LOCUS       sequence-407311-       15370 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of dCBEevoCDA1 under control of Gal4/UAS and 
            FLP/FRT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15370)
  AUTHORS   Doll RM, Boutros M, Port F
  TITLE     A temperature-tolerant CRISPR base editor mediates highly efficient 
            and precise gene editing in Drosophila.
  JOURNAL   Sci Adv. 2023 Sep;9(35):eadj1568. doi: 10.1126/sciadv.adj1568. Epub 
            2023 Aug 30.
  PUBMED    37647411
REFERENCE   2  (bases 1 to 15370)
  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: "Sci Adv."; 
            date: "2023-09"; volume: "9(35):eadj1568. doi"; pages: " 
            10.1126/sciadv.adj1568. Epub 2023 Aug 30"
COMMENT     Sequence Label: pUAS-FRT-GFP-FRT-uM-dCBEevoCDA1
FEATURES             Location/Qualifiers
     source          1..15370
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(540..1331,1332..1400))
                     /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     1163..1182
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1401..1505)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1532..1986)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1668..1689)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1880..1899
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2112..2134
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2126..2143
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2127..2143
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2150..2169
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2150..2168
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     gene            2193..6329
                     /gene="white"
                     /label=mini-white
                     /note="This modified version of the white gene lacks part 
                     of the first intron."
     primer_bind     complement(2202..2220)
                     /label=pCasper-F
                     /note="Drosophila mini-white gene, reverse primer"
     CDS             join(2672..2743,3145..3418,3493..4147,4209..4524,4745..4876,
                     4947..5561)
                     /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    6336..6369
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    6402..6496
                     /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        6515..6753
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     primer_bind     6700..6721
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     protein_bind    complement(6787..6820)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             join(6825..6827,6828..6830,6831..7541)
                     /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(7131..7150)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7478..7499
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     intron          7649..7714
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             7844..7864
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    8136..8270
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(8288..8321)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             8502..8522
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(9243..9266,9267..9269,9270..13343)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             9243..13343
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             13374..13622
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             13665..13685
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          13772..13837
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             13967..13987
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    14259..14393
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    14422..14491
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     promoter        complement(14710..14728)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(14749..14765)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(14749..14765)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(14762..14784)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    14773..14789
                     /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(14797..14803,14804..14821,14822..14827))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    14842..14863
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(14980..14997)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(15151..15370,1..369))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(15231..15250)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
       61 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
      121 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
      181 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
      241 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
      301 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
      361 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
      421 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
      481 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
      541 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
      601 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
      661 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
      721 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
      781 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
      841 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
      901 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
      961 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     1021 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     1081 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     1141 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     1201 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     1261 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     1321 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     1381 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     1441 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     1501 ccgcgcacat ttccccgaaa agtgccacct gaattgtaag cgttaatatt ttgttaaaat
     1561 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     1621 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     1681 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     1741 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     1801 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     1861 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     1921 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     1981 gcgcgtccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     2041 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     2101 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgaattgt aatacgactc
     2161 actatagggc gaattgggta cgtaccgggc ccctagtatg tatgtaagtt aataaaaccc
     2221 atttttgcgg aaagtagata aaaaaaacat tttttttttt tactgcactg gatatcattg
     2281 aacttatctg atcagtttta aatttacttc gatccaaggg tatttgatgt accaggttct
     2341 ttcgattacc tctcactcaa aatgacattc cactcaaagt cagcgctgtt tgcctccttc
     2401 tctgtccaca gaaatatcgc cgtctctttc gccgctgcgt ccgctatctc tttcgccacc
     2461 gtttgtagcg ttacgtagcg tcaatgtccg ccttcagttg cattttgtca gcggtttcgt
     2521 gacgaagctc caagcggttt acgccatcaa ttaaacacaa agtgctgtgc caaaactcct
     2581 ctcgcttctt atttttgttt gttttttgag tgattggggt ggtgattggt tttgggtggg
     2641 taagcagggg aaagtgtgaa aaatcccggc aatgggccaa gaggatcagg agctattaat
     2701 tcgcggaggc agcaaacacc catctgccga gcatctgaac aatgtgagta gtacatgtgc
     2761 atacatctta agttcacttg atctatagga actgcgattg caacatcaaa ttgtctgcgg
     2821 cgtgagaact gcgacccaca aaaatcccaa accgcaattg cacaaacaaa tagtgacacg
     2881 aaacagatta ttctggtagc tgttctcgct atataagaca atttttgaga tcatatcatg
     2941 atcaagacat ctaaaggcat tcattttcga ctatattctt ttttacaaaa aatataacaa
     3001 ccagatattt taagctgatc ctagatgcac aaaaaataaa taaaagtata aacctacttc
     3061 gtaggatact tcggggtact ttttgttcgg ggttagatga gcataacgct tgtagttgat
     3121 atttgagatc ccctatcatt gcagggtgac agcggagcgg cttcgcagag ctgcattaac
     3181 cagggcttcg ggcaggccaa aaactacggc acgctccggc cacccagtcc gccggaggac
     3241 tccggttcag ggagcggcca actagccgag aacctcacct atgcctggca caatatggac
     3301 atctttgggg cggtcaatca gccgggctcc ggatggcggc agctggtcaa ccggacacgc
     3361 ggactattct gcaacgagcg acacataccg gcgcccagga aacatttgct caagaacggt
     3421 gagtttctat tcgcagtcgg ctgatctgtg tgaaatctta ataaagggtc caattaccaa
     3481 tttgaaactc agtttgcggc gtggcctatc cgggcgaact tttggccgtg atgggcagtt
     3541 ccggtgccgg aaagacgacc ctgctgaatg cccttgcctt tcgatcgccg cagggcatcc
     3601 aagtatcgcc atccgggatg cgactgctca atggccaacc tgtggacgcc aaggagatgc
     3661 aggccaggtg cgcctatgtc cagcaggatg acctctttat cggctcccta acggccaggg
     3721 aacacctgat tttccaggcc atggtgcgga tgccacgaca tctgacctat cggcagcgag
     3781 tggcccgcgt ggatcaggtg atccaggagc tttcgctcag caaatgtcag cacacgatca
     3841 tcggtgtgcc cggcagggtg aaaggtctgt ccggcggaga aaggaagcgt ctggcattcg
     3901 cctccgaggc actaaccgat ccgccgcttc tgatctgcga tgagcccacc tccggactgg
     3961 actcatttac cgcccacagc gtcgtccagg tgctgaagaa gctgtcgcag aagggcaaga
     4021 ccgtcatcct gaccattcat cagccgtctt ccgagctgtt tgagctcttt gacaagatcc
     4081 ttctgatggc cgagggcagg gtagctttct tgggcactcc cagcgaagcc gtcgacttct
     4141 tttcctagtg agttcgatgt gtttattaag ggtatctagc attacattac atctcaactc
     4201 ctatccagcg tgggtgccca gtgtcctacc aactacaatc cggcggactt ttacgtacag
     4261 gtgttggccg ttgtgcccgg acgggagatc gagtcccgtg atcggatcgc caagatatgc
     4321 gacaattttg ctattagcaa agtagcccgg gatatggagc agttgttggc caccaaaaat
     4381 ttggagaagc cactggagca gccggagaat gggtacacct acaaggccac ctggttcatg
     4441 cagttccggg cggtcctgtg gcgatcctgg ctgtcggtgc tcaaggaacc actcctcgta
     4501 aaagtgcgac ttattcagac aacggtgagt ggttccagtg gaaacaaatg atataacgct
     4561 tacaattctt ggaaacaaat tcgctagatt ttagttagaa ttgcctgatt ccacaccctt
     4621 cttagttttt ttcaatgaga tgtatagttt atagttttgc agaaaataaa taaatttcat
     4681 ttaactcgcg aacatgttga agatatgaat attaatgaga tgcgagtaac attttaattt
     4741 gcagatggtt gccatcttga ttggcctcat ctttttgggc caacaactca cgcaagtggg
     4801 cgtgatgaat atcaacggag ccatcttcct cttcctgacc aacatgacct ttcaaaacgt
     4861 ctttgccacg ataaatgtaa gtcttgttta gaatacattt gcatattaat aatttactaa
     4921 ctttctaatg aatcgattcg atttaggtgt tcacctcaga gctgccagtt tttatgaggg
     4981 aggcccgaag tcgactttat cgctgtgaca catactttct gggcaaaacg attgccgaat
     5041 taccgctttt tctcacagtg ccactggtct tcacggcgat tgcctatccg atgatcggac
     5101 tgcgggccgg agtgctgcac ttcttcaact gcctggcgct ggtcactctg gtggccaatg
     5161 tgtcaacgtc cttcggatat ctaatatcct gcgccagctc ctcgacctcg atggcgctgt
     5221 ctgtgggtcc gccggttatc ataccattcc tgctctttgg cggcttcttc ttgaactcgg
     5281 gctcggtgcc agtatacctc aaatggttgt cgtacctctc atggttccgt tacgccaacg
     5341 agggtctgct gattaaccaa tgggcggacg tggagccggg cgaaattagc tgcacatcgt
     5401 cgaacaccac gtgccccagt tcgggcaagg tcatcctgga gacgcttaac ttctccgccg
     5461 ccgatctgcc gctggactac gtgggtctgg ccattctcat cgtgagcttc cgggtgctcg
     5521 catatctggc tctaagactt cgggcccgac gcaaggagta gccgacatat atccgaaata
     5581 actgcttgtt tttttttttt accattatta ccatcgtgtt tactgtttat tgccccctca
     5641 aaaagctaat gtaattatat ttgtgccaat aaaaacaaga tatgacctat agaatacaag
     5701 tatttcccct tcgaacatcc ccacaagtag actttggatt tgtcttctaa ccaaaagact
     5761 tacacacctg cataccttac atcaaaaact cgtttatcgc tacataaaac accgggatat
     5821 attttttata tacatacttt tcaaatcgcg cgccctcttc ataattcacc tccaccacac
     5881 cacgtttcgt agttgctctt tcgctgtctc ccacccgctc tccgcaacac attcaccttt
     5941 tgttcgacga ccttggagcg actgtcgtta gttccgcgcg attcggttcg ctcaaatggt
     6001 tccgagtggt tcatttcgtc tcaatagaaa ttagtaataa atatttgtat gtacaattta
     6061 tttgctccaa tatatttgta tatatttccc tcacagctat atttattcta atttaatatt
     6121 atgacttttt aaggtaattt tttgtgacct gttcggagtg attagcgtta caatttgaac
     6181 tgaaagtgac atccagtgtt tgttccttgt gtagatgcat ctcaaaaaaa tggtgggcat
     6241 aatagtgttg tttatatata tcaaaaataa caactataat aataagaata catttaattt
     6301 agaaaatgct tggatttcac tggaactagg ctagcataac ttcgtataat gtatgctata
     6361 cgaagttatg ctagcggatc caagcttgca tgcctgcagg tcggagtact gtcctccgag
     6421 cggagtactg tcctccgagc ggagtactgt cctccgagcg gagtactgtc ctccgagcgg
     6481 agtactgtcc tccgagcgga gactctagcg agcgccggag tataaataga ggcgcttcgt
     6541 ctacggagcg acaattcaat tcaaacaagc aaagtgaaca cgtcgctaag cgaaagctaa
     6601 gcaaataaac aagcgcagct gaacaagcta aacaatctgc agtaaagtgc aagttaaagt
     6661 gaatcaatta aaagtaacca gcaaccaagt aaatcaactg caactactga aatctgccaa
     6721 gaagtaatta ttgaatacaa gaagagaact ctgaataggg aattgggaag gtaccgcata
     6781 ccaaacgaag ttcctatact ttctagagaa taggaacttc caaaatggtg agcaagggcg
     6841 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     6901 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     6961 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     7021 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     7081 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     7141 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     7201 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     7261 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     7321 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     7381 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     7441 ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     7501 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gccgcggccc gagaagcgta
     7561 aacaggaata aggatctttg tgaaggaacc ttacttctgt ggtgtgacat aattggacaa
     7621 actacctaca gagatttaaa gctctaaggt aaatataaaa tttttaagtg tataatgtgt
     7681 taaactactg attctaattg tttgtgtatt ttagattcca acctatggaa ctgatgaatg
     7741 ggagcagtgg tggaatgcct ttaatgagga aaacctgttt tgctcagaag aaatgccatc
     7801 tagtgatgat gaggctactg ctgactctca acattctact cctccaaaaa agaagagaaa
     7861 ggtagaagac cccaaggact ttccttcaga attgctaagt tttttgagtc atgctgtgtt
     7921 tagtaataga actcttgctt gctttgctat ttacaccaca aaggaaaaag ctgcactgct
     7981 atacaagaaa attatggaaa aatatttgat gtatagtgcc ttgactagag atcataatca
     8041 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     8101 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     8161 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     8221 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc actagtgatc
     8281 tggccgggaa gttcctatac tttctagaga ataggaactt cgaattccaa catggtgagc
     8341 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggctctaga
     8401 gccctcgccg gacacgctga acttgtggcc gtttacgtcg ctagataata aggtaccttc
     8461 ccaccatgaa acggacagcc gacggaagcg agttcgagtc accaaagaag aagcggaaag
     8521 tcagtaccga cgccgagtac gtgcggatcc acgagaagct ggatatctat acattcaaga
     8581 agcagtttag caacaataag aagtccgtgt ctcacagatg ctacgtgctg ttcgagctga
     8641 agcggagagg agagaggcgc gcctgttttt ggggctatgc cgtgaacaag ccacagtctg
     8701 gaaccgagag gggaatccac gcagagatct tcagcatcag gaaggtggag gagtacctgc
     8761 gcgacaaccc cggccagttt acaatcaatt ggtatagctc ctggagccct tgcgccgatt
     8821 gtgccgagaa gatcctggag tggtacaacc aggagctgag gggcaatggc cacaccctga
     8881 agatctgggt gtgcaagctg tactatgaga agaacgccag gaatcagatc ggcctgtgga
     8941 acctgcgcga caatggcgtg ggcctgaacg tgatggtgtc cgagcactat cagtgctgtc
     9001 gcaagatctt tatccagtct agccacaatc agctgaacga gaatcggtgg ctggagaaga
     9061 cactgaagag agccgagaag cggagaagcg agctgtccat catgtttcag gtgaagatcc
     9121 tgcacaccac aaagtctccc gccgtgtctg gcggatctag cggaggatcc tctggcagcg
     9181 agacaccagg aacaagcgag tcagcaacac cagagagcag tggcggcagc agcggcggca
     9241 gcgacaagaa gtactccatt gggctcgcca tcggcacaaa cagcgtcggc tgggccgtca
     9301 ttacggacga gtacaaggtg ccgagcaaaa aattcaaagt tctgggcaat accgatcgcc
     9361 acagcataaa gaagaacctc attggcgccc tcctgttcga ctccggggag acggccgaag
     9421 ccacgcggct caaaagaaca gcacggcgca gatatacccg cagaaagaat cggatctgct
     9481 acctgcagga gatctttagt aatgagatgg ctaaggtgga tgactctttc ttccataggc
     9541 tggaggagtc ctttttggtg gaggaggata aaaagcacga gcgccaccca atctttggca
     9601 atatcgtgga cgaggtggcg taccatgaaa agtacccaac catatatcat ctgaggaaga
     9661 agcttgtaga cagtactgat aaggctgact tgcggttgat ctatctcgcg ctggcgcata
     9721 tgatcaaatt tcggggacac ttcctcatcg agggggacct gaacccagac aacagcgatg
     9781 tcgacaaact ctttatccaa ctggttcaga cttacaatca gcttttcgaa gagaacccga
     9841 tcaacgcatc cggagttgac gccaaagcaa tcctgagcgc taggctgtcc aaatcccggc
     9901 ggctcgaaaa cctcatcgca cagctccctg gggagaagaa gaacggcctg tttggtaatc
     9961 ttatcgccct gtcactcggg ctgaccccca actttaaatc taacttcgac ctggccgaag
    10021 atgccaagct tcaactgagc aaagacacct acgatgatga tctcgacaat ctgctggccc
    10081 agatcggcga ccagtacgca gacctttttt tggcggcaaa gaacctgtca gacgccattc
    10141 tgctgagtga tattctgcga gtgaacacgg agatcaccaa agctccgctg agcgctagta
    10201 tgatcaagcg ctatgatgag caccaccaag acttgacttt gctgaaggcc cttgtcagac
    10261 agcaactgcc tgagaagtac aaggaaattt tcttcgatca gtctaaaaat ggctacgccg
    10321 gatacattga cggcggagca agccaggagg aattttacaa atttattaag cccatcttgg
    10381 aaaaaatgga cggcaccgag gagctgctgg taaagcttaa cagagaagat ctgttgcgca
    10441 aacagcgcac tttcgacaat ggaagcatcc cccaccagat tcacctgggc gaactgcacg
    10501 ctatcctcag gcggcaagag gatttctacc cctttttgaa agataacagg gaaaagattg
    10561 agaaaatcct cacatttcgg ataccctact atgtaggccc cctcgcccgg ggaaattcca
    10621 gattcgcgtg gatgactcgc aaatcagaag agaccatcac tccctggaac ttcgaggaag
    10681 tcgtggataa gggggcctct gcccagtcct tcatcgaaag gatgactaac tttgataaaa
    10741 atctgcctaa cgaaaaggtg cttcctaaac actctctgct gtacgagtac ttcacagttt
    10801 ataacgagct caccaaggtc aaatacgtca cagaagggat gagaaagcca gcattcctgt
    10861 ctggagagca gaagaaagct atcgtggacc tcctcttcaa gacgaaccgg aaagttaccg
    10921 tgaaacagct caaagaagac tatttcaaaa agattgaatg tttcgactct gttgaaatca
    10981 gcggagtgga ggatcgcttc aacgcatccc tgggaacgta tcacgatctc ctgaaaatca
    11041 ttaaagacaa ggacttcctg gacaatgagg agaacgagga cattcttgag gacattgtcc
    11101 tcacccttac gttgtttgaa gatagggaga tgattgaaga acgcttgaaa acttacgctc
    11161 atctcttcga cgacaaagtc atgaaacagc tcaagaggcg ccgatataca ggatgggggc
    11221 ggctgtcaag aaaactgatc aatgggatcc gagacaagca gagtggaaag acaatcctgg
    11281 attttcttaa gtccgatgga tttgccaacc ggaacttcat gcagttgatc catgatgact
    11341 ctctcacctt taaggaggac atccagaaag cacaagtttc tggccagggg gacagtcttc
    11401 acgagcacat cgctaatctt gcaggtagcc cagctatcaa aaagggaata ctgcagaccg
    11461 ttaaggtcgt ggatgaactc gtcaaagtaa tgggaaggca taagcccgag aatatcgtta
    11521 tcgagatggc ccgagagaac caaactaccc agaagggaca gaagaacagt agggaaagga
    11581 tgaagaggat tgaagagggt ataaaagaac tggggtccca aatccttaag gaacacccag
    11641 ttgaaaacac ccagcttcag aatgagaagc tctacctgta ctacctgcag aacggcaggg
    11701 acatgtacgt ggatcaggaa ctggacatca atcggctctc cgactacgac gtggatcata
    11761 tcgtgcccca gtcttttctc aaagatgatt ctattgataa taaagtgttg acaagatccg
    11821 ataaaaatag agggaagagt gataacgtcc cctcagaaga agttgtcaag aaaatgaaaa
    11881 attattggcg gcagctgctg aacgccaaac tgatcacaca acggaagttc gataatctga
    11941 ctaaggctga acgaggtggc ctgtctgagt tggataaagc cggcttcatc aaaaggcagc
    12001 ttgttgagac acgccagatc accaagcacg tggcccaaat tctcgattca cgcatgaaca
    12061 ccaagtacga tgaaaatgac aaactgattc gagaggtgaa agttattact ctgaagtcta
    12121 agctggtctc agatttcaga aaggactttc agttttataa ggtgagagag atcaacaatt
    12181 accaccatgc gcatgatgcc tacctgaatg cagtggtagg cactgcactt atcaaaaaat
    12241 atcccaagct tgaatctgaa tttgtttacg gagactataa agtgtacgat gttaggaaaa
    12301 tgatcgcaaa gtctgagcag gaaataggca aggccaccgc taagtacttc ttttacagca
    12361 atattatgaa ttttttcaag accgagatta cactggccaa tggagagatt cggaagcgac
    12421 cacttatcga aacaaacgga gaaacaggag aaatcgtgtg ggacaagggt agggatttcg
    12481 cgacagtccg gaaggtcctg tccatgccgc aggtgaacat cgttaaaaag accgaagtac
    12541 agaccggagg cttctccaag gaaagtatcc tcccgaaaag gaacagcgac aagctgatcg
    12601 cacgcaaaaa agattgggac cccaagaaat acggcggatt cgattctcct acagtcgctt
    12661 acagtgtact ggttgtggcc aaagtggaga aagggaagtc taaaaaactc aaaagcgtca
    12721 aggaactgct gggcatcaca atcatggagc gatcaagctt cgaaaaaaac cccatcgact
    12781 ttctcgaggc gaaaggatat aaagaggtca aaaaagacct catcattaag cttcccaagt
    12841 actctctctt tgagcttgaa aacggccgga aacgaatgct cgctagtgcg ggcgagctgc
    12901 agaaaggtaa cgagctggca ctgccctcta aatacgttaa tttcttgtat ctggccagcc
    12961 actatgaaaa gctcaaaggg tctcccgaag ataatgagca gaagcagctg ttcgtggaac
    13021 aacacaaaca ctaccttgat gagatcatcg agcaaataag cgaattctcc aaaagagtga
    13081 tcctcgccga cgctaacctc gataaggtgc tttctgctta caataagcac agggataagc
    13141 ccatcaggga gcaggcagaa aacattatcc acttgtttac tctgaccaac ttgggcgcgc
    13201 ctgcagcctt caagtacttc gacaccacca tagacagaaa gcggtacacc tctacaaagg
    13261 aggtcctgga cgccacactg attcatcagt caattacggg gctctatgaa acaagaatcg
    13321 acctctctca gctcggtgga gacagcggcg gttccggtgg atccggcgga agcacgaacc
    13381 tgagtgatat aattgaaaag gaaaccggta aacagctcgt tatccaggag tcgatcctca
    13441 tgctgcccga ggaagtcgag gaagtcatcg gtaacaaacc ggaatcggat attctggtac
    13501 atacggccta tgatgaatcc accgatgaga atgtaatgtt gttgacatcg gatgcaccgg
    13561 agtataagcc ttgggcgctc gtcatccagg actccaatgg cgagaacaag attaagatgc
    13621 tctccggagg atccaaacga acagctgacg gtagtgaatt cgaaccaaaa aaaaagcgca
    13681 aggtataatc tagaggatct ttgtgaagga accttacttc tgtggtgtga cataattgga
    13741 caaactacct acagagattt aaagctctaa ggtaaatata aaatttttaa gtgtataatg
    13801 tgttaaacta ctgattctaa ttgtttgtgt attttagatt ccaacctatg gaactgatga
    13861 atgggagcag tggtggaatg cctttaatga ggaaaacctg ttttgctcag aagaaatgcc
    13921 atctagtgat gatgaggcta ctgctgactc tcaacattct actcctccaa aaaagaagag
    13981 aaaggtagaa gaccccaagg actttccttc agaattgcta agttttttga gtcatgctgt
    14041 gtttagtaat agaactcttg cttgctttgc tatttacacc acaaaggaaa aagctgcact
    14101 gctatacaag aaaattatgg aaaaatattt gatgtatagt gccttgacta gagatcataa
    14161 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
    14221 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
    14281 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
    14341 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atccactagt
    14401 gtcgacgatg taggtcacgg tctcgaagcc gcggtgcggg tgccagggcg tgcccttggg
    14461 ctccccgggc gcgtactcca cctcacccat ctggtccatc atgatgaacg ggtcgaggtg
    14521 gcggtagttg atcccggcga acgcgcggcg caccgggaag ccctcgccct cgaaaccgct
    14581 gggcgcggtg gtcacggtga gcacgggacg tgcgacggcg tcggcgggtg cggatacgcg
    14641 gggcagcgtc agcgggttct cgacggtcac ggcgggcatg tcgacactag ttctagccag
    14701 cttttgttcc ctttagtgag ggttaatttc gagcttggcg taatcatggt catagctgtt
    14761 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
    14821 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
    14881 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
    14941 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
    15001 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
    15061 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
    15121 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
    15181 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
    15241 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
    15301 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
    15361 gtatctcagt
//
