LOCUS       Exported               15371 bp ds-DNA     circular SYN 03-JUL-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pFP1138 pUASTattB-STOPuORF_evoCDA1_Cas9D10A_1xUGI_KpnI
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 15371)
  AUTHORS   Fillip Port
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jul 3, 2023 from SnapGene 3.3.4
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..15371
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     rep_origin      4..459
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      623..642
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     promoter        623..641
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    667..4802
                     /label=white dominant eye marker
                     /note="white dominant eye marker"
     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 ia 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"
     misc_feature    4809..4842
                     /label=loxP
                     /note="loxP"
     protein_bind    complement(4809..4842)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    4849..5230
                     /label=5xUAS-hsp70
                     /note="5xUAS-hsp70"
     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"
     protein_bind    complement(5260..5293)
                     /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)"
     misc_feature    5260..5293
                     /note="FRT site"
     misc_feature    5294..5297
                     /note="Kozak sequence"
     CDS             5298..6014
                     /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"
     misc_feature    5298..6014
                     /note="mEGFP (A206K)"
     misc_feature    6015..6041
                     /label=NLS
     misc_feature    6021..6041
                     /note="nls UDE1"
     misc_feature    6045..6743
                     /note="SV40 terminator"
     intron          6122..6187
                     /label=small t intron
                     /note="simian virus 40 (SV40) small t antigen intron"
     CDS             6317..6337
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T 
                     antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    6609..6743
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(6761..6794)
                     /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)"
     misc_feature    6761..6794
                     /note="FRT site"
     misc_feature    6795
                     /note="hCas9 (Church Lab)"
     misc_feature    6796..6800
                     /label=MCS
     CDS             6805..6921
                     /codon_start=1
                     /label=uORF
                     /translation="MVSKGEELFTGVVPILVELDGSRALAGHAELVAVYVAR"
     misc_feature    6934..6938
                     /label=Kozak
     misc_signal     6934..6938
                     /label=MCS [Split]
     misc_feature    6934..6938
                     /label=gBlock1
     misc_feature    6939..8211
                     /label=evoCDA1 gBlock
     CDS             6939..6974
                     /codon_start=1
                     /label=Feature 33
                     /translation="MKRTADGSEFES"
     CDS             6975..6995
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    6996..7619
                     /label=evoCDA1
     CDS             6996..7619
                     /codon_start=1
                     /label=evoCDA1
                     /translation="STDAEYVRIHEKLDIYTFKKQFSNNKKSVSHRCYVLFELKRRGER
                     RACFWGYAVNKPQSGTERGIHAEIFSIRKVEEYLRDNPGQFTINWYSSWSPCADCAEKI
                     LEWYNQELRGNGHTLKIWVCKLYYEKNARNQIGLWNLRDNGVGLNVMVSEHYQCCRKIF
                     IQSSHNQLNENRWLEKTLKRAEKRRSELSIMFQVKILHTTKSPAV"
     CDS             7620..7715
                     /codon_start=1
                     /label=32aa linker
                     /translation="SGGSSGGSSGSETPGTSESATPESSGGSSGGS"
     CDS             7716..11816
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /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             11847..12095
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             12138..12158
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    12168..12867
                     /label=SV40 poly(A)
                     /note="SV40 poly(A)"
     intron          12245..12310
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             12440..12460
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    12732..12866
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    12874..13158
                     /label=attB site
                     /note="attB site"
     protein_bind    12895..12964
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     regulatory      13182..13201
                     /regulatory_class="promoter"
                     /note="T3 promoter"
     promoter        complement(13183..13201)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(13222..13238)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    13246..13262
                     /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(13270..13300)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13315..13336
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(13624..14212)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(14383..15177,15208..15243))
                     /codon_start=1
                     /product="ampicillin resistance protein"
                     /label=ampicillin resistance protein
                     /protein_id="ABN09740.1"
                     /translation="MSIQHFRVALIPAHPETLVKVKDAEDQLGARVGYIELDLNSGKIL
                     ESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDG
                     MTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPN
                     DERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADK
                     SGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     promoter        complement(15244..15348)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
       61 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
      121 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
      181 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
      241 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
      301 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
      361 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
      421 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc 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 aaggtaccgc ataccaaacg aagttcctat actttctaga
     5281 gaataggaac ttccaaaatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     5341 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     5401 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     5461 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
     5521 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     5581 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     5641 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     5701 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
     5761 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
     5821 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     5881 tgctgcccga caaccactac ctgagcaccc agtccaagct gagcaaagac cccaacgaga
     5941 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     6001 acgagctgta caagccgcgg cccgagaagc gtaaacagga ataaggatct ttgtgaagga
     6061 accttacttc tgtggtgtga cataattgga caaactacct acagagattt aaagctctaa
     6121 ggtaaatata aaatttttaa gtgtataatg tgttaaacta ctgattctaa ttgtttgtgt
     6181 attttagatt ccaacctatg gaactgatga atgggagcag tggtggaatg cctttaatga
     6241 ggaaaacctg ttttgctcag aagaaatgcc atctagtgat gatgaggcta ctgctgactc
     6301 tcaacattct actcctccaa aaaagaagag aaaggtagaa gaccccaagg actttccttc
     6361 agaattgcta agttttttga gtcatgctgt gtttagtaat agaactcttg cttgctttgc
     6421 tatttacacc acaaaggaaa aagctgcact gctatacaag aaaattatgg aaaaatattt
     6481 gatgtatagt gccttgacta gagatcataa tcagccatac cacatttgta gaggttttac
     6541 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     6601 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     6661 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     6721 atgtatctta tcatgtctgg atcactagtg atctggccgg gaagttccta tactttctag
     6781 agaataggaa cttcgaattc caacatggtg agcaagggcg aggagctgtt caccggggtg
     6841 gtgcccatcc tggtcgagct ggacggctct agagccctcg ccggacacgc tgaacttgtg
     6901 gccgtttacg tcgctagata ataaggtacc ttcccaccat gaaacggaca gccgacggaa
     6961 gcgagttcga gtcaccaaag aagaagcgga aagtcagtac cgacgccgag tacgtgcgga
     7021 tccacgagaa gctggatatc tatacattca agaagcagtt tagcaacaat aagaagtccg
     7081 tgtctcacag atgctacgtg ctgttcgagc tgaagcggag aggagagagg cgcgcctgtt
     7141 tttggggcta tgccgtgaac aagccacagt ctggaaccga gaggggaatc cacgcagaga
     7201 tcttcagcat caggaaggtg gaggagtacc tgcgcgacaa ccccggccag tttacaatca
     7261 attggtatag ctcctggagc ccttgcgccg attgtgccga gaagatcctg gagtggtaca
     7321 accaggagct gaggggcaat ggccacaccc tgaagatctg ggtgtgcaag ctgtactatg
     7381 agaagaacgc caggaatcag atcggcctgt ggaacctgcg cgacaatggc gtgggcctga
     7441 acgtgatggt gtccgagcac tatcagtgct gtcgcaagat ctttatccag tctagccaca
     7501 atcagctgaa cgagaatcgg tggctggaga agacactgaa gagagccgag aagcggagaa
     7561 gcgagctgtc catcatgttt caggtgaaga tcctgcacac cacaaagtct cccgccgtgt
     7621 ctggcggatc tagcggagga tcctctggca gcgagacacc aggaacaagc gagtcagcaa
     7681 caccagagag cagtggcggc agcagcggcg gcagcgacaa gaagtactcc attgggctcg
     7741 ccatcggcac aaacagcgtc ggctgggccg tcattacgga cgagtacaag gtgccgagca
     7801 aaaaattcaa agttctgggc aataccgatc gccacagcat aaagaagaac ctcattggcg
     7861 ccctcctgtt cgactccggg gagacggccg aagccacgcg gctcaaaaga acagcacggc
     7921 gcagatatac ccgcagaaag aatcggatct gctacctgca ggagatcttt agtaatgaga
     7981 tggctaaggt ggatgactct ttcttccata ggctggagga gtcctttttg gtggaggagg
     8041 ataaaaagca cgagcgccac ccaatctttg gcaatatcgt ggacgaggtg gcgtaccatg
     8101 aaaagtaccc aaccatatat catctgagga agaagcttgt agacagtact gataaggctg
     8161 acttgcggtt gatctatctc gcgctggcgc atatgatcaa atttcgggga cacttcctca
     8221 tcgaggggga cctgaaccca gacaacagcg atgtcgacaa actctttatc caactggttc
     8281 agacttacaa tcagcttttc gaagagaacc cgatcaacgc atccggagtt gacgccaaag
     8341 caatcctgag cgctaggctg tccaaatccc ggcggctcga aaacctcatc gcacagctcc
     8401 ctggggagaa gaagaacggc ctgtttggta atcttatcgc cctgtcactc gggctgaccc
     8461 ccaactttaa atctaacttc gacctggccg aagatgccaa gcttcaactg agcaaagaca
     8521 cctacgatga tgatctcgac aatctgctgg cccagatcgg cgaccagtac gcagaccttt
     8581 ttttggcggc aaagaacctg tcagacgcca ttctgctgag tgatattctg cgagtgaaca
     8641 cggagatcac caaagctccg ctgagcgcta gtatgatcaa gcgctatgat gagcaccacc
     8701 aagacttgac tttgctgaag gcccttgtca gacagcaact gcctgagaag tacaaggaaa
     8761 ttttcttcga tcagtctaaa aatggctacg ccggatacat tgacggcgga gcaagccagg
     8821 aggaatttta caaatttatt aagcccatct tggaaaaaat ggacggcacc gaggagctgc
     8881 tggtaaagct taacagagaa gatctgttgc gcaaacagcg cactttcgac aatggaagca
     8941 tcccccacca gattcacctg ggcgaactgc acgctatcct caggcggcaa gaggatttct
     9001 accccttttt gaaagataac agggaaaaga ttgagaaaat cctcacattt cggataccct
     9061 actatgtagg ccccctcgcc cggggaaatt ccagattcgc gtggatgact cgcaaatcag
     9121 aagagaccat cactccctgg aacttcgagg aagtcgtgga taagggggcc tctgcccagt
     9181 ccttcatcga aaggatgact aactttgata aaaatctgcc taacgaaaag gtgcttccta
     9241 aacactctct gctgtacgag tacttcacag tttataacga gctcaccaag gtcaaatacg
     9301 tcacagaagg gatgagaaag ccagcattcc tgtctggaga gcagaagaaa gctatcgtgg
     9361 acctcctctt caagacgaac cggaaagtta ccgtgaaaca gctcaaagaa gactatttca
     9421 aaaagattga atgtttcgac tctgttgaaa tcagcggagt ggaggatcgc ttcaacgcat
     9481 ccctgggaac gtatcacgat ctcctgaaaa tcattaaaga caaggacttc ctggacaatg
     9541 aggagaacga ggacattctt gaggacattg tcctcaccct tacgttgttt gaagataggg
     9601 agatgattga agaacgcttg aaaacttacg ctcatctctt cgacgacaaa gtcatgaaac
     9661 agctcaagag gcgccgatat acaggatggg ggcggctgtc aagaaaactg atcaatggga
     9721 tccgagacaa gcagagtgga aagacaatcc tggattttct taagtccgat ggatttgcca
     9781 accggaactt catgcagttg atccatgatg actctctcac ctttaaggag gacatccaga
     9841 aagcacaagt ttctggccag ggggacagtc ttcacgagca catcgctaat cttgcaggta
     9901 gcccagctat caaaaaggga atactgcaga ccgttaaggt cgtggatgaa ctcgtcaaag
     9961 taatgggaag gcataagccc gagaatatcg ttatcgagat ggcccgagag aaccaaacta
    10021 cccagaaggg acagaagaac agtagggaaa ggatgaagag gattgaagag ggtataaaag
    10081 aactggggtc ccaaatcctt aaggaacacc cagttgaaaa cacccagctt cagaatgaga
    10141 agctctacct gtactacctg cagaacggca gggacatgta cgtggatcag gaactggaca
    10201 tcaatcggct ctccgactac gacgtggatc atatcgtgcc ccagtctttt ctcaaagatg
    10261 attctattga taataaagtg ttgacaagat ccgataaaaa tagagggaag agtgataacg
    10321 tcccctcaga agaagttgtc aagaaaatga aaaattattg gcggcagctg ctgaacgcca
    10381 aactgatcac acaacggaag ttcgataatc tgactaaggc tgaacgaggt ggcctgtctg
    10441 agttggataa agccggcttc atcaaaaggc agcttgttga gacacgccag atcaccaagc
    10501 acgtggccca aattctcgat tcacgcatga acaccaagta cgatgaaaat gacaaactga
    10561 ttcgagaggt gaaagttatt actctgaagt ctaagctggt ctcagatttc agaaaggact
    10621 ttcagtttta taaggtgaga gagatcaaca attaccacca tgcgcatgat gcctacctga
    10681 atgcagtggt aggcactgca cttatcaaaa aatatcccaa gcttgaatct gaatttgttt
    10741 acggagacta taaagtgtac gatgttagga aaatgatcgc aaagtctgag caggaaatag
    10801 gcaaggccac cgctaagtac ttcttttaca gcaatattat gaattttttc aagaccgaga
    10861 ttacactggc caatggagag attcggaagc gaccacttat cgaaacaaac ggagaaacag
    10921 gagaaatcgt gtgggacaag ggtagggatt tcgcgacagt ccggaaggtc ctgtccatgc
    10981 cgcaggtgaa catcgttaaa aagaccgaag tacagaccgg aggcttctcc aaggaaagta
    11041 tcctcccgaa aaggaacagc gacaagctga tcgcacgcaa aaaagattgg gaccccaaga
    11101 aatacggcgg attcgattct cctacagtcg cttacagtgt actggttgtg gccaaagtgg
    11161 agaaagggaa gtctaaaaaa ctcaaaagcg tcaaggaact gctgggcatc acaatcatgg
    11221 agcgatcaag cttcgaaaaa aaccccatcg actttctcga ggcgaaagga tataaagagg
    11281 tcaaaaaaga cctcatcatt aagcttccca agtactctct ctttgagctt gaaaacggcc
    11341 ggaaacgaat gctcgctagt gcgggcgagc tgcagaaagg taacgagctg gcactgccct
    11401 ctaaatacgt taatttcttg tatctggcca gccactatga aaagctcaaa gggtctcccg
    11461 aagataatga gcagaagcag ctgttcgtgg aacaacacaa acactacctt gatgagatca
    11521 tcgagcaaat aagcgaattc tccaaaagag tgatcctcgc cgacgctaac ctcgataagg
    11581 tgctttctgc ttacaataag cacagggata agcccatcag ggagcaggca gaaaacatta
    11641 tccacttgtt tactctgacc aacttgggcg cgcctgcagc cttcaagtac ttcgacacca
    11701 ccatagacag aaagcggtac acctctacaa aggaggtcct ggacgccaca ctgattcatc
    11761 agtcaattac ggggctctat gaaacaagaa tcgacctctc tcagctcggt ggagacagcg
    11821 gcggttccgg tggatccggc ggaagcacga acctgagtga tataattgaa aaggaaaccg
    11881 gtaaacagct cgttatccag gagtcgatcc tcatgctgcc cgaggaagtc gaggaagtca
    11941 tcggtaacaa accggaatcg gatattctgg tacatacggc ctatgatgaa tccaccgatg
    12001 agaatgtaat gttgttgaca tcggatgcac cggagtataa gccttgggcg ctcgtcatcc
    12061 aggactccaa tggcgagaac aagattaaga tgctctccgg aggatccaaa cgaacagctg
    12121 acggtagtga attcgaacca aaaaaaaagc gcaaggtata atctagagga tctttgtgaa
    12181 ggaaccttac ttctgtggtg tgacataatt ggacaaacta cctacagaga tttaaagctc
    12241 taaggtaaat ataaaatttt taagtgtata atgtgttaaa ctactgattc taattgtttg
    12301 tgtattttag attccaacct atggaactga tgaatgggag cagtggtgga atgcctttaa
    12361 tgaggaaaac ctgttttgct cagaagaaat gccatctagt gatgatgagg ctactgctga
    12421 ctctcaacat tctactcctc caaaaaagaa gagaaaggta gaagacccca aggactttcc
    12481 ttcagaattg ctaagttttt tgagtcatgc tgtgtttagt aatagaactc ttgcttgctt
    12541 tgctatttac accacaaagg aaaaagctgc actgctatac aagaaaatta tggaaaaata
    12601 tttgatgtat agtgccttga ctagagatca taatcagcca taccacattt gtagaggttt
    12661 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
    12721 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
    12781 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
    12841 tcaatgtatc ttatcatgtc tggatccact agtgtcgacg atgtaggtca cggtctcgaa
    12901 gccgcggtgc gggtgccagg gcgtgccctt gggctccccg ggcgcgtact ccacctcacc
    12961 catctggtcc atcatgatga acgggtcgag gtggcggtag ttgatcccgg cgaacgcgcg
    13021 gcgcaccggg aagccctcgc cctcgaaacc gctgggcgcg gtggtcacgg tgagcacggg
    13081 acgtgcgacg gcgtcggcgg gtgcggatac gcggggcagc gtcagcgggt tctcgacggt
    13141 cacggcgggc atgtcgacac tagttctagc cagcttttgt tccctttagt gagggttaat
    13201 ttcgagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
    13261 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
    13321 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
    13381 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
    13441 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
    13501 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
    13561 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
    13621 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
    13681 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
    13741 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
    13801 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
    13861 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
    13921 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
    13981 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
    14041 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
    14101 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
    14161 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
    14221 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
    14281 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
    14341 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
    14401 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
    14461 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
    14521 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
    14581 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
    14641 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
    14701 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
    14761 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
    14821 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
    14881 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
    14941 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
    15001 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
    15061 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
    15121 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
    15181 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
    15241 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
    15301 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
    15361 aaaagtgcca c
//
