LOCUS       sequence-272855-       13341 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 13341)
  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 13341)
  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(XXL)Cas9
FEATURES             Location/Qualifiers
     source          1..13341
                     /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"
     gene            666..4802
                     /gene="white"
                     /label=mini-white
                     /note="This modified version of the white gene lacks part 
                     of the first intron."
     primer_bind     complement(675..693)
                     /label=pCasper-F
                     /note="Drosophila mini-white gene, reverse primer"
     CDS             join(1145..1216,1618..1891,1966..2620,2682..2997,3218..3349,
                     3420..4034)
                     /codon_start=1
                     /gene="white"
                     /product="Drosophila white gene eye color pigment"
                     /label=mini-white
                     /note="This modified version of the white gene lacks part 
                     of the first intron."
                     /translation="MGQEDQELLIRGGSKHPSAEHLNNGDSGAASQSCINQGFGQAKNY
                     GTLRPPSPPEDSGSGSGQLAENLTYAWHNMDIFGAVNQPGSGWRQLVNRTRGLFCNERH
                     IPAPRKHLLKNVCGVAYPGELLAVMGSSGAGKTTLLNALAFRSPQGIQVSPSGMRLLNG
                     QPVDAKEMQARCAYVQQDDLFIGSLTAREHLIFQAMVRMPRHLTYRQRVARVDQVIQEL
                     SLSKCQHTIIGVPGRVKGLSGGERKRLAFASEALTDPPLLICDEPTSGLDSFTAHSVVQ
                     VLKKLSQKGKTVILTIHQPSSELFELFDKILLMAEGRVAFLGTPSEAVDFFSYVGAQCP
                     TNYNPADFYVQVLAVVPGREIESRDRIAKICDNFAISKVARDMEQLLATKNLEKPLEQP
                     ENGYTYKATWFMQFRAVLWRSWLSVLKEPLLVKVRLIQTTMVAILIGLIFLGQQLTQVG
                     VMNINGAIFLFLTNMTFQNVFATINVFTSELPVFMREARSRLYRCDTYFLGKTIAELPL
                     FLTVPLVFTAIAYPMIGLRAGVLHFFNCLALVTLVANVSTSFGYLISCASSSTSMALSV
                     GPPVIIPFLLFGGFFLNSGSVPVYLKWLSYLSWFRYANEGLLINQWADVEPGEISCTSS
                     NTTCPSSGKVILETLNFSAADLPLDYVGLAILIVSFRVLAYLALRLRARRKE"
     protein_bind    4809..4842
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    4875..4969
                     /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        4988..5226
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     primer_bind     5173..5194
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     CDS             join(5250..5252,5253..5255,5256..5969)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5295..5316)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5556..5575)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5903..5924
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             5974..10077
                     /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             5974..10077
                     /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             5974..10077
                     /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             10090..10110
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          10197..10262
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             10392..10412
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    10684..10805
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10721..10740)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10775..10794
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    10865..10934
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     promoter        complement(11153..11171)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(11192..11208)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11192..11208)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11205..11227)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11216..11232
                     /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(11240..11246,11247..11264,11265..11270))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11285..11306
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11423..11440)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11594..12182)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11674..11693)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12353..13144,13145..13213))
                     /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     12976..12995
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13214..13318)
                     /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 cggggttaga 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 gattttccag gccatggtgc
     2221 ggatgccacg acatctgacc tatcggcagc gagtggcccg cgtggatcag gtgatccagg
     2281 agctttcgct cagcaaatgt cagcacacga tcatcggtgt gcccggcagg gtgaaaggtc
     2341 tgtccggcgg agaaaggaag cgtctggcat tcgcctccga ggcactaacc gatccgccgc
     2401 ttctgatctg cgatgagccc acctccggac tggactcatt 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 agcattacat tacatctcaa ctcctatcca gcgtgggtgc ccagtgtcct
     2701 accaactaca atccggcgga cttttacgta caggtgttgg ccgttgtgcc cggacgggag
     2761 atcgagtccc gtgatcggat cgccaagata tgcgacaatt ttgctattag caaagtagcc
     2821 cgggatatgg agcagttgtt ggccaccaaa aatttggaga agccactgga gcagccggag
     2881 aatgggtaca cctacaaggc cacctggttc atgcagttcc gggcggtcct gtggcgatcc
     2941 tggctgtcgg tgctcaagga accactcctc gtaaaagtgc gacttattca gacaacggtg
     3001 agtggttcca gtggaaacaa atgatataac gcttacaatt cttggaaaca aattcgctag
     3061 attttagtta gaattgcctg attccacacc cttcttagtt tttttcaatg agatgtatag
     3121 tttatagttt tgcagaaaat aaataaattt catttaactc gcgaacatgt tgaagatatg
     3181 aatattaatg agatgcgagt aacattttaa tttgcagatg gttgccatct tgattggcct
     3241 catctttttg ggccaacaac tcacgcaagt gggcgtgatg aatatcaacg gagccatctt
     3301 cctcttcctg accaacatga cctttcaaaa cgtctttgcc acgataaatg taagtcttgt
     3361 ttagaataca tttgcatatt aataatttac taactttcta atgaatcgat tcgatttagg
     3421 tgttcacctc agagctgcca gtttttatga gggaggcccg aagtcgactt tatcgctgtg
     3481 acacatactt tctgggcaaa acgattgccg aattaccgct ttttctcaca gtgccactgg
     3541 tcttcacggc gattgcctat ccgatgatcg gactgcgggc cggagtgctg cacttcttca
     3601 actgcctggc gctggtcact ctggtggcca atgtgtcaac gtccttcgga tatctaatat
     3661 cctgcgccag ctcctcgacc tcgatggcgc tgtctgtggg tccgccggtt atcataccat
     3721 tcctgctctt tggcggcttc ttcttgaact cgggctcggt gccagtatac ctcaaatggt
     3781 tgtcgtacct ctcatggttc cgttacgcca acgagggtct gctgattaac caatgggcgg
     3841 acgtggagcc gggcgaaatt agctgcacat cgtcgaacac cacgtgcccc agttcgggca
     3901 aggtcatcct ggagacgctt aacttctccg ccgccgatct gccgctggac tacgtgggtc
     3961 tggccattct catcgtgagc ttccgggtgc tcgcatatct ggctctaaga cttcgggccc
     4021 gacgcaagga gtagccgaca tatatccgaa ataactgctt gttttttttt tttaccatta
     4081 ttaccatcgt gtttactgtt tattgccccc tcaaaaagct aatgtaatta tatttgtgcc
     4141 aataaaaaca agatatgacc tatagaatac aagtatttcc ccttcgaaca tccccacaag
     4201 tagactttgg atttgtcttc taaccaaaag acttacacac ctgcatacct tacatcaaaa
     4261 actcgtttat cgctacataa aacaccggga tatatttttt atatacatac ttttcaaatc
     4321 gcgcgccctc ttcataattc acctccacca caccacgttt cgtagttgct ctttcgctgt
     4381 ctcccacccg ctctccgcaa cacattcacc ttttgttcga cgaccttgga gcgactgtcg
     4441 ttagttccgc gcgattcggt tcgctcaaat ggttccgagt ggttcatttc gtctcaatag
     4501 aaattagtaa taaatatttg tatgtacaat ttatttgctc caatatattt gtatatattt
     4561 ccctcacagc tatatttatt ctaatttaat attatgactt tttaaggtaa ttttttgtga
     4621 cctgttcgga gtgattagcg ttacaatttg aactgaaagt gacatccagt gtttgttcct
     4681 tgtgtagatg catctcaaaa aaatggtggg cataatagtg ttgtttatat atatcaaaaa
     4741 taacaactat aataataaga atacatttaa tttagaaaat gcttggattt cactggaact
     4801 aggctagcat aacttcgtat aatgtatgct atacgaagtt atgctagcgg atccaagctt
     4861 gcatgcctgc aggtcggagt actgtcctcc gagcggagta ctgtcctccg agcggagtac
     4921 tgtcctccga gcggagtact gtcctccgag cggagtactg tcctccgagc ggagactcta
     4981 gcgagcgccg gagtataaat agaggcgctt cgtctacgga gcgacaattc aattcaaaca
     5041 agcaaagtga acacgtcgct aagcgaaagc taagcaaata aacaagcgca gctgaacaag
     5101 ctaaacaatc tgcagtaaag tgcaagttaa agtgaatcaa ttaaaagtaa ccagcaacca
     5161 agtaaatcaa ctgcaactac tgaaatctgc caagaagtaa ttattgaata caagaagaga
     5221 actctgaata gggaattggg aattccaaca tggtgagcaa gggcgaggag ctgttcaccg
     5281 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     5341 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     5401 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
     5461 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     5521 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     5581 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     5641 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     5701 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     5761 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     5821 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccaag ctgagcaaag
     5881 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     5941 ctctcggcat ggacgagctg tacaagtaat aacatggaca agaagtactc cattgggctc
     6001 gatatcggca caaacagcgt cggctgggcc gtcattacgg acgagtacaa ggtgccgagc
     6061 aaaaaattca aagttctggg caataccgat cgccacagca taaagaagaa cctcattggc
     6121 gccctcctgt tcgactccgg ggagacggcc gaagccacgc ggctcaaaag aacagcacgg
     6181 cgcagatata cccgcagaaa gaatcggatc tgctacctgc aggagatctt tagtaatgag
     6241 atggctaagg tggatgactc tttcttccat aggctggagg agtccttttt ggtggaggag
     6301 gataaaaagc acgagcgcca cccaatcttt ggcaatatcg tggacgaggt ggcgtaccat
     6361 gaaaagtacc caaccatata tcatctgagg aagaagcttg tagacagtac tgataaggct
     6421 gacttgcggt tgatctatct cgcgctggcg catatgatca aatttcgggg acacttcctc
     6481 atcgaggggg acctgaaccc agacaacagc gatgtcgaca aactctttat ccaactggtt
     6541 cagacttaca atcagctttt cgaagagaac ccgatcaacg catccggagt tgacgccaaa
     6601 gcaatcctga gcgctaggct gtccaaatcc cggcggctcg aaaacctcat cgcacagctc
     6661 cctggggaga agaagaacgg cctgtttggt aatcttatcg ccctgtcact cgggctgacc
     6721 cccaacttta aatctaactt cgacctggcc gaagatgcca agcttcaact gagcaaagac
     6781 acctacgatg atgatctcga caatctgctg gcccagatcg gcgaccagta cgcagacctt
     6841 tttttggcgg caaagaacct gtcagacgcc attctgctga gtgatattct gcgagtgaac
     6901 acggagatca ccaaagctcc gctgagcgct agtatgatca agcgctatga tgagcaccac
     6961 caagacttga ctttgctgaa ggcccttgtc agacagcaac tgcctgagaa gtacaaggaa
     7021 attttcttcg atcagtctaa aaatggctac gccggataca ttgacggcgg agcaagccag
     7081 gaggaatttt acaaatttat taagcccatc ttggaaaaaa tggacggcac cgaggagctg
     7141 ctggtaaagc ttaacagaga agatctgttg cgcaaacagc gcactttcga caatggaagc
     7201 atcccccacc agattcacct gggcgaactg cacgctatcc tcaggcggca agaggatttc
     7261 tacccctttt tgaaagataa cagggaaaag attgagaaaa tcctcacatt tcggataccc
     7321 tactatgtag gccccctcgc ccggggaaat tccagattcg cgtggatgac tcgcaaatca
     7381 gaagagacca tcactccctg gaacttcgag gaagtcgtgg ataagggggc ctctgcccag
     7441 tccttcatcg aaaggatgac taactttgat aaaaatctgc ctaacgaaaa ggtgcttcct
     7501 aaacactctc tgctgtacga gtacttcaca gtttataacg agctcaccaa ggtcaaatac
     7561 gtcacagaag ggatgagaaa gccagcattc ctgtctggag agcagaagaa agctatcgtg
     7621 gacctcctct tcaagacgaa ccggaaagtt accgtgaaac agctcaaaga agactatttc
     7681 aaaaagattg aatgtttcga ctctgttgaa atcagcggag tggaggatcg cttcaacgca
     7741 tccctgggaa cgtatcacga tctcctgaaa atcattaaag acaaggactt cctggacaat
     7801 gaggagaacg aggacattct tgaggacatt gtcctcaccc ttacgttgtt tgaagatagg
     7861 gagatgattg aagaacgctt gaaaacttac gctcatctct tcgacgacaa agtcatgaaa
     7921 cagctcaaga ggcgccgata tacaggatgg gggcggctgt caagaaaact gatcaatggg
     7981 atccgagaca agcagagtgg aaagacaatc ctggattttc ttaagtccga tggatttgcc
     8041 aaccggaact tcatgcagtt gatccatgat gactctctca cctttaagga ggacatccag
     8101 aaagcacaag tttctggcca gggggacagt cttcacgagc acatcgctaa tcttgcaggt
     8161 agcccagcta tcaaaaaggg aatactgcag accgttaagg tcgtggatga actcgtcaaa
     8221 gtaatgggaa ggcataagcc cgagaatatc gttatcgaga tggcccgaga gaaccaaact
     8281 acccagaagg gacagaagaa cagtagggaa aggatgaaga ggattgaaga gggtataaaa
     8341 gaactggggt cccaaatcct taaggaacac ccagttgaaa acacccagct tcagaatgag
     8401 aagctctacc tgtactacct gcagaacggc agggacatgt acgtggatca ggaactggac
     8461 atcaatcggc tctccgacta cgacgtggat catatcgtgc cccagtcttt tctcaaagat
     8521 gattctattg ataataaagt gttgacaaga tccgataaaa atagagggaa gagtgataac
     8581 gtcccctcag aagaagttgt caagaaaatg aaaaattatt ggcggcagct gctgaacgcc
     8641 aaactgatca cacaacggaa gttcgataat ctgactaagg ctgaacgagg tggcctgtct
     8701 gagttggata aagccggctt catcaaaagg cagcttgttg agacacgcca gatcaccaag
     8761 cacgtggccc aaattctcga ttcacgcatg aacaccaagt acgatgaaaa tgacaaactg
     8821 attcgagagg tgaaagttat tactctgaag tctaagctgg tctcagattt cagaaaggac
     8881 tttcagtttt ataaggtgag agagatcaac aattaccacc atgcgcatga tgcctacctg
     8941 aatgcagtgg taggcactgc acttatcaaa aaatatccca agcttgaatc tgaatttgtt
     9001 tacggagact ataaagtgta cgatgttagg aaaatgatcg caaagtctga gcaggaaata
     9061 ggcaaggcca ccgctaagta cttcttttac agcaatatta tgaatttttt caagaccgag
     9121 attacactgg ccaatggaga gattcggaag cgaccactta tcgaaacaaa cggagaaaca
     9181 ggagaaatcg tgtgggacaa gggtagggat ttcgcgacag tccggaaggt cctgtccatg
     9241 ccgcaggtga acatcgttaa aaagaccgaa gtacagaccg gaggcttctc caaggaaagt
     9301 atcctcccga aaaggaacag cgacaagctg atcgcacgca aaaaagattg ggaccccaag
     9361 aaatacggcg gattcgattc tcctacagtc gcttacagtg tactggttgt ggccaaagtg
     9421 gagaaaggga agtctaaaaa actcaaaagc gtcaaggaac tgctgggcat cacaatcatg
     9481 gagcgatcaa gcttcgaaaa aaaccccatc gactttctcg aggcgaaagg atataaagag
     9541 gtcaaaaaag acctcatcat taagcttccc aagtactctc tctttgagct tgaaaacggc
     9601 cggaaacgaa tgctcgctag tgcgggcgag ctgcagaaag gtaacgagct ggcactgccc
     9661 tctaaatacg ttaatttctt gtatctggcc agccactatg aaaagctcaa agggtctccc
     9721 gaagataatg agcagaagca gctgttcgtg gaacaacaca aacactacct tgatgagatc
     9781 atcgagcaaa taagcgaatt ctccaaaaga gtgatcctcg ccgacgctaa cctcgataag
     9841 gtgctttctg cttacaataa gcacagggat aagcccatca gggagcaggc agaaaacatt
     9901 atccacttgt ttactctgac caacttgggc gcgcctgcag ccttcaagta cttcgacacc
     9961 accatagaca gaaagcggta cacctctaca aaggaggtcc tggacgccac actgattcat
    10021 cagtcaatta cggggctcta tgaaacaaga atcgacctct ctcagctcgg tggagacagc
    10081 agggctgacc ccaagaagaa gaggaaggtg tgagctagcg gatctttgtg aaggaacctt
    10141 acttctgtgg tgtgacataa ttggacaaac tacctacaga gatttaaagc tctaaggtaa
    10201 atataaaatt tttaagtgta taatgtgtta aactactgat tctaattgtt tgtgtatttt
    10261 agattccaac ctatggaact gatgaatggg agcagtggtg gaatgccttt aatgaggaaa
    10321 acctgttttg ctcagaagaa atgccatcta gtgatgatga ggctactgct gactctcaac
    10381 attctactcc tccaaaaaag aagagaaagg tagaagaccc caaggacttt ccttcagaat
    10441 tgctaagttt tttgagtcat gctgtgttta gtaatagaac tcttgcttgc tttgctattt
    10501 acaccacaaa ggaaaaagct gcactgctat acaagaaaat tatggaaaaa tatttgatgt
    10561 atagtgcctt gactagagat cataatcagc cataccacat ttgtagaggt tttacttgct
    10621 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
    10681 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
    10741 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
    10801 tcttatcatg tctctcgagc tcggtaagct tagatccact agtgtcgacg atgtaggtca
    10861 cggtctcgaa gccgcggtgc gggtgccagg gcgtgccctt gggctccccg ggcgcgtact
    10921 ccacctcacc catctggtcc atcatgatga acgggtcgag gtggcggtag ttgatcccgg
    10981 cgaacgcgcg gcgcaccggg aagccctcgc cctcgaaacc gctgggcgcg gtggtcacgg
    11041 tgagcacggg acgtgcgacg gcgtcggcgg gtgcggatac gcggggcagc gtcagcgggt
    11101 tctcgacggt cacggcgggc atgtcgacac tagttctagc cagcttttgt tccctttagt
    11161 gagggttaat ttcgagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
    11221 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
    11281 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
    11341 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
    11401 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
    11461 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
    11521 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
    11581 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
    11641 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
    11701 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
    11761 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
    11821 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
    11881 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
    11941 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
    12001 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
    12061 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
    12121 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
    12181 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
    12241 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
    12301 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
    12361 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
    12421 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
    12481 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
    12541 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
    12601 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
    12661 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
    12721 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
    12781 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
    12841 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
    12901 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
    12961 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
    13021 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
    13081 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
    13141 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
    13201 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
    13261 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
    13321 catttccccg aaaagtgcca c
//
