LOCUS       sequence-275227-       11531 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cas9 in the germline of Drosophila.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11531)
  AUTHORS   Kondo S, Ueda R
  TITLE     Highly improved gene targeting by germline-specific Cas9 expression 
            in Drosophila.
  JOURNAL   Genetics. 2013 Nov;195(3):715-21. doi: 10.1534/genetics.113.156737. 
            Epub 2013 Sep 3.
  PUBMED    24002648
REFERENCE   2  (bases 1 to 11531)
  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"; doi: 
            "10.1534/genetics.113.156737"; journalName: "Genetics"; date: 
            "2013-11"; volume: "195"; issue: "3"; pages: "715-21"
COMMENT     Sequence Label: pBFv-nosP-Cas9
FEATURES             Location/Qualifiers
     source          1..11531
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1168..1184)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1168..1184)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     regulatory      1198..1207
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1204..5307
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             1204..5307
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             1204..5307
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             5320..5340
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(6379..6399)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6379..6397)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6418..6434)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6418..6434)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6431..6453)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6442..6458
                     /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(6466..6472,6473..6490,6491..6496))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6511..6532
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6649..6666)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6820..7408)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6900..6919)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7579..8370,8371..8439))
                     /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     8202..8221
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8440..8544)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(8570..9025)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8707..8728)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8919..8938
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9152..9174
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     9166..9183
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     9167..9183
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9193..9212
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        9193..9211
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    9419..9488
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     CDS             complement(join(9688..9805,9882..9975,10067..10673,
                     10728..10861,10922..11082,11156..11181))
                     /codon_start=1
                     /product="tryptophan oxygenase"
                     /label=vermilion
                     /note="selectable marker, required to synthesize the brown 
                     eye pigment in Drosophila"
                     /translation="MSCPYAGNGNDHDDSAVPLTTEVGKIYGEYLMLDKLLDAQCMLSE
                     EDKRPVHDEHLFIITHQAYELWFKQIIFEFDSIRDMLDAEVIDETKTLEIVKRLNRVVL
                     ILKLLVDQVPILETMTPLDFMDFRKYLAPASGFQSLQFRLIENKLGVLTEQRVRYNQKY
                     SDVFSDEEARNSIRNSEKDPSLLELVQRWLERTPGLEESGFNFWAKFQESVDRFLEAQV
                     QSAMEEPVEKAKNYRLMDIEKRREVYRSIFDPAVHDALVRRGDRRFSHRALQGAIMITF
                     YRDEPRFSQPHQLLTLLMDIDSLITKWRYNHVIMVQRMIGSQQLGTGGSSGYQYLRSTL
                     SDRYKVFLDLFNLSTFLIPREAIPPLDETIRKKLINKSV"
ORIGIN
        1 gttgtcgaag aactgcgaga agaagccggc cctgcgacct ggcgcactat aaaagcgggg
       61 ctgcggagta aagtcatgac ttaggccggt tgcgccatgg ggttttatga gccaaactct
      121 aacctgctgc tgctgcaggt tctgcgaggc ctgctggcac gtgaagagac aggcgcgatc
      181 ccgcacaagg gcagctattc tgtactgcaa ggagaaaaaa tcattgaaag cttcgaccgt
      241 tttaacctcg aaatatgcac atgtaaggac ggatgtgagc gaacgccagt gatgaccggg
      301 atcagaggta acctaccatg gtggggatta ggtgaccgtt cgcaggtagt ttgatcggag
      361 cgaatgttcg gggggtctgg cgtcagaggc tctaaacttt atgtaattcc tgccgcgaaa
      421 cacgcacgta tcaagcagtc agctgttctc ttcgttcagc gcgcgccggt gttgcaaaac
      481 gagcgctctt cgccggcggt ggctcgtgcg atagttcgtt ttgtcggtaa tccgatgttg
      541 ccgcgccgat atcatgtgat gttgtcacag tgcgcgaaat tcgaatggtg gtgtgcagtg
      601 attgtgttgt gacggcgagt ggcgcgtgtg ggtgcttagt tttgggagat gttttcgtat
      661 ttttttgttg ataactcagg ctttgttgct gtgttgtagt actattttcc attgcgcggt
      721 gtccagcttt taattagtgg cacatattct tagcaagtaa aaattatttt gcatactatt
      781 aaatttctta taaattattt tctaaaatta agtttacctt ttcaatttta ctaaaaatat
      841 cgatatattt attatcgctg gaaaactaca ttattccacc tctaagcaag aaccgttagt
      901 tggcgcgtag ctttaccaca aaattcctgg aattgccgta cgcttcgcag ttgtttcaag
      961 ttgtctaagg gacatacgat tttttttgcc tctgcgtcac gattttaacc caaaagcgag
     1021 tttagttaca tgtacattat tattagataa agaagtatcg cgaatacttc agttgaataa
     1081 actgtgcttg gtttttgggt gaggatttgt ggaaagtaga gtgcgcgata accgtaactt
     1141 tcgacccgga ttttcgccgg cgcgccggat ccactagttc tagaggatcg aacccttgcc
     1201 accatggaca agaagtactc cattgggctc gatatcggca caaacagcgt cggctgggcc
     1261 gtcattacgg acgagtacaa ggtgccgagc aaaaaattca aagttctggg caataccgat
     1321 cgccacagca taaagaagaa cctcattggc gccctcctgt tcgactccgg ggagacggcc
     1381 gaagccacgc ggctcaaaag aacagcacgg cgcagatata cccgcagaaa gaatcggatc
     1441 tgctacctgc aggagatctt tagtaatgag atggctaagg tggatgactc tttcttccat
     1501 aggctggagg agtccttttt ggtggaggag gataaaaagc acgagcgcca cccaatcttt
     1561 ggcaatatcg tggacgaggt ggcgtaccat gaaaagtacc caaccatata tcatctgagg
     1621 aagaagcttg tagacagtac tgataaggct gacttgcggt tgatctatct cgcgctggcg
     1681 catatgatca aatttcgggg acacttcctc atcgaggggg acctgaaccc agacaacagc
     1741 gatgtcgaca aactctttat ccaactggtt cagacttaca atcagctttt cgaagagaac
     1801 ccgatcaacg catccggagt tgacgccaaa gcaatcctga gcgctaggct gtccaaatcc
     1861 cggcggctcg aaaacctcat cgcacagctc cctggggaga agaagaacgg cctgtttggt
     1921 aatcttatcg ccctgtcact cgggctgacc cccaacttta aatctaactt cgacctggcc
     1981 gaagatgcca agcttcaact gagcaaagac acctacgatg atgatctcga caatctgctg
     2041 gcccagatcg gcgaccagta cgcagacctt tttttggcgg caaagaacct gtcagacgcc
     2101 attctgctga gtgatattct gcgagtgaac acggagatca ccaaagctcc gctgagcgct
     2161 agtatgatca agcgctatga tgagcaccac caagacttga ctttgctgaa ggcccttgtc
     2221 agacagcaac tgcctgagaa gtacaaggaa attttcttcg atcagtctaa aaatggctac
     2281 gccggataca ttgacggcgg agcaagccag gaggaatttt acaaatttat taagcccatc
     2341 ttggaaaaaa tggacggcac cgaggagctg ctggtaaagc ttaacagaga agatctgttg
     2401 cgcaaacagc gcactttcga caatggaagc atcccccacc agattcacct gggcgaactg
     2461 cacgctatcc tcaggcggca agaggatttc tacccctttt tgaaagataa cagggaaaag
     2521 attgagaaaa tcctcacatt tcggataccc tactatgtag gccccctcgc ccggggaaat
     2581 tccagattcg cgtggatgac tcgcaaatca gaagagacca tcactccctg gaacttcgag
     2641 gaagtcgtgg ataagggggc ctctgcccag tccttcatcg aaaggatgac taactttgat
     2701 aaaaatctgc ctaacgaaaa ggtgcttcct aaacactctc tgctgtacga gtacttcaca
     2761 gtttataacg agctcaccaa ggtcaaatac gtcacagaag ggatgagaaa gccagcattc
     2821 ctgtctggag agcagaagaa agctatcgtg gacctcctct tcaagacgaa ccggaaagtt
     2881 accgtgaaac agctcaaaga agactatttc aaaaagattg aatgtttcga ctctgttgaa
     2941 atcagcggag tggaggatcg cttcaacgca tccctgggaa cgtatcacga tctcctgaaa
     3001 atcattaaag acaaggactt cctggacaat gaggagaacg aggacattct tgaggacatt
     3061 gtcctcaccc ttacgttgtt tgaagatagg gagatgattg aagaacgctt gaaaacttac
     3121 gctcatctct tcgacgacaa agtcatgaaa cagctcaaga ggcgccgata tacaggatgg
     3181 gggcggctgt caagaaaact gatcaatggg atccgagaca agcagagtgg aaagacaatc
     3241 ctggattttc ttaagtccga tggatttgcc aaccggaact tcatgcagtt gatccatgat
     3301 gactctctca cctttaagga ggacatccag aaagcacaag tttctggcca gggggacagt
     3361 cttcacgagc acatcgctaa tcttgcaggt agcccagcta tcaaaaaggg aatactgcag
     3421 accgttaagg tcgtggatga actcgtcaaa gtaatgggaa ggcataagcc cgagaatatc
     3481 gttatcgaga tggcccgaga gaaccaaact acccagaagg gacagaagaa cagtagggaa
     3541 aggatgaaga ggattgaaga gggtataaaa gaactggggt cccaaatcct taaggaacac
     3601 ccagttgaaa acacccagct tcagaatgag aagctctacc tgtactacct gcagaacggc
     3661 agggacatgt acgtggatca ggaactggac atcaatcggc tctccgacta cgacgtggat
     3721 catatcgtgc cccagtcttt tctcaaagat gattctattg ataataaagt gttgacaaga
     3781 tccgataaaa atagagggaa gagtgataac gtcccctcag aagaagttgt caagaaaatg
     3841 aaaaattatt ggcggcagct gctgaacgcc aaactgatca cacaacggaa gttcgataat
     3901 ctgactaagg ctgaacgagg tggcctgtct gagttggata aagccggctt catcaaaagg
     3961 cagcttgttg agacacgcca gatcaccaag cacgtggccc aaattctcga ttcacgcatg
     4021 aacaccaagt acgatgaaaa tgacaaactg attcgagagg tgaaagttat tactctgaag
     4081 tctaagctgg tctcagattt cagaaaggac tttcagtttt ataaggtgag agagatcaac
     4141 aattaccacc atgcgcatga tgcctacctg aatgcagtgg taggcactgc acttatcaaa
     4201 aaatatccca agcttgaatc tgaatttgtt tacggagact ataaagtgta cgatgttagg
     4261 aaaatgatcg caaagtctga gcaggaaata ggcaaggcca ccgctaagta cttcttttac
     4321 agcaatatta tgaatttttt caagaccgag attacactgg ccaatggaga gattcggaag
     4381 cgaccactta tcgaaacaaa cggagaaaca ggagaaatcg tgtgggacaa gggtagggat
     4441 ttcgcgacag tccggaaggt cctgtccatg ccgcaggtga acatcgttaa aaagaccgaa
     4501 gtacagaccg gaggcttctc caaggaaagt atcctcccga aaaggaacag cgacaagctg
     4561 atcgcacgca aaaaagattg ggaccccaag aaatacggcg gattcgattc tcctacagtc
     4621 gcttacagtg tactggttgt ggccaaagtg gagaaaggga agtctaaaaa actcaaaagc
     4681 gtcaaggaac tgctgggcat cacaatcatg gagcgatcaa gcttcgaaaa aaaccccatc
     4741 gactttctcg aggcgaaagg atataaagag gtcaaaaaag acctcatcat taagcttccc
     4801 aagtactctc tctttgagct tgaaaacggc cggaaacgaa tgctcgctag tgcgggcgag
     4861 ctgcagaaag gtaacgagct ggcactgccc tctaaatacg ttaatttctt gtatctggcc
     4921 agccactatg aaaagctcaa agggtctccc gaagataatg agcagaagca gctgttcgtg
     4981 gaacaacaca aacactacct tgatgagatc atcgagcaaa taagcgaatt ctccaaaaga
     5041 gtgatcctcg ccgacgctaa cctcgataag gtgctttctg cttacaataa gcacagggat
     5101 aagcccatca gggagcaggc agaaaacatt atccacttgt ttactctgac caacttgggc
     5161 gcgcctgcag ccttcaagta cttcgacacc accatagaca gaaagcggta cacctctaca
     5221 aaggaggtcc tggacgccac actgattcat cagtcaatta cggggctcta tgaaacaaga
     5281 atcgacctct ctcagctcgg tggagacagc agggctgacc ccaagaagaa gaggaaggtg
     5341 tgaaagggtt cgatccctac cggtgagagg gcgaatccag ctctggagca gaggctctgg
     5401 cagcttttgc agcgtttata taacatgaaa tatatatacg cattccgatc aaagctgggt
     5461 taaccagata gatagatagt aacgtttaaa tagcgcctgg cgcgttcgat tttaaagaga
     5521 tttagagcgt tatcccgtgc ctatagatct tatagtatag acaacgaacg atcactcaaa
     5581 tccaagtcaa taattcaaga atttatgtct gtttctgtga aagggaaact aattttgtta
     5641 aagaagactt acaatatcgt aatacttgtt caatcgtcgt ggccgataga aatatcttac
     5701 aatccgaaag ttgatgaatg gaattggtct gcaactggtc gccttcattt cgtaaaatgt
     5761 tcgcttgcgg ccgaaaaatt tcgatatatc tacaattgat ctacaatctt tactaaattt
     5821 tgaaaaagga acactttgaa tttcgaactg tcaatcgtat cattagaatt taatctaaat
     5881 ttaaatcttg ctaaaggaaa tagcaaggaa cactttcgtc gtcggctacg cattcattgt
     5941 aaaattttaa attttgacat tccgcacttt ttgatagata agcgaagagt atttttatta
     6001 catgtatcgc aagtattcat ttcaacacac atatctatat atatatatat atatatatgt
     6061 tatatattta ttcaattttg tttaccattg atcaattttt cacacatgaa acaaccgcca
     6121 gcattatata atttttttat ttttttaaaa aatgtgtaca catattctga aaatgaaaaa
     6181 ttcaatggct cgagtgccaa ataaagaaat ggttacaatt taaggaaaca aatgtccttc
     6241 ttgcgtttga aacaactaat ccttttcgcc ctcgcggcgt ttctcgaaaa gggccaggaa
     6301 gatgccatcg catgcacttg gctttccacc gttggtatcg attctctggg acgatgagtc
     6361 gagctccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc
     6421 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     6481 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     6541 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     6601 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     6661 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     6721 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     6781 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     6841 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     6901 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     6961 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     7021 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     7081 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     7141 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     7201 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     7261 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     7321 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     7381 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     7441 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     7501 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     7561 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     7621 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     7681 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     7741 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     7801 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     7861 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     7921 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     7981 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     8041 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     8101 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     8161 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     8221 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     8281 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     8341 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     8401 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     8461 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     8521 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc
     8581 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     8641 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     8701 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     8761 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     8821 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     8881 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     8941 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     9001 cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa
     9061 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
     9121 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
     9181 agtgagcgcg cgtaatacga ctcactatag ggcgaattgg gtacctcgac atgcccgccg
     9241 tgaccgtcga gaacccgctg acgctgcccc gcgtatccgc acccgccgac gccgtcgcac
     9301 gtcccgtgct caccgtgacc accgcgccca gcggtttcga gggcgagggc ttcccggtgc
     9361 gccgcgcgtt cgccgggatc aactaccgcc acctcgaccc gttcatcatg atggaccaga
     9421 tgggtgaggt ggagtacgcg cccggggagc ccaagggcac gccctggcac ccgcaccgcg
     9481 gcttcgagac cgtgacctac atcgtcgaca agcttggatt tattttgtta tgttatatgt
     9541 attatatgtc agacataaag aaaaggaaca catcaaatgt gataacaaag actaaacaag
     9601 taattttatt acaccaaaac gacaaaacag taggcagaac aaacaacgca tagccaaaca
     9661 ttgacgaatt ggataccctg ccgattgtca gacacttttg ttgatcagtt tcttgcgaat
     9721 ggtctcgtcc agcggtggaa tcgcctcgcg gggaatcaga aaagtggaca gattgaacag
     9781 atccagaaac accttgtacc gatcactgaa accaaaaaaa aacaaaggga gaacagtttg
     9841 agttcattga tccccgatat aatcacatct gcgatgatca cctgagagtg gagcgcagat
     9901 attgatatcc agacgagcca ccagtgccca actgttggga tccaatcatg cgttgcacca
     9961 tgatcacgtg attgtctgcg gcgggaatag aaagtatttg gttaggaaaa ccagtcttaa
    10021 acataagata tatttataaa agagtatcaa agaatgcaat acttacatct ccacttggtt
    10081 attaacgagt cgatgtccat gagcagggtg agcaactggt gtggttggct gaacctgggt
    10141 tcatccctat agaaggtgat catgatggct ccctgaaggg cacgatggct aaaccggcga
    10201 tccccacgac gcaccagtgc atcgtgcact gccggatcaa agatggagcg atacacctcg
    10261 cgtcgcttct caatgtccat gaggcggtag tttttcgcct tctccacggg ctcctccatg
    10321 gcgctctgta cctgcgcctc caggaatcga tcgacgctct cctgaaactt ggcccagaag
    10381 ttgaagccac tctcctccag tccgggcgtc ctctccagcc atcgctgcac tagctccagt
    10441 agcgagggat ctttctccga gttgcgaatc gagttccgcg cctcctcgtc gctaaagaca
    10501 tccgagtact tctggttgta tctcacccgc tgctctgtca gaactcccag cttgttctcg
    10561 atcaaacgga actgcagcga ctgaaaacca gatgcgggtg ccaggtactt gcggaagtcc
    10621 atgaagtcta gcggggtcat ggtctccaga atgggcactt ggtccaccag gagctgtaca
    10681 aaggaagtta taaacggatt ttggtaagag attcagaaag cactcacttt tagaatcaga
    10741 accactcggt tcagtcgctt gacaatctcc agcgtcttgg tttcatcgat gacctctgca
    10801 tccaacatgt ctcgtatgga gtcgaactca aagatgatct gcttgaacca aagctcgtag
    10861 gctgtggcga aggtacttaa atgccattga gtgttgtcat caaagttgta aacctactca
    10921 ccctggtgcg tgatgatgaa cagatgctca tcgtgcacgg gtcgcttgtc ctcctcggac
    10981 agcatacact gggcatccag cagtttgtcc agcatcagat actctccata gattttgccc
    11041 acttccgtgg ttaatggcac cgccgaatca tcgtgatcgt ttctgtatgg gtttgaattg
    11101 aatcgcagaa ctgaagatcg attggcattc ctggacagca cgtgctggtg ctcacccgtt
    11161 tcctgcatag ggacagctca tggtgcacag ctcagatcag atcgtgactc ctcgagcggc
    11221 ggatgctggc gaactgatct ccgccagcgg accggagatg agaccccagc gaaccgataa
    11281 cagagcgaga gagctccagt tccgactgat tgcacagtcg gtgatctggg cgatgggcac
    11341 tgccagatag gctgggaatt atcaatcact tgaggtgaaa gtgcggcgca cacaaatcca
    11401 agcttgatat cgaattccct gcagggccgc gccgatttca gggcatccga ttcttccagc
    11461 ttctgggcct cctcacggaa ctttcggata ttgtccttga tctccttgtt cttatccatc
    11521 tccgccttca t
//
