LOCUS       sequence-308953-       12703 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cas9 at medium level.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12703)
  AUTHORS   Port F, Strein C, Stricker M, Rauscher B, Heigwer F, Zhou J, 
            Beyersdorffer C, Frei J, Hess A, Kern K, Lange L, Langner N, Malamud
            R, Pavlovic B, Radecke K, Schmitt L, Voos L, Valentini E, Boutros M
  TITLE     A large-scale resource for tissue-specific CRISPR mutagenesis in 
            Drosophila.
  JOURNAL   Elife. 2020 Feb 13;9. pii: 53865. doi: 10.7554/eLife.53865.
  PUBMED    32053108
REFERENCE   2  (bases 1 to 12703)
  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: "Elife."; 
            date: "2020-02-13"; volume: "9. pii: 53865. doi"; pages: " 
            10.7554/eLife.53865"
COMMENT     Sequence Label: UAS-u(M)Cas9
FEATURES             Location/Qualifiers
     source          1..12703
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     623..642
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        623..641
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(675..693)
                     /label=pCasper-F
                     /note="Drosophila mini-white gene, reverse primer"
     CDS             join(1145..1216,1618..1891,1966..2620,2682..2997,3201..3332,
                     3403..4017)
                     /codon_start=1
                     /gene="white"
                     /product="Drosophila white gene eye color pigment"
                     /label=mini-white
                     /note="This is a modified version of the white gene lacking
                     part of the first intron."
                     /translation="MGQEDQELLIRGGSKHPSAEHLNNGDSGAASQSCINQGFGQAKNY
                     GTLRPPSPPEDSGSGSGQLAENLTYAWHNMDIFGAVNQPGSGWRQLVNRTRGLFCNERH
                     IPAPRKHLLKNVCGVAYPGELLAVMGSSGAGKTTLLNALAFRSPQGIQVSPSGMRLLNG
                     QPVDAKEMQARCAYVQQDDLFIGSLTAREHLIFQAMVRMPRHLTYRQRVARVDQVIQEL
                     SLSKCQHTIIGVPGRVKGLSGGERKRLAFASEALTDPPLLICDEPTSGLDSFTAHSVVQ
                     VLKKLSQKGKTVILTIHQPSSELFELFDKILLMAEGRVAFLGTPSEAVDFFSYVGAQCP
                     TNYNPADFYVQVLAVVPGREIESRDRIAKICDNFAISKVARDMEQLLATKNLEKPLEQP
                     ENGYTYKATWFMQFRAVLWRSWLSVLKEPLLVKVRLIQTTMVAILIGLIFLGQQLTQVG
                     VMNINGAIFLFLTNMTFQNVFATINVFTSELPVFMREARSRLYRCDTYFLGKTIAELPL
                     FLTVPLVFTAIAYPMIGLRAGVLHFFNCLALVTLVANVSTSFGYLISCASSSTSMALSV
                     GPPVIIPFLLFGGFFLNSGSVPVYLKWLSYLSWFRYANEGLLINQWADVEPGEISCTSS
                     NTTCPSSGKVILETLNFSAADLPLDYVGLAILIVSFRVLAYLALRLRARRKE"
     protein_bind    4792..4825
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    4858..4952
                     /label=5X UAS
                     /bound_moiety="GAL4"
                     /note="five tandem copies of the ""ScaI site"" 17-mer 
                     CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) 
                     that efficiently binds yeast Gal4 (Webster et al., 1988; 
                     Pfeiffer et al., 2010)"
     promoter        4971..5209
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     primer_bind     5156..5177
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     CDS             5354..9457
                     /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             5354..9457
                     /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             5354..9457
                     /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             9470..9490
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          9577..9642
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             9772..9792
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    10064..10198
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10101..10120)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10155..10174
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    10227..10296
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     promoter        complement(10515..10533)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(10554..10570)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10554..10570)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10567..10589)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10578..10594
                     /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(10602..10608,10609..10626,10627..10632))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10647..10668
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10785..10802)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10956..11544)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11036..11055)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11715..12506,12507..12575))
                     /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     12338..12357
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12576..12680)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg gtacgtaccg
      661 ggcccctagt atgtatgtaa gttaataaaa cccatttttg cggaaagtag ataaaaaaaa
      721 catttttttt ttttactgca ctggatatca ttgaacttat ctgatcagtt ttaaatttac
      781 ttcgatccaa gggtatttga tgtaccaggt tctttcgatt acctctcact caaaatgaca
      841 ttccactcaa agtcagcgct gtttgcctcc ttctctgtcc acagaaatat cgccgtctct
      901 ttcgccgctg cgtccgctat ctctttcgcc accgtttgta gcgttacgta gcgtcaatgt
      961 ccgccttcag ttgcattttg tcagcggttt cgtgacgaag ctccaagcgg tttacgccat
     1021 caattaaaca caaagtgctg tgccaaaact cctctcgctt cttatttttg tttgtttttt
     1081 gagtgattgg ggtggtgatt ggttttgggt gggtaagcag gggaaagtgt gaaaaatccc
     1141 ggcaatgggc caagaggatc aggagctatt aattcgcgga ggcagcaaac acccatctgc
     1201 cgagcatctg aacaatgtga gtagtacatg tgcatacatc ttaagttcac ttgatctata
     1261 ggaactgcga ttgcaacatc aaattgtctg cggcgtgaga actgcgaccc acaaaaatcc
     1321 caaaccgcaa ttgcacaaac aaatagtgac acgaaacaga ttattctggt agctgttctc
     1381 gctatataag acaatttttg agatcatatc atgatcaaga catctaaagg cattcatttt
     1441 cgactatatt cttttttaca aaaaatataa caaccagata ttttaagctg atcctagatg
     1501 cacaaaaaat aaataaaagt ataaacctac ttcgtaggat acttcggggt actttttgtt
     1561 ctgggttaga tgagcataac gcttgtagtt gatatttgag atcccctatc attgcagggt
     1621 gacagcggag cggcttcgca gagctgcatt aaccagggct tcgggcaggc caaaaactac
     1681 ggcacgctcc ggccacccag tccgccggag gactccggtt cagggagcgg ccaactagcc
     1741 gagaacctca cctatgcctg gcacaatatg gacatctttg gggcggtcaa tcagccgggc
     1801 tccggatggc ggcagctggt caaccggaca cgcggactat tctgcaacga gcgacacata
     1861 ccggcgccca ggaaacattt gctcaagaac ggtgagtttc tattcgcagt cggctgatct
     1921 gtgtgaaatc ttaataaagg gtccaattac caatttgaaa ctcagtttgc ggcgtggcct
     1981 atccgggcga acttttggcc gtgatgggca gttccggtgc cggaaagacg accctgctga
     2041 atgcccttgc ctttcgatcg ccgcagggca tccaagtatc gccatccggg atgcgactgc
     2101 tcaatggcca acctgtggac gccaaggaga tgcaggccag gtgcgcctat gtccagcagg
     2161 atgacctctt tatcggctcc ctaacggcca gggaacacct gattttccaa gccatggtgc
     2221 ggatgccacg acatctgacc tatcggcagc gagtggcccg cgtggatcag gtgatccagg
     2281 agctttcgct cagcaaatgt cagcacacga tcatcggtgt gcccggcagg gtgaaaggtc
     2341 tgtccggcgg agaaaggaag cgtctggcat tcgcctccga ggctctaacc gatccgccgc
     2401 ttctgatctg cgatgagccc acctccggac tggactcctt taccgcccac agcgtcgtcc
     2461 aggtgctgaa gaagctgtcg cagaagggca agaccgtcat cctgaccatt catcagccgt
     2521 cttccgagct gtttgagctc tttgacaaga tccttctgat ggccgagggc agggtagctt
     2581 tcttgggcac tcccagcgaa gccgtcgact tcttttccta gtgagttcga tgtgtttatt
     2641 aagggtatct agtattacat aacatctcaa ctcctatcca gcgtgggtgc ccagtgtcct
     2701 accaactaca atccggcgga cttttacgta caggtgttgg ccgttgtgcc cggacgggag
     2761 atcgagtccc gtgatcggat cgccaagata tgcgacaatt ttgccattag caaagtagcc
     2821 cgggatatgg agcagttgtt ggccaccaaa aatctggaga agccactgga gcagccggag
     2881 aatgggtaca cctacaaggc cacctggttc atgcagttcc gggcggtcct gtggcgatcc
     2941 tggctgtcgg tgctcaagga accactcctc gtaaaagtgc gacttattca gacaacggtg
     3001 agtggttcca gtggaaacaa atgatataac gcttacaatt cttggaaaca aattcgctag
     3061 attttagata gaattgcctg attccacacc cttcttagtt tttttcaatg agatgtatag
     3121 tttatagttt tgcagaagat aaataaattt catttaactc gcgaatatta atgagatgcg
     3181 agtaacattt taatttgcag atggttgcca tcttgattgg cctcatcttt ttgggccaac
     3241 aactcacgca agtgggtgtg atgaatatca acggagccat cttcctcttc ctgaccaaca
     3301 tgacctttca aaacgtcttt gccacgataa atgtaagtca tgtttagaat acatttgcat
     3361 ttcaataatt tactaacttt ctaatgaatc gattcgattt aggtgttcac ctcagagctg
     3421 ccagttttta tgagggaggc ccgaagtcga ctttatcgct gtgacacata ctttctgggc
     3481 aaaacgattg ccgaattgcc gctttttctc acagtgccac tggtcttcac ggcgattgcc
     3541 tatccgatga tcggactgcg ggccggagtg ctgcacttct tcaactgcct ggcgctggtc
     3601 actctggtgg ccaatgtgtc aacgtccttc ggatatctaa tatcctgcgc cagctcctcg
     3661 acctcgatgg cgctgtctgt gggtccgccg gttatcatac cattcctgct ctttggcggc
     3721 ttcttcttga actcgggctc ggtgccagta tacctcaaat ggttgtcgta cctctcatgg
     3781 ttccgttacg ccaacgaggg tctgctgatt aaccaatggg cggacgtgga gccgggcgaa
     3841 attagctgca catcgtcgaa caccacgtgc cccagttcgg gcaaggtcat cctggagacg
     3901 cttaacttct ccgccgccga tctgccgctg gactacgtgg gtctggccat tctcatcgtg
     3961 agcttccggg tgctcgcata tctggctcta agacttcggg cccgacgcaa ggagtagccg
     4021 acatatatcc gaaataactg cttgtttttt ttttttacca ttattaccat cgtgtttact
     4081 gtttattgcc ccctcaaaaa gctaatgtaa ttatatttgt gccaataaaa acaagatatg
     4141 acctatagaa tacaagtatt tccccttcga acatccccac aagtagactt tggatttgtc
     4201 ttctaaccaa aagacttaca cacctgcata ccttacatca aaaactcgtt tatcgctaca
     4261 taaaacaccg ggatatattt tttatataca tacttttcaa atcgcgcgcc ctcttcataa
     4321 ttcacctcca ccacaccacg tttcgtagtt gctctttcgc tgtctcccac ccgctctccg
     4381 caacacattc accttttgtt cgacgacctt ggagcgactg tcgttagttc cgcgcgattc
     4441 ggttcgctca aatggttccg agtggttcat ttcgtctcaa tagaaattag taataaatat
     4501 ttgtatgtac aatttatttg ctccaatata tttgtatata tttccctcac agctatattt
     4561 attctaattt aatattatga ctttttaagg taattttttg tgacctgttc ggagtgatta
     4621 gcgttacaat ttgaactgaa agtgacatcc agtgtttgtt ccttgtgtag atgcatctca
     4681 aaaaaatggt gggcataata gtgttgttta tatatatcaa aaataacaac tataataata
     4741 agaatacatt taatttagaa aatgcttgga tttcactgga actaggctag cataacttcg
     4801 tataatgtat gctatacgaa gttatgctag cggatccaag cttgcatgcc tgcaggtcgg
     4861 agtactgtcc tccgagcgga gtactgtcct ccgagcggag tactgtcctc cgagcggagt
     4921 actgtcctcc gagcggagta ctgtcctccg agcggagact ctagcgagcg ccggagtata
     4981 aatagaggcg cttcgtctac ggagcgacaa ttcaattcaa acaagcaaag tgaacacgtc
     5041 gctaagcgaa agctaagcaa ataaacaagc gcagctgaac aagctaaaca atctgcagta
     5101 aagtgcaagt taaagtgaat caattaaaag taaccagcaa ccaagtaaat caactgcaac
     5161 tactgaaatc tgccaagaag taattattga atacaagaag agaactctga atagggaatt
     5221 gggaattcca acatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     5281 gtcgagctgg acggctctag agccctcgcc ggacacgctg aacttgtggc cgtttacgtc
     5341 gctagataat aacatggaca agaagtactc cattgggctc gatatcggca caaacagcgt
     5401 cggctgggcc gtcattacgg acgagtacaa ggtgccgagc aaaaaattca aagttctggg
     5461 caataccgat cgccacagca taaagaagaa cctcattggc gccctcctgt tcgactccgg
     5521 ggagacggcc gaagccacgc ggctcaaaag aacagcacgg cgcagatata cccgcagaaa
     5581 gaatcggatc tgctacctgc aggagatctt tagtaatgag atggctaagg tggatgactc
     5641 tttcttccat aggctggagg agtccttttt ggtggaggag gataaaaagc acgagcgcca
     5701 cccaatcttt ggcaatatcg tggacgaggt ggcgtaccat gaaaagtacc caaccatata
     5761 tcatctgagg aagaagcttg tagacagtac tgataaggct gacttgcggt tgatctatct
     5821 cgcgctggcg catatgatca aatttcgggg acacttcctc atcgaggggg acctgaaccc
     5881 agacaacagc gatgtcgaca aactctttat ccaactggtt cagacttaca atcagctttt
     5941 cgaagagaac ccgatcaacg catccggagt tgacgccaaa gcaatcctga gcgctaggct
     6001 gtccaaatcc cggcggctcg aaaacctcat cgcacagctc cctggggaga agaagaacgg
     6061 cctgtttggt aatcttatcg ccctgtcact cgggctgacc cccaacttta aatctaactt
     6121 cgacctggcc gaagatgcca agcttcaact gagcaaagac acctacgatg atgatctcga
     6181 caatctgctg gcccagatcg gcgaccagta cgcagacctt tttttggcgg caaagaacct
     6241 gtcagacgcc attctgctga gtgatattct gcgagtgaac acggagatca ccaaagctcc
     6301 gctgagcgct agtatgatca agcgctatga tgagcaccac caagacttga ctttgctgaa
     6361 ggcccttgtc agacagcaac tgcctgagaa gtacaaggaa attttcttcg atcagtctaa
     6421 aaatggctac gccggataca ttgacggcgg agcaagccag gaggaatttt acaaatttat
     6481 taagcccatc ttggaaaaaa tggacggcac cgaggagctg ctggtaaagc ttaacagaga
     6541 agatctgttg cgcaaacagc gcactttcga caatggaagc atcccccacc agattcacct
     6601 gggcgaactg cacgctatcc tcaggcggca agaggatttc tacccctttt tgaaagataa
     6661 cagggaaaag attgagaaaa tcctcacatt tcggataccc tactatgtag gccccctcgc
     6721 ccggggaaat tccagattcg cgtggatgac tcgcaaatca gaagagacca tcactccctg
     6781 gaacttcgag gaagtcgtgg ataagggggc ctctgcccag tccttcatcg aaaggatgac
     6841 taactttgat aaaaatctgc ctaacgaaaa ggtgcttcct aaacactctc tgctgtacga
     6901 gtacttcaca gtttataacg agctcaccaa ggtcaaatac gtcacagaag ggatgagaaa
     6961 gccagcattc ctgtctggag agcagaagaa agctatcgtg gacctcctct tcaagacgaa
     7021 ccggaaagtt accgtgaaac agctcaaaga agactatttc aaaaagattg aatgtttcga
     7081 ctctgttgaa atcagcggag tggaggatcg cttcaacgca tccctgggaa cgtatcacga
     7141 tctcctgaaa atcattaaag acaaggactt cctggacaat gaggagaacg aggacattct
     7201 tgaggacatt gtcctcaccc ttacgttgtt tgaagatagg gagatgattg aagaacgctt
     7261 gaaaacttac gctcatctct tcgacgacaa agtcatgaaa cagctcaaga ggcgccgata
     7321 tacaggatgg gggcggctgt caagaaaact gatcaatggg atccgagaca agcagagtgg
     7381 aaagacaatc ctggattttc ttaagtccga tggatttgcc aaccggaact tcatgcagtt
     7441 gatccatgat gactctctca cctttaagga ggacatccag aaagcacaag tttctggcca
     7501 gggggacagt cttcacgagc acatcgctaa tcttgcaggt agcccagcta tcaaaaaggg
     7561 aatactgcag accgttaagg tcgtggatga actcgtcaaa gtaatgggaa ggcataagcc
     7621 cgagaatatc gttatcgaga tggcccgaga gaaccaaact acccagaagg gacagaagaa
     7681 cagtagggaa aggatgaaga ggattgaaga gggtataaaa gaactggggt cccaaatcct
     7741 taaggaacac ccagttgaaa acacccagct tcagaatgag aagctctacc tgtactacct
     7801 gcagaacggc agggacatgt acgtggatca ggaactggac atcaatcggc tctccgacta
     7861 cgacgtggat catatcgtgc cccagtcttt tctcaaagat gattctattg ataataaagt
     7921 gttgacaaga tccgataaaa atagagggaa gagtgataac gtcccctcag aagaagttgt
     7981 caagaaaatg aaaaattatt ggcggcagct gctgaacgcc aaactgatca cacaacggaa
     8041 gttcgataat ctgactaagg ctgaacgagg tggcctgtct gagttggata aagccggctt
     8101 catcaaaagg cagcttgttg agacacgcca gatcaccaag cacgtggccc aaattctcga
     8161 ttcacgcatg aacaccaagt acgatgaaaa tgacaaactg attcgagagg tgaaagttat
     8221 tactctgaag tctaagctgg tctcagattt cagaaaggac tttcagtttt ataaggtgag
     8281 agagatcaac aattaccacc atgcgcatga tgcctacctg aatgcagtgg taggcactgc
     8341 acttatcaaa aaatatccca agcttgaatc tgaatttgtt tacggagact ataaagtgta
     8401 cgatgttagg aaaatgatcg caaagtctga gcaggaaata ggcaaggcca ccgctaagta
     8461 cttcttttac agcaatatta tgaatttttt caagaccgag attacactgg ccaatggaga
     8521 gattcggaag cgaccactta tcgaaacaaa cggagaaaca ggagaaatcg tgtgggacaa
     8581 gggtagggat ttcgcgacag tccggaaggt cctgtccatg ccgcaggtga acatcgttaa
     8641 aaagaccgaa gtacagaccg gaggcttctc caaggaaagt atcctcccga aaaggaacag
     8701 cgacaagctg atcgcacgca aaaaagattg ggaccccaag aaatacggcg gattcgattc
     8761 tcctacagtc gcttacagtg tactggttgt ggccaaagtg gagaaaggga agtctaaaaa
     8821 actcaaaagc gtcaaggaac tgctgggcat cacaatcatg gagcgatcaa gcttcgaaaa
     8881 aaaccccatc gactttctcg aggcgaaagg atataaagag gtcaaaaaag acctcatcat
     8941 taagcttccc aagtactctc tctttgagct tgaaaacggc cggaaacgaa tgctcgctag
     9001 tgcgggcgag ctgcagaaag gtaacgagct ggcactgccc tctaaatacg ttaatttctt
     9061 gtatctggcc agccactatg aaaagctcaa agggtctccc gaagataatg agcagaagca
     9121 gctgttcgtg gaacaacaca aacactacct tgatgagatc atcgagcaaa taagcgaatt
     9181 ctccaaaaga gtgatcctcg ccgacgctaa cctcgataag gtgctttctg cttacaataa
     9241 gcacagggat aagcccatca gggagcaggc agaaaacatt atccacttgt ttactctgac
     9301 caacttgggc gcgcctgcag ccttcaagta cttcgacacc accatagaca gaaagcggta
     9361 cacctctaca aaggaggtcc tggacgccac actgattcat cagtcaatta cggggctcta
     9421 tgaaacaaga atcgacctct ctcagctcgg tggagacagc agggctgacc ccaagaagaa
     9481 gaggaaggtg tgagctagcg gatctttgtg aaggaacctt acttctgtgg tgtgacataa
     9541 ttggacaaac tacctacaga gatttaaagc tctaaggtaa atataaaatt tttaagtgta
     9601 taatgtgtta aactactgat tctaattgtt tgtgtatttt agattccaac ctatggaact
     9661 gatgaatggg agcagtggtg gaatgccttt aatgaggaaa acctgttttg ctcagaagaa
     9721 atgccatcta gtgatgatga ggctactgct gactctcaac attctactcc tccaaaaaag
     9781 aagagaaagg tagaagaccc caaggacttt ccttcagaat tgctaagttt tttgagtcat
     9841 gctgtgttta gtaatagaac tcttgcttgc tttgctattt acaccacaaa ggaaaaagct
     9901 gcactgctat acaagaaaat tatggaaaaa tatttgatgt atagtgcctt gactagagat
     9961 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
    10021 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
    10081 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
    10141 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatcca
    10201 ctagtgtcga cgatgtaggt cacggtctcg aagccgcggt gcgggtgcca gggcgtgccc
    10261 ttgggctccc cgggcgcgta ctccacctca cccatctggt ccatcatgat gaacgggtcg
    10321 aggtggcggt agttgatccc ggcgaacgcg cggcgcaccg ggaagccctc gccctcgaaa
    10381 ccgctgggcg cggtggtcac ggtgagcacg ggacgtgcga cggcgtcggc gggtgcggat
    10441 acgcggggca gcgtcagcgg gttctcgacg gtcacggcgg gcatgtcgac actagttcta
    10501 gccagctttt gttcccttta gtgagggtta atttcgagct tggcgtaatc atggtcatag
    10561 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
    10621 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
    10681 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
    10741 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
    10801 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
    10861 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
    10921 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
    10981 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
    11041 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
    11101 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
    11161 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
    11221 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
    11281 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
    11341 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
    11401 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
    11461 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
    11521 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
    11581 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
    11641 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
    11701 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
    11761 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
    11821 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
    11881 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
    11941 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
    12001 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
    12061 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
    12121 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
    12181 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
    12241 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
    12301 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
    12361 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
    12421 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
    12481 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
    12541 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
    12601 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
    12661 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cac
//
