LOCUS       sequence-308957-       12922 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cas9 at very low level.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12922)
  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 12922)
  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(XL)Cas9
FEATURES             Location/Qualifiers
     source          1..12922
                     /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"
     primer_bind     complement(5278..5299)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5539..5558)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             5573..9676
                     /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             5573..9676
                     /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             5573..9676
                     /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             9689..9709
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          9796..9861
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             9991..10011
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    10283..10417
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10320..10339)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10374..10393
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    10446..10515
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     promoter        complement(10734..10752)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(10773..10789)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10773..10789)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10786..10808)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10797..10813
                     /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(10821..10827,10828..10845,10846..10851))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10866..10887
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11004..11021)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11175..11763)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11255..11274)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11934..12725,12726..12794))
                     /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     12557..12576
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12795..12899)
                     /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 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     5341 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     5401 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
     5461 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     5521 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgcctaata acatggacaa
     5581 gaagtactcc attgggctcg atatcggcac aaacagcgtc ggctgggccg tcattacgga
     5641 cgagtacaag gtgccgagca aaaaattcaa agttctgggc aataccgatc gccacagcat
     5701 aaagaagaac ctcattggcg ccctcctgtt cgactccggg gagacggccg aagccacgcg
     5761 gctcaaaaga acagcacggc gcagatatac ccgcagaaag aatcggatct gctacctgca
     5821 ggagatcttt agtaatgaga tggctaaggt ggatgactct ttcttccata ggctggagga
     5881 gtcctttttg gtggaggagg ataaaaagca cgagcgccac ccaatctttg gcaatatcgt
     5941 ggacgaggtg gcgtaccatg aaaagtaccc aaccatatat catctgagga agaagcttgt
     6001 agacagtact gataaggctg acttgcggtt gatctatctc gcgctggcgc atatgatcaa
     6061 atttcgggga cacttcctca tcgaggggga cctgaaccca gacaacagcg atgtcgacaa
     6121 actctttatc caactggttc agacttacaa tcagcttttc gaagagaacc cgatcaacgc
     6181 atccggagtt gacgccaaag caatcctgag cgctaggctg tccaaatccc ggcggctcga
     6241 aaacctcatc gcacagctcc ctggggagaa gaagaacggc ctgtttggta atcttatcgc
     6301 cctgtcactc gggctgaccc ccaactttaa atctaacttc gacctggccg aagatgccaa
     6361 gcttcaactg agcaaagaca cctacgatga tgatctcgac aatctgctgg cccagatcgg
     6421 cgaccagtac gcagaccttt ttttggcggc aaagaacctg tcagacgcca ttctgctgag
     6481 tgatattctg cgagtgaaca cggagatcac caaagctccg ctgagcgcta gtatgatcaa
     6541 gcgctatgat gagcaccacc aagacttgac tttgctgaag gcccttgtca gacagcaact
     6601 gcctgagaag tacaaggaaa ttttcttcga tcagtctaaa aatggctacg ccggatacat
     6661 tgacggcgga gcaagccagg aggaatttta caaatttatt aagcccatct tggaaaaaat
     6721 ggacggcacc gaggagctgc tggtaaagct taacagagaa gatctgttgc gcaaacagcg
     6781 cactttcgac aatggaagca tcccccacca gattcacctg ggcgaactgc acgctatcct
     6841 caggcggcaa gaggatttct accccttttt gaaagataac agggaaaaga ttgagaaaat
     6901 cctcacattt cggataccct actatgtagg ccccctcgcc cggggaaatt ccagattcgc
     6961 gtggatgact cgcaaatcag aagagaccat cactccctgg aacttcgagg aagtcgtgga
     7021 taagggggcc tctgcccagt ccttcatcga aaggatgact aactttgata aaaatctgcc
     7081 taacgaaaag gtgcttccta aacactctct gctgtacgag tacttcacag tttataacga
     7141 gctcaccaag gtcaaatacg tcacagaagg gatgagaaag ccagcattcc tgtctggaga
     7201 gcagaagaaa gctatcgtgg acctcctctt caagacgaac cggaaagtta ccgtgaaaca
     7261 gctcaaagaa gactatttca aaaagattga atgtttcgac tctgttgaaa tcagcggagt
     7321 ggaggatcgc ttcaacgcat ccctgggaac gtatcacgat ctcctgaaaa tcattaaaga
     7381 caaggacttc ctggacaatg aggagaacga ggacattctt gaggacattg tcctcaccct
     7441 tacgttgttt gaagataggg agatgattga agaacgcttg aaaacttacg ctcatctctt
     7501 cgacgacaaa gtcatgaaac agctcaagag gcgccgatat acaggatggg ggcggctgtc
     7561 aagaaaactg atcaatggga tccgagacaa gcagagtgga aagacaatcc tggattttct
     7621 taagtccgat ggatttgcca accggaactt catgcagttg atccatgatg actctctcac
     7681 ctttaaggag gacatccaga aagcacaagt ttctggccag ggggacagtc ttcacgagca
     7741 catcgctaat cttgcaggta gcccagctat caaaaaggga atactgcaga ccgttaaggt
     7801 cgtggatgaa ctcgtcaaag taatgggaag gcataagccc gagaatatcg ttatcgagat
     7861 ggcccgagag aaccaaacta cccagaaggg acagaagaac agtagggaaa ggatgaagag
     7921 gattgaagag ggtataaaag aactggggtc ccaaatcctt aaggaacacc cagttgaaaa
     7981 cacccagctt cagaatgaga agctctacct gtactacctg cagaacggca gggacatgta
     8041 cgtggatcag gaactggaca tcaatcggct ctccgactac gacgtggatc atatcgtgcc
     8101 ccagtctttt ctcaaagatg attctattga taataaagtg ttgacaagat ccgataaaaa
     8161 tagagggaag agtgataacg tcccctcaga agaagttgtc aagaaaatga aaaattattg
     8221 gcggcagctg ctgaacgcca aactgatcac acaacggaag ttcgataatc tgactaaggc
     8281 tgaacgaggt ggcctgtctg agttggataa agccggcttc atcaaaaggc agcttgttga
     8341 gacacgccag atcaccaagc acgtggccca aattctcgat tcacgcatga acaccaagta
     8401 cgatgaaaat gacaaactga ttcgagaggt gaaagttatt actctgaagt ctaagctggt
     8461 ctcagatttc agaaaggact ttcagtttta taaggtgaga gagatcaaca attaccacca
     8521 tgcgcatgat gcctacctga atgcagtggt aggcactgca cttatcaaaa aatatcccaa
     8581 gcttgaatct gaatttgttt acggagacta taaagtgtac gatgttagga aaatgatcgc
     8641 aaagtctgag caggaaatag gcaaggccac cgctaagtac ttcttttaca gcaatattat
     8701 gaattttttc aagaccgaga ttacactggc caatggagag attcggaagc gaccacttat
     8761 cgaaacaaac ggagaaacag gagaaatcgt gtgggacaag ggtagggatt tcgcgacagt
     8821 ccggaaggtc ctgtccatgc cgcaggtgaa catcgttaaa aagaccgaag tacagaccgg
     8881 aggcttctcc aaggaaagta tcctcccgaa aaggaacagc gacaagctga tcgcacgcaa
     8941 aaaagattgg gaccccaaga aatacggcgg attcgattct cctacagtcg cttacagtgt
     9001 actggttgtg gccaaagtgg agaaagggaa gtctaaaaaa ctcaaaagcg tcaaggaact
     9061 gctgggcatc acaatcatgg agcgatcaag cttcgaaaaa aaccccatcg actttctcga
     9121 ggcgaaagga tataaagagg tcaaaaaaga cctcatcatt aagcttccca agtactctct
     9181 ctttgagctt gaaaacggcc ggaaacgaat gctcgctagt gcgggcgagc tgcagaaagg
     9241 taacgagctg gcactgccct ctaaatacgt taatttcttg tatctggcca gccactatga
     9301 aaagctcaaa gggtctcccg aagataatga gcagaagcag ctgttcgtgg aacaacacaa
     9361 acactacctt gatgagatca tcgagcaaat aagcgaattc tccaaaagag tgatcctcgc
     9421 cgacgctaac ctcgataagg tgctttctgc ttacaataag cacagggata agcccatcag
     9481 ggagcaggca gaaaacatta tccacttgtt tactctgacc aacttgggcg cgcctgcagc
     9541 cttcaagtac ttcgacacca ccatagacag aaagcggtac acctctacaa aggaggtcct
     9601 ggacgccaca ctgattcatc agtcaattac ggggctctat gaaacaagaa tcgacctctc
     9661 tcagctcggt ggagacagca gggctgaccc caagaagaag aggaaggtgt gagctagcgg
     9721 atctttgtga aggaacctta cttctgtggt gtgacataat tggacaaact acctacagag
     9781 atttaaagct ctaaggtaaa tataaaattt ttaagtgtat aatgtgttaa actactgatt
     9841 ctaattgttt gtgtatttta gattccaacc tatggaactg atgaatggga gcagtggtgg
     9901 aatgccttta atgaggaaaa cctgttttgc tcagaagaaa tgccatctag tgatgatgag
     9961 gctactgctg actctcaaca ttctactcct ccaaaaaaga agagaaaggt agaagacccc
    10021 aaggactttc cttcagaatt gctaagtttt ttgagtcatg ctgtgtttag taatagaact
    10081 cttgcttgct ttgctattta caccacaaag gaaaaagctg cactgctata caagaaaatt
    10141 atggaaaaat atttgatgta tagtgccttg actagagatc ataatcagcc ataccacatt
    10201 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
    10261 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
    10321 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
    10381 gtccaaactc atcaatgtat cttatcatgt ctggatccac tagtgtcgac gatgtaggtc
    10441 acggtctcga agccgcggtg cgggtgccag ggcgtgccct tgggctcccc gggcgcgtac
    10501 tccacctcac ccatctggtc catcatgatg aacgggtcga ggtggcggta gttgatcccg
    10561 gcgaacgcgc ggcgcaccgg gaagccctcg ccctcgaaac cgctgggcgc ggtggtcacg
    10621 gtgagcacgg gacgtgcgac ggcgtcggcg ggtgcggata cgcggggcag cgtcagcggg
    10681 ttctcgacgg tcacggcggg catgtcgaca ctagttctag ccagcttttg ttccctttag
    10741 tgagggttaa tttcgagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
    10801 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
    10861 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
    10921 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
    10981 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
    11041 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
    11101 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
    11161 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
    11221 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
    11281 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
    11341 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
    11401 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
    11461 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
    11521 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
    11581 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
    11641 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
    11701 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
    11761 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
    11821 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
    11881 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
    11941 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
    12001 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
    12061 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
    12121 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
    12181 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
    12241 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
    12301 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
    12361 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
    12421 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
    12481 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
    12541 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
    12601 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
    12661 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
    12721 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
    12781 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
    12841 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
    12901 acatttcccc gaaaagtgcc ac
//
