LOCUS       sequence-197502-        8070 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression plasmid for human codon-optimized dead/inactive SpCas9 
            (without U6-sgRNA coding sequence).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8070)
  AUTHORS   Kulcsar PI, Talas A, Huszar K, Ligeti Z, Toth E, Weinhardt N, Fodor 
            E, Welker E
  TITLE     Crossing enhanced and high fidelity SpCas9 nucleases to optimize 
            specificity and cleavage.
  JOURNAL   Genome Biol. 2017 Oct 6;18(1):190. doi: 10.1186/s13059-017-1318-8.
  PUBMED    28985763
REFERENCE   2  (bases 1 to 8070)
  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: "Genome 
            Biol."; date: "2017-10-6"; volume: "18(1):190. doi"; pages: " 
            10.1186/s13059-017-1318-8"
COMMENT     Sequence Label: pX330-Flag-dSpCas9
FEATURES             Location/Qualifiers
     source          1..8070
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..286
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        288..562
                     /label=chicken beta-actin promoter
     intron          563..790
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      802..811
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             811..876
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             883..903
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(928..951,952..954,955..3510,3511..3513,3514..5028)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the N863A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             join(928..951,952..954,955..3441,3442..3444,3445..5028)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5029..5076
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(5101..5118)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5107..5314
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5323..5463
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5323..5452
                     /label=AAV2 ITR
     rep_origin      5538..5993
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5625..5644)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5835..5856
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6010..6029)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6129..6151
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6189..6207)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6275..6379
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6380..6448,6449..7240)
                     /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     complement(6598..6617)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7411..7999
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7900..7919
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      121 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      181 cgtcaatgac ggtaaatggc ccgcctggca ttgtgcccag tacatgacct tatgggactt
      241 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtcg aggtgagccc
      301 cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt
      361 tattttttaa ttattttgtg cagcgatggg ggcggggggg ggggggcgcg cgccaggcgg
      421 ggcggggcgg ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg cagccaatca
      481 gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc ggccctataa
      541 aaagcgaagc gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc
      601 cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc gcgttactcc cacaggtgag
      661 cgggcgggac ggcccttctc ctccgggctg taattagctg agcaagaggt aagggtttaa
      721 gggatggttg gttggtgggg tattaatgtt taattacctg gagcacctgc ctgaaatcac
      781 tttttttcag gttggaccgg tgccaccatg gactataagg accacgacgg agactacaag
      841 gatcatgata ttgattacaa agacgatgac gataagatgg ccccaaagaa gaagcggaag
      901 gtcggtatcc acggagtccc agcagccgac aagaagtaca gcatcgggct agccatcggc
      961 accaactctg tgggctgggc cgtgatcacc gacgagtaca aggtgcccag caagaaattc
     1021 aaggtgctgg gcaacaccga ccggcacagc atcaagaaga acctgatcgg agccctgctg
     1081 ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga gaaccgccag aagaagatac
     1141 accagacgga agaaccggat ctgctatctg caagagatct tcagcaacga gatggccaag
     1201 gtggacgaca gcttcttcca cagactggaa gagtccttcc tggtggaaga ggataagaag
     1261 cacgagcggc accccatctt cggcaacatc gtggacgagg tggcctacca cgagaagtac
     1321 cccaccatct accacctgag aaagaaactg gtggacagca ccgacaaggc cgacctgcgg
     1381 ctgatctatc tggccctggc ccacatgatc aagttccggg gccacttcct gatcgagggc
     1441 gacctgaacc ccgacaacag cgacgtggac aagctgttca tccagctggt gcagacctac
     1501 aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa ggccatcctg
     1561 tctgccagac tgagcaagag cagacggctg gaaaatctga tcgcccagct gcccggcgag
     1621 aagaagaatg gcctgttcgg caacctgatt gccctgagcc tgggcctgac ccccaacttc
     1681 aagagcaact tcgacctggc cgaggatgcc aaactgcagc tgagcaagga cacctacgac
     1741 gacgacctgg acaacctgct ggcccagatc ggcgaccagt acgccgacct gtttctggcc
     1801 gccaagaacc tgtccgacgc catcctgctg agcgacatcc tgagagtgaa caccgagatc
     1861 accaaggccc ccctgagcgc ctctatgatc aagagatacg acgagcacca ccaggacctg
     1921 accctgctga aagctctcgt gcggcagcag ctgcctgaga agtacaaaga gattttcttc
     1981 gaccagagca agaacggcta cgccggctac attgacggcg gagccagcca ggaagagttc
     2041 tacaagttca tcaagcccat cctggaaaag atggacggca ccgaggaact gctcgtgaag
     2101 ctgaacagag aggacctgct gcggaagcag cggaccttcg acaacggcag catcccccac
     2161 cagatccacc tgggagagct gcacgccatt ctgcggcggc aggaagattt ttacccattc
     2221 ctgaaggaca accgggaaaa gatcgagaag atcctgacct tccgcatccc ctactacgtg
     2281 ggccctctgg ccaggggaaa cagcagattc gcctggatga ccagaaagag cgaggaaacc
     2341 atcaccccct ggaacttcga ggaagtggtg gacaagggcg cttccgccca gagcttcatc
     2401 gagcggatga ccaacttcga taagaacctg cccaacgaga aggtgctgcc caagcacagc
     2461 ctgctgtacg agtacttcac cgtgtataac gagctgacca aagtgaaata cgtgaccgag
     2521 ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa aggccatcgt ggacctgctg
     2581 ttcaagacca accggaaagt gaccgtgaag cagctgaaag aggactactt caagaaaatc
     2641 gagtgcttcg actccgtgga aatctccggc gtggaagatc ggttcaacgc ctccctgggc
     2701 acataccacg atctgctgaa aattatcaag gacaaggact tcctggacaa tgaggaaaac
     2761 gaggacattc tggaagatat cgtgctgacc ctgacactgt ttgaggacag agagatgatc
     2821 gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag
     2881 cggcggagat acaccggctg gggcaggctg agccggaagc tgatcaacgg catccgggac
     2941 aagcagtccg gcaagacaat cctggatttc ctgaagtccg acggcttcgc caacagaaac
     3001 ttcatgcagc tgatccacga cgacagcctg acctttaaag aggacatcca gaaagcccag
     3061 gtgtccggcc agggcgatag cctgcacgag cacattgcca atctggccgg cagccccgcc
     3121 attaagaagg gcatcctgca gacagtgaag gtggtggacg agctcgtgaa agtgatgggc
     3181 cggcacaagc ccgagaacat cgtgatcgaa atggccagag agaaccagac cacccagaag
     3241 ggacagaaga acagccgcga gagaatgaag cggatcgaag agggcatcaa agagctgggc
     3301 agccagatcc tgaaagaaca ccccgtggaa aacacccagc tgcagaacga gaagctgtac
     3361 ctgtactacc tgcagaatgg gcgggatatg tacgtggacc aggaactgga catcaaccgg
     3421 ctgtccgact acgatgtgga cgccatcgtg cctcagagct ttctgaagga cgactccatc
     3481 gacaacaagg tgctgaccag aagcgacaag aaccggggca agagcgacaa cgtgccctcc
     3541 gaagaggtcg tgaagaagat gaagaactac tggcggcagc tgctgaacgc caagctgatt
     3601 acccagagaa agttcgacaa tctgaccaag gccgagagag gcggcctgag cgaactggat
     3661 aaggccggct tcatcaagag acagctggtg gaaacccggc agatcacaaa gcacgtggca
     3721 cagatcctgg actcccggat gaacactaag tacgacgaga atgacaagct gatccgggaa
     3781 gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt tccggaagga tttccagttt
     3841 tacaaagtgc gcgagatcaa caactaccac cacgcccacg acgcctacct gaacgccgtc
     3901 gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt gtacggcgac
     3961 tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg agcaggaaat cggcaaggct
     4021 accgccaagt acttcttcta cagcaacatc atgaactttt tcaagaccga gattaccctg
     4081 gccaacggcg agatccggaa gcggcctctg atcgagacaa acggcgaaac cggggagatc
     4141 gtgtgggata agggccggga ttttgccacc gtgcggaaag tgctgagcat gccccaagtg
     4201 aatatcgtga aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc
     4261 aagaggaaca gcgataagct gatcgccaga aagaaggact gggaccctaa gaagtacggc
     4321 ggcttcgaca gccccaccgt ggcctattct gtgctggtgg tggccaaagt ggaaaagggc
     4381 aagtccaaga aactgaagag tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc
     4441 agcttcgaga agaatcccat cgactttctg gaagccaagg gctacaaaga agtgaaaaag
     4501 gacctgatca tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg ccggaagaga
     4561 atgctggcct ctgccggcga actgcagaag ggaaacgaac tggccctgcc ctccaaatat
     4621 gtgaacttcc tgtacctggc cagccactat gagaagctga agggctcccc cgaggataat
     4681 gagcagaaac agctgtttgt ggaacagcac aagcactacc tggacgagat catcgagcag
     4741 atcagcgagt tctccaagag agtgatcctg gccgacgcta atctggacaa agtgctgtcc
     4801 gcctacaaca agcaccggga taagcccatc agagagcagg ccgagaatat catccacctg
     4861 tttaccctga ccaatctggg agcccctgcc gccttcaagt actttgacac caccatcgac
     4921 cggaagaggt acaccagcac caaagaggtg ctggacgcca ccctgatcca ccagagcatc
     4981 accggcctgt acgagacacg gatcgacctg tctcagctgg gaggcgacaa aaggccggcg
     5041 gccacgaaaa aggccggcca ggcaaaaaag aaaaagtaat tcctagagct cgctgatcag
     5101 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     5161 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     5221 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     5281 aggattggga agagaatagc aggcatgctg gggagcggcc gcaggaaccc ctagtgatgg
     5341 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
     5401 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc
     5461 aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
     5521 gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     5581 tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt
     5641 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     5701 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga
     5761 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     5821 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg
     5881 ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct
     5941 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg
     6001 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     6061 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     6121 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     6181 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     6241 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     6301 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     6361 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     6421 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     6481 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     6541 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     6601 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     6661 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     6721 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     6781 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     6841 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     6901 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     6961 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     7021 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     7081 agccggtgag cgtggaagcc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     7141 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     7201 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     7261 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     7321 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     7381 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     7441 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     7501 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     7561 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     7621 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     7681 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     7741 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     7801 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     7861 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     7921 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     7981 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     8041 ctggcctttt gctcacatgc tagaggtacc
//
