcccggcg gctcgaaaac ctcatcgcac agctccctgg ggagaagaag
     1081 aacggcctgt ttggtaatct tatcgccctg tcactcgggc tgacccccaa ctttaaatct
     1141 aacttcgacc tggccgaaga tgccaagctg caactgagca aagacaccta cgatgatgat
     1201 ctcgacaatc tgctggccca gatcggcgac cagtacgcag accttttttt ggcggcaaag
     1261 aacctgtcag acgccattct gctgagtgat attctgcgag tgaacacgga gatcaccaaa
     1321 gctccgctga gcgctagtat gatcaagcgc tatgatgagc accaccaaga cttgactttg
     1381 ctgaaggccc ttgtcagaca gcaactgcct gagaagtaca aggaaatttt cttcgatcag
     1441 tctaaaaatg gctacgccgg atacattgac ggcggagcaa gccaggagga attttacaaa
     1501 tttattaagc ccatcttgga aaaaatggac ggcaccgagg agctgctggt aaagctgaac
     1561 agagaagatc tgttgcgcaa acagcgcact ttcgacaatg gaagcatccc ccaccagatt
     1621 cacctgggcg aactgcacgc tatcctcagg cggcaagagg atttctaccc ctttttgaaa
     1681 gataacaggg aaaagattga gaaaatcctc acatttcgga taccctacta tgtaggcccc
     1741 ctcgctcggg gaaattccag attcgcgtgg atgactcgca aatcagaaga gaccatcact
     1801 ccctggaact tcgaggaagt cgtggataag ggggcctctg cccagtcctt catcgaaagg
     1861 atgactaact ttgataaaaa tctgcctaac gaaaaggtgc ttcctaaaca ctctctgctg
     1921 tacgagtact tcacagttta taacgagctc accaaggtca aatacgtcac agaagggatg
     1981 agaaagccag cattcctgtc tggagagcag aagaaagcta tcgtggacct cctcttcaag
     2041 acgaaccgga aagttaccgt gaaacagctc aaagaagact atttcaaaaa gattgaatgt
     2101 ttcgactctg ttgaaatcag cggagtggag gatcgcttca acgcatccct gggaacgtat
     2161 cacgatctcc tgaaaatcat taaagacaag gacttcctgg acaatgagga gaacgaggac
     2221 attcttgagg acattgtcct cacccttacg ttgtttgaag atagggagat gattgaagaa
     2281 cgcttgaaaa cttacgctca tctcttcgac gacaaagtca tgaaacagct caagagacgc
     2341 cgatatacag gatgggggcg gctgtcaaga aaactgatca atggcatccg agacaagcag
     2401 agtggaaaga caatcctgga ttttcttaag tccgatggat ttgccaaccg gaacttcatg
     2461 cagttgatcc atgatgactc tctcaccttt aaggaggaca tccagaaagc acaagtttct
     2521 ggccaggggg acagtcttca cgagcacatc gctaatcttg caggtagccc agctatcaaa
     2581 aagggaatac tgcagaccgt taaggtcgtg gatgaactcg tcaaagtaat gggaaggcat
     2641 aagcccgaga atatcgttat cgagatggcc cgagagaacc aaactaccca gaagggacag
     2701 aagaacagta gggaaaggat gaagaggatt gaagagggta taaaagaact ggggtcccaa
     2761 atccttaagg aacacccagt tgaaaacacc cagcttcaga atgagaagct ctacctgtac
     2821 tacctgcaga acggcaggga catgtacgtg gatcaggaac tggacatcaa ccggttgtcc
     2881 gactacgacg tggatcatat cgtgccccaa agctttctca aagatgattc tattgataat
     2941 aaagtgttga caagatccga taaaaataga gggaagagtg ataacgtccc ctcagaagaa
     3001 gttgtcaaga aaatgaaaaa ttattggcgg cagctgctga acgccaaact gatcacacaa
     3061 cggaagttcg ataatctgac taaggctgaa cgaggtggcc tgtctgagtt ggataaagcc
     3121 ggcttcatca aaaggcagct tgttgagaca cgccagatca ccaagcacgt ggcccaaatt
     3181 ctcgattcac gcatgaacac caagtacgat gaaaatgaca aactgattcg agaggtgaaa
     3241 gttattactc tgaagtctaa gctggtctca gatttcagaa aggactttca gttttataag
     3301 gtgagagaga tcaacaatta ccaccatgcg catgatgcct acctgaatgc agtggtaggc
     3361 actgcactta tcaaaaaata tcccaagctg gaatctgaat ttgtttacgg agactataaa
     3421 gtgtacgatg ttaggaaaat gatcgcaaag tctgagcagg aaataggcaa ggccaccgct
     3481 aagtacttct tttacagcaa tattatgaat tttttcaaga ccgagattac actggccaat
     3541 ggagagattc ggaagcgacc acttatcgaa acaaacggag aaacaggaga aatcgtgtgg
     3601 gacaagggta gggatttcgc gacagtccgc aaggtcctgt ccatgccgca ggtgaacatc
     3661 gttaaaaaga ccgaagtaca gaccggaggc ttctccaagg aaagtatcct cccgaaaagg
     3721 aacagcgaca agctgatcgc acgcaaaaaa gattgggacc ccaagaaata cggcggattc
     3781 gattctccta cagtcgctta cagtgtactg gttgtggcca aagtggagaa agggaagtct
     3841 aaaaaactca aaagcgtcaa ggaactgctg ggcatcacaa tcatggagcg atccagcttc
     3901 gagaaaaacc ccatcgactt tctcgaagcg aaaggatata aagaggtcaa aaaagacctc
     3961 atcattaagc tgcccaagta ctctctcttt gagcttgaaa acggccggaa acgaatgctc
     4021 gctagtgcgg gcgagctgca gaaaggtaac gagctggcac tgccctctaa atacgttaat
     4081 ttcttgtatc tggccagcca ctatgaaaag ctcaaagggt ctcccgaaga taatgagcag
     4141 aagcagctgt tcgtggaaca acacaaacac taccttgatg agatcatcga gcaaataagc
     4201 gagttctcca aaagagtgat cctcgccgac gctaacctcg ataaggtgct ttctgcttac
     4261 aataagcaca gggataagcc catcagggag caggcagaaa acattatcca cttgtttact
     4321 ctgaccaact tgggcgcgcc tgcagccttc aagtacttcg acaccaccat agacagaaag
     4381 cggtacacct ctacaaagga ggtcctggac gccacactga ttcatcagtc aattacgggg
     4441 ctctatgaaa caagaatcga cctctctcag ctcggtggag acgctagcgg caccggcggg
     4501 cccaagaaga agaggaaggt atacccatac gatgttcctg actatgcggg ctatccctat
     4561 gacgtcccgg actatgcagg atcgtatcct tatgacgttc cagattacgc tggatccgcc
     4621 gctccggcag ctaagaaaaa gaaactggat ttcgaatccg gaactagtgg tatccacgga
     4681 gtcccagcag ccaagcggaa ctacatcctg ggcctggcca tcggcatcac cagcgtgggc
     4741 tacggcatca tcgactacga gacacgggac gtgatcgatg ccggcgtgcg gctgttcaaa
     4801 gaggccaacg tggaaaacaa cgagggcagg cggagcaaga gaggcgccag aaggctgaag
     4861 cggcggaggc ggcatagaat ccagagagtg aagaagctgc tgttcgacta caacctgctg
     4921 accgaccaca gcgagctgag cggcatcaac ccctacgagg ccagagtgaa gggcctgagc
     4981 cagaagctga gcgaggaaga gttctctgcc gccctgctgc acctggccaa gagaagaggc
     5041 gtgcacaacg tgaacgaggt ggaagaggac accggcaacg agctgtccac caaagagcag
     5101 atcagccgga acagcaaggc cctggaagag aaatacgtgg ccgaactgca gctggaacgg
     5161 ctgaagaaag acggcgaagt gcggggcagc atcaacagat tcaagaccag cgactacgtg
     5221 aaagaagcca aacagctgct gaaggtgcag aaggcctacc accagctgga ccagagcttc
     5281 atcgacacct acatcgacct gctggaaacc cggcggacct actatgaggg acctggcgag
     5341 ggcagcccct tcggctggaa ggacatcaaa gaatggtacg agatgctgat gggccactgc
     5401 acctacttcc ccgaggaact gcggagcgtg aagtacgcct acaacgccga cctgtacaac
     5461 gccctgaacg acctgaacaa tctcgtgatc accagggacg agaacgagaa gctggaatat
     5521 tacgagaagt tccagatcat cgagaacgtg ttcaagcaga agaagaagcc caccctgaag
     5581 cagatcgcca aagaaatcct cgtgaacgaa gaggatatta agggctacag agtgaccagc
     5641 accggcaagc ccgagttcac caacctgaag gtgtaccacg acatcaagga cattaccgcc
     5701 cggaaagaga ttattgagaa cgccgagctg ctggatcaga ttgccaagat cctgaccatc
     5761 taccagagca gcgaggacat ccaggaagaa ctgaccaatc tgaactccga gctgacccag
     5821 gaagagatcg agcagatctc taatctgaag ggctataccg gcacccacaa cctgagcctg
     5881 aaggccatca acctgatcct ggacgagctg tggcacacca acgacaacca gatcgctatc
     5941 ttcaaccggc tgaagctggt gcccaagaag gtggacctgt cccagcagaa agagatcccc
     6001 accaccctgg tggacgactt catcctgagc cccgtcgtga agagaagctt catccagagc
     6061 atcaaagtga tcaacgccat catcaagaag tacggcctgc ccaacgacat cattatcgag
     6121 ctggcccgcg agaagaactc caaggacgcc cagaaaatga tcaacgagat gcagaagcgg
     6181 aaccggcaga ccaacgagcg gatcgaggaa atcatccgga ccaccggcaa agagaacgcc
     6241 aagtacctga tcgagaagat caagctgcac gacatgcagg aaggcaagtg cctgtacagc
     6301 ctggaagcca tccctctgga agatctgctg aacaacccct tcaactatga ggtggaccac
     6361 atcatcccca gaagcgtgtc cttcgacaac agcttcaaca acaaggtgct cgtgaagcag
     6421 gaagaagcca gcaagaaggg caaccggacc ccattccagt acctgagcag cagcgacagc
     6481 aagatcagct acgaaacctt caagaagcac atcctgaatc tggccaaggg caagggcaga
     6541 atcagcaaga ccaagaaaga gtatctgctg gaagaacggg acatcaacag gttctccgtg
     6601 cagaaagact tcatcaaccg gaacctggtg gataccagat acgccaccag aggcctgatg
     6661 aacctgctgc ggagctactt cagagtgaac aacctggacg tgaaagtgaa gtccatcaat
     6721 ggcggcttca ccagctttct gcggcggaag tggaagttta agaaagagcg gaacaagggg
     6781 tacaagcacc acgccgagga cgccctgatc attgccaacg ccgatttcat cttcaaagag
     6841 tggaagaaac tggacaaggc caaaaaagtg atggaaaacc agatgttcga ggaaaagcag
     6901 gccgagagca tgcccgagat cgaaaccgag caggagtaca aagagatctt catcaccccc
     6961 caccagatca agcacattaa ggacttcaag gactacaagt acagccaccg ggtggacaag
     7021 aagcctaata gagagctgat taacgacacc ctgtactcca cccggaagga cgacaagggc
     7081 aacaccctga tcgtgaacaa tctgaacggc ctgtacgaca aggacaatga caagctgaaa
     7141 aagctgatca acaagagccc cgaaaagctg ctgatgtacc accacgaccc ccagacctac
     7201 cagaaactga agctgattat ggaacagtac ggcgacgaga agaatcccct gtacaagtac
     7261 tacgaggaaa ccgggaacta cctgaccaag tactccaaaa aggacaacgg ccccgtgatc
     7321 aagaagatta agtattacgg caacaaactg aacgcccatc tggacatcac cgacgactac
     7381 cccaacagca gaaacaaggt cgtgaagctg tccctgaagc cctacagatt cgacgtgtac
     7441 ctggacaatg gcgtgtacaa gttcgtgacc gtgaagaatc tggatgtgat caaaaaagaa
     7501 aactactacg aagtgaatag caagtgctat gaggaagcta agaagctgaa gaagatcagc
     7561 aaccaggccg agtttatcgc ctccttctac aacaacgatc tgatcaagat caacggcgag
     7621 ctgtatagag tgatcggcgt gaacaacgac ctgctgaacc ggatcgaagt gaacatgatc
     7681 gacatcacct accgcgagta cctggaaaac atgaacgaca agaggccccc caggatcatt
     7741 aagacaatcg cctccaagac ccagagcatt aagaagtaca gcacagacat tctgggcaac
     7801 ctgtatgaag tgaaatctaa gaagcaccct cagatcatca aaaagggccc cgggaaaagg
     7861 ccggcggcca cgaaaaaggc cggccaggca aaaaagaaaa agctggacgg cagcggaagc
     7921 ccggcagcta agaaaaagaa actggatctc gagtaataat ctagaactat agtgagtcgt
     7981 attacgtaga tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     8041 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     8101 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     8161 agggggaggt gtgggaggtt ttttaattcg cggccgcggc gccaatgcat tgggcccggt
     8221 acccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata
     8281 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccgggag
     8341 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     8401 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     8461 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     8521 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     8581 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     8641 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     8701 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     8761 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     8821 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     8881 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     8941 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     9001 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     9061 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     9121 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     9181 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     9241 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     9301 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     9361 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     9421 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     9481 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     9541 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     9601 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     9661 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     9721 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     9781 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     9841 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     9901 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     9961 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
    10021 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
    10081 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
    10141 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
    10201 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
    10261 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
    10321 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
    10381 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt
    10441 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
    10501 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
    10561 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
    10621 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
    10681 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
    10741 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
    10801 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
    10861 aatgcgccgc tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg
    10921 cgatcggtgc gggcctcttc gctattacgc cagtcgaccg ccaattcaat atggcgtata
    10981 tggactcatg ccaattcaat atggtggatc tggacctgtg ccaattcaat atggcgtata
    11041 tggactcgtg ccaattcaat atggtggatc tggaccccag ccaattcaat atggcggact
    11101 tggcaccatg ccaattcaat atggcggacc tggcactgtg ccaactgggg aggggtctac
    11161 ttggcacggt gccaagtttg aggaggggtc ttggccctgt gccaagtccg ccatattgaa
    11221 ttggcatggt gccaataatg gcggccatat tggctatatg ccaggatcaa tatataggca
    11281 atatccaata tggccctatg ccaatatggc tattggccag gttcaatact atgtattggc
    11341 cctatgccat atagtattcc atatatgggt tttcctattg acgtagatag cccctcccaa
    11401 tgggcggtcc catataccat atatggggct tcctaatacc gcccatagcc actcccccat
    11461 tgacgtcaat ggtctctata tatggtcttt cctattgacg tcatatgggc ggtcctattg
    11521 acgtatatgg cgcctccccc attgacgtca attacggtaa atggcccgcc tggctcaatg
    11581 cccattgacg tcaataggac cacccaccat tgacgtcaat gggatggctc attgcccatt
    11641 catatccgtt ctcacgcccc ctattgacgt caatgacggt aaatggccca cttggcagta
    11701 catcaatatc tattaatagt aacttggcaa gtacattact attggaagta c
//
