Status
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Gene information
General information
TC001667 Gene

The sequence from the iB fragment is matched against the genes from the official gene set. The matche(s) is/are visualizied in a genome browser.

NC_007424.3: 2435420..2587325
Sequence information
Transcripts / Proteins (1) Get mRNA sequence
Get CDS sequence
Get protein sequence
Ortholog information (from OrthoDB)

The information about the homolog genes in Drosophila is provided by OrthoDB http://www.orthodb.org/. The homologs are sorted according to their similarity score. For an orthology statement additional analysis are required.
The data basis for the orthologs is OrthoDB v9 containing OGS3 for Tribolium castaneum and 2015_04 (r6.07) for Drosophila melanogaster.

Closest fly homologs
    TC001667
    iBeetle screen iB_03227

    Attention! iBeetle is a first pass screen. Hence, the documented phenotypes need to be confirmed by independent experiments using non-overlapping dsRNA fragments! Phenotypes with a penetrance > 50% are frequently reproduced. See Schmitt-Engel et al. 2015 for details on reproducibility.

    iB sequence
    Phenotype after pupal injection
    Usually 10 injected animals

    Pupal injection procedure

    Day 0: 10 female pupae of the pBA19 strain (muscle enhancer trap line) were injected with dsRNA.

    3 days post injection (dpi): Hatch control: Pupal and adult lethality as well as metamorphosis defects (molting, eclosion) were documented. For mating, 4 males of the black strain were added.

    9 dpi: First egg-lay was collected and incubated for cuticle analysis. Adult lethality and egg production (reduced/ no egg-lay) was documented.

    11 dpi: Second egg-lay was collected and incubated for embryonic muscle analysis. Adult lethality and egg production (reduced/ no egg-lay) were documented.

    Note: The adult morphology was not analyzed systematically at 3/9/11 dpi. Only obvious phenotypes, visible without magnification, have been annotated.

    13 dpi: Egg productivity and Ovary analysis
    The percentage of hatched larvae was documented and not hatched larvae/ eggs were embedded for cuticle analysis (15 dpi). In case of a reduction of egg production, 4 injected females were dissected to analyze the gross morphology of the ovaries.

    14 dpi: Analysis of embryonic musculature and early embryonic development
    Offspring of the injected females (hatched and not hatched larvae/ eggs) were analysed for embryonic lethality and muscle defects.

    15 dpi: Analysis of larval instar 1 cuticle
    Offspring of injected females were analysed and cuticle phenotypes were annotated.

    22 dpi: Stink gland analysis
    Documentation of defects in abdominal and thoracic stink glands (colour, size, content) of the injected femals.

    Terms used in the pupal injection procedure

    larval pantagmatic defects
    At least two tagmata (head, thorax, abdomen or terminus) show similar/ comparable defects.
    empty eggs
    Just empty shells are visible, no cuticle has been developed.
    strong defects
    L1 cuticle larvae which show severe defects. These strong defects are subdivided into three categories
    strongly fragmented (cuticle crumbs)
    No segments or appendages are identifiable, only residues of cuticle and bristles can be identified.
    cuticle remnants (with unidentifiable segments)
    Segments or parts of appendages are recognizable, but identity is not clear.
    cuticle remnants (with some identifiable segments)
    At least one segment or appendage is clearly regonizable.
    number of eggs on slide
    affected embryos usually (not always) do not hatch. Only the not hatched were used for cuticle analysis.
    For the embryonic muscle analysis the hatched as well as the not hatched were analysed.
    total number of affected eggs/embryos/larvae
    Summary of all different phenotypes/ phenotypic series which were annotated.
    inside-out
    The L1 larva shows a partial or complete inversion., e.g. bristles, appendages or parts of the abdominal segments are inverted into the interior of the cuticle.
    starved
    Dissected females show a strong resorption of fatbody predominately in the abdomen and the oogenesis is blocked. These are considered as eventually lethal phenotypes and the blocked oogenesis probably a secondary defect due to starvation.
    eclosion not fulfilled
    The emergence of the adult from the pupa stage is interrupted. This phenotype shows pupal as well as adult features.
    Please see the help page for more information
    Metamorphosis and survival

    Lethality 11 days after pupal injection: 30.0% (includes death as pupa, adult)

    eclosion not fulfilled (with pupal and adult features) - Number of animals showing the phenotype: 1 (3 dpi)
    Days post injection


    adult lethality - Number of animals showing the phenotype: 2 (9 dpi)
    Days post injection


    Analysis of embryonic musculature and early development
    number of eggs/embryos/larvae on slide: < 50 larval musculature degenerated - Penetrance (percentage of animals/eggs on slide): 50% - 80%

    larval musculature not present - Penetrance (percentage of animals/eggs on slide): less than 30%

    affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): more than 80%
    Analysis of larval stage 1 cuticle
    number of eggs/embryos/larvae on slide: > 50
    Phenotype after larval injection
    Usually 10 injected animals

    Larval injection procedure

    Day 0: 10 female L5-6 larvae of the D17Xhom strain (females express RFP in the eyes and in the CNS; pupae express GFP in the thorax) were injected with dsRNA.

    11 days post injection (dpi): Pupal morphology Documentation of larval/ pupal death and defects of metamorphosis (eclosion, delayed development). Screening for alterations of pupal head and appendages, pupal thorax, pupal thoracic musculature, pupal legs, pupal wings, pupal abdomen and pupal genital lobes.

    16 dpi: Adult morphology Documentation of larval/ pupal and adult death and defects of metamorphosis (eclosion, delayed development). Screening for alterations of adult head and appendages, adult thorax, adult elytra and hindwings, adult legs and cuticle properties. For mating 4 males of the black strain were added.

    19 dpi: Sieving and egg-lay Documentation of adult lethality and enduring metamorphosis defects.

    22 dpi: Ovary analysis (Fertility) In case of a reduction of egg production 4 females were dissected and the gross morphology of the ovaries were analysed.

    41 dpi: Fertility and stink gland analysis Documentation of defects in abdominal and thoracic stink glands development (colour, size, content) of the injected females. Check for fertility of injected larvae (no or decreased offspring).

    Terms used in the larval injection procedure

    pupal/ adult pantagmatic defects
    At least two tagmata (head, thorax, abdomen or terminus) show similar/ comparable defects.
    pupal molt not fulfilled
    The hatching from the larva to the pupa is interrupted. Either it dies as prepupa or the phenotype shows larval as well as pupal features.
    eclosion not fulfilled
    The emergence of the adult from the pupa stage is interrupted. This phenotype shows pupal as well as adult features.
    starved
    Dissected females show a strong resorption of fatbody predominately in the abdomen and the oogenesis is blocked. These are considered as eventually lethal phenotypes and the blocked oogenesis probably a secondary defect due to starvation.
    Metamorphosis and survival

    Lethalities 11 days after larval injection: % (includes death as larva, prepupa, pupa)
    Lethalities 22 days after larval injection: % (includes death as larva, prepupa, pupa, adult)

    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC001667 RA
      MCPCELCVL CFFVLCVYT VTAFNVDSF NYATYQVPS 
      YDGHDSMFG FSIALHRER NTGWLIVGA PEASSSQPG 
      VIHGGAVYK CPTDRDGGC EEIPFDTKG NNRDASNNL 
      MDNKTEQWF GATVSSSGR AEGPIVACA PRYTWFPRG 
      RSRRDPVGT CYVCNGFFE KFNEYSPCR TYQWGYHRQ 
      GSCQAGFSA AINSVGDRL FIGSPGSWY WQGQMYSVD 
      AHATFEFKP TLFSATYGD EGQVHQQSL ETRPAVFAT 
      TEGTEKDDD SYMGYSTIV GDFLGTGEE GIAVGMPRG 
      NNLKGKVLL FTMNLTNYV QHFTSDQIG SYYGYALAA 
      CDVDGDGKL DLIVGAPMY TVQNNKGKY DVGRVYVIY 
      HESDRGRFV ANYVIDGFN SKSRFGQAV ASLGDINQD 
      GFGDFAVGA PYDGAGNRG AVYIYHGSS KGVRKKFSQ 
      VIFAEDVAP SLTTFGFSI TGGMDLDNN QYPDMAVGA 
      YESNKAFFF RARPVISVE ASVKFLTHN KFIDITRPD 
      YTLRSDGTT ATKTQIQFC SRYYGKGIP NSLNLGVEY 
      ILDSKKLIN SRMGFVDDE GQSRKNATI TMLKGQDMK 
      CTNPIDVYV KSEIRDKLT ALVVEARYW MQSDQDLTP 
      SNLQQLPRN PRSFLTPVL DLNSPPSKK DSISIQKNC 
      GSDNICIPD LVISVTPSV QEYYLDSGE NFYLNISVT 
      NNGEDAFES TVEVKYPDG FFYISSQDI QVVSHVLCS 
      ASENFTIKC DIGNPVPSN KVVNFKLLW QPNYAKELP 
      SSLFFEVLV NSTNEDLPE TRYNNKQKI GISILYDVI 
      QEIRGWSIP NEVLFTPSS SLYAADISN QTHRTSPEN 
      RLSEDVIGP QVIHVYELK NGGRTTIKA TEIFFVWPA 
      TTASGEDFL YLVDPPHFS FTENLNLKC GPIPANYRD 
      FEVPTRRKS IWELYDIDV STDTTSEKY MAGKNQTYH 
      KIGSTIGGK IVTSDDKIN KEVEKSGDS SKVFDVRHN 
      KTTWHSSSR VQIFEETKR LTGNFRRFR LGGKQYAYR 
      CLQSDQEVD CGTDFTFSR LDLSKLLQT SSAGPNRYS 
      CLDQDSFGN WRKTDCGHV PTIPVIPTV TEGETRVQI 
      FGANDQLVN QLESRSYKK QTIDGKDFI YRCFRRANG 
      REEEIDCGQ NFSFSSIDL PWILEQSGI QQQEGNVRY 
      ACLKPDSDG NLRSTTCDN VPKIRFKTQ IIRKFWGSN 
      DTHYWNETG VYDLRGSST IRVQVFQGD DDLRRQLQS 
      FYHLQHLRG YNFIYRCLD GNEEVDCDD FSLAQFTDF 
      TDFGVLLTT GVQGDGKIR YICLQSDNY HNLRSVPCG 
      HLPPIHFKT QVIQTSWKS NGTHYWNNT GVYRVSKKY 
      DFSTVQVFQ GGSEMAEQL GGTYRFEKV DGQVFIYRC 
      FKNTFEVDC GQFNLLSLD LLQLMRTVA IQREGNLRY 
      FCLEMDSFG NWRGADCGH LPRIHVTEP KRVITQEHI 
      LKRCLKLVN GNWQELDCR AGFHSNDGT FIEECYQTI 
      NGRRDRINC TQVPPGSTF DYVQTNTRY NQESASTSH 
      GTDYFSGHG RQTYRPSYG TDYNYNQGQ AFGNQSYSY 
      ENSYETELY EDEDDKAIS GYMNPHDVA PTERQFGRR 
      DPKTYGTYG EARSAGVAA GGLGEAAQG RGFTAVALD 
      LGNLGSGDS HSSHWSETH HVAPVHNTI GRDNVPTDA 
      PSPRNWHHG RKKRQIIGD SHPSIAGIK EEFPCKNLG 
      CVYMRCTIE NLQHGKPLT IALRGRLNA KVLRDMKLD 
      NALQFSSMM LARVHKVPH LVSLKGSKP VVKEVETAV 
      KTPEEIKSE TVPLWVVVL SAVAGTIIL LLIIFLLYK 
      LGFFKRNRP SSAPERQPL NRNGYHHGD EAL
    CDS sequence
    • >TC001667 RA
      atgtgtccg tgcgaattg tgtgtgttg tgttttttc 
      gtgttatgt gtatacaca gtgaccgct tttaatgtc 
      gatagtttt aattacgct acgtatcaa gtgcccagt 
      tacgatggt catgattcc atgttcgga ttcagcata 
      gctttacat cgagaaaga aacactgga tggttgatt 
      gtgggggca ccggaagct tcatctagt caacctggg 
      gttattcac ggaggtgct gtctacaag tgtccaacg 
      gatagagat gggggctgc gaggaaata cctttcgac 
      actaaaggc aacaatcgg gacgccagc aacaaccta 
      atggacaac aaaacggag caatggttc ggtgcaacc 
      gtcagttct tcaggccgt gcggaaggt cctatcgtg 
      gcatgtgct cctcgttac acctggttt ccacgcggt 
      cgttcccgt cgagatcct gtaggaacc tgctacgtt 
      tgcaacggt ttcttcgag aaattcaac gaatactca 
      ccttgtaga acatatcaa tggggatac catcggcaa 
      gggtcatgt caggctggt ttcagcgca gccataaac 
      tcggtcgga gaccgcctt ttcatcggt tcgccgggc 
      agttggtat tggcaagga caaatgtat tccgtcgat 
      gctcacgcc acgtttgaa tttaaacct acgctattc 
      tcagcgacc tacggcgat gaaggacaa gtccaccaa 
      cagagtctc gaaacacgt ccggcagtt ttcgccacc 
      actgaaggt accgaaaaa gacgacgac tcctacatg 
      ggttattcg accatcgtg ggcgacttt ctgggcact 
      ggggaagaa ggcatcgct gtgggtatg cccagagga 
      aacaatctc aagggaaaa gtgctcctt ttcaccatg 
      aatcttacc aattacgtc caacacttc accagtgat 
      caaatcggg tcgtactat ggctacgcc ctcgcagcg 
      tgtgatgtc gatggtgat ggtaaactg gatttgatc 
      gtgggggca cccatgtac acagtgcag aataataaa 
      gggaaatat gacgtgggg agggtctat gtcatctac 
      catgaaagc gataggggt cgtttcgtg gccaactat 
      gtgatcgat gggttcaat tccaagtca agatttggc 
      caagctgtg gcttcgctt ggtgatatc aatcaagac 
      ggttttgga gactttgca gtgggggct ccttatgat 
      ggtgccgga aaccgtggt gctgtctac atttatcat 
      ggctcaagc aaaggcgtc cgcaagaag ttctcgcaa 
      gttattttt gcagaagat gtggctccg tctttaaca 
      acttttgga ttttccata actggagga atggatctt 
      gacaataac caatatcca gacatggct gttggtgct 
      tacgaatcc aacaaagct ttcttcttc agagctcgt 
      ccagtgatc agcgttgaa gcttcagtc aaatttctt 
      acacataat aaattcatt gatatcact aggcctgat 
      tatacgctt cgaagtgat ggtactacg gctaccaag 
      acccaaatt cagttctgt agcaggtac tatggaaaa 
      gggattccg aattcttta aatttgggt gttgagtac 
      attttggat tcaaaaaaa ttgatcaat tcgcggatg 
      ggtttcgtg gatgatgaa ggccaaagt aggaaaaat 
      gctacaata actatgttg aagggacaa gatatgaag 
      tgtacaaac ccgatcgat gtttatgtg aagagcgaa 
      attcgcgac aaactcaca gccctagtc gttgaagcc 
      cgttactgg atgcaaagc gaccaagac ctcacaccc 
      tcaaacctc caacaactg ccccgcaac ccccgatcc 
      ttcctcacc ccagtcctc gacctcaac tcaccccca 
      tccaagaaa gactcaatc agcatacag aagaactgc 
      ggttcggac aacatctgt atcccagac ttggtcatt 
      tcagtcact ccgagcgtc caggagtac tacctggat 
      tcgggcgaa aacttctac ttgaacatt tcagtcaca 
      aacaacgga gaagacgca tttgaatca accgtcgaa 
      gttaaatac cccgacgga tttttctac atttcgagc 
      caagacatc caagtggta tcgcacgtg ttgtgcagt 
      gcctctgaa aatttcacc atcaagtgt gatattggt 
      aatccggtg ccgtcgaat aaagtcgtc aatttcaaa 
      ctattgtgg caaccgaac tacgccaaa gaactaccc 
      tcgagtcta tttttcgag gttttggtc aacagtacc 
      aacgaagac ttgcccgaa actcgctac aacaacaag 
      caaaagatt ggaatttca attctctac gacgttatt 
      caagaaatt cggggctgg tcgatacct aacgaagtg 
      ttattcacg cccagttcg tcactttac gctgccgat 
      atctccaat caaacgcat cgcacttcg cctgaaaac 
      cgcctctct gaggacgtt atcggcccc caggtcatc 
      cacgtttac gaattgaaa aacggcgga cgaactacg 
      atcaaagcc accgaaata tttttcgtt tggccggcg 
      accacagca agtggcgaa gatttccta tacttggtc 
      gatcccccc cacttttct tttacggaa aatttgaac 
      ctcaaatgt ggcccaatc cccgccaat taccgcgat 
      ttcgaagtc ccaacacgg agaaaatca atctgggaa 
      ctgtacgac attgacgtt tccaccgac acaacatcc 
      gagaaatac atggcgggg aaaaaccaa acgtaccac 
      aagatcggt tcgacaatt ggcggcaag attgtaaca 
      agcgacgac aaaatcaac aaagaggtg gaaaaatcg 
      ggcgatagt tccaaagtt ttcgacgta cgccacaac 
      aaaaccact tggcatagc tcgtcccgg gtgcaaatt 
      ttcgaagaa acgaagcgt ctgactgga aatttccga 
      cgtttccgt cttgggggc aaacaatac gcctatcgt 
      tgccttcag tcggaccag gaggttgat tgtgggacc 
      gatttcacg ttctcacgt cttgactta tccaagctg 
      ttgcaaact tcaagtgct ggaccgaat cgctactct 
      tgcctcgac caggattcg ttcggcaac tggcgcaag 
      accgactgt ggacacgtc cccacaatc cctgttatt 
      cccaccgtg acggaaggc gaaacacgt gtacaaatt 
      ttcggggcg aacgaccaa ctggtcaac cagttagag 
      agccggagt tacaaaaaa cagacaatt gacgggaag 
      gatttcatc tatcggtgt ttccggcgg gctaacggg 
      cgggaagag gagatcgat tgcgggcaa aacttctcg 
      ttttcaagc attgacctg ccctggatt ctggaacag 
      tcgggaata caacaacaa gaagggaac gttcggtac 
      gcgtgcctt aaacccgat tctgacggc aatctccga 
      agtaccaca tgcgataat gtccccaaa attcgtttc 
      aagacgcaa attatacga aagttttgg ggttccaat 
      gacacccac tactggaac gagacgggg gtttacgac 
      ctgaggggg agctcgaca attcgggtg caagtcttt 
      caaggggat gacgattta aggcgacaa ttgcaaagc 
      ttttatcat ctgcaacac ctccgaggg tacaatttc 
      atctaccgt tgtttggac ggaaacgaa gaggttgac 
      tgtgatgac ttttcccta gcacagttc accgatttc 
      accgatttt ggggttttg ctcaccaca ggtgtgcaa 
      ggggatggt aaaatacgg tatatctgt ctccaaagt 
      gacaattac cacaactta cggagcgtc ccttgtggg 
      caccttccc ccaattcac ttcaaaaca caagtgatt 
      caaacatcg tggaaatca aacggtact cactattgg 
      aacaatacg ggagtctac cgagtgtca aaaaagtac 
      gactttagc acagttcag gtgttccaa gggggtagc 
      gaaatggcc gagcaattg ggtggtact taccgcttt 
      gaaaaagtg gacggtcaa gtgtttatc tatcgctgt 
      ttcaaaaac acgtttgag gttgactgt ggccaattc 
      aatttactc agtttggat ttgttgcaa ttgatgcga 
      accgttgcg attcaacgt gaaggcaac cttcggtac 
      ttttgtctg gaaatggac agttttggt aactggagg 
      ggggccgac tgtggccat ttgccccga attcacgtc 
      accgaaccc aaaagagtg atcacacaa gaacacatc 
      ttgaaacgc tgtctgaaa cttgttaat ggcaattgg 
      caagagttg gattgtcgt gccggtttt cactcaaac 
      gatggtacc tttattgaa gagtgttac cagacaatc 
      aacggtcga agagacaga attaattgt acgcaagta 
      ccaccaggg agcactttt gactacgtc caaaccaac 
      acacgctac aaccaagaa agtgcaagt actagtcat 
      ggtactgat tatttctct ggtcatggg cgccaaacc 
      tatcgtccc tcctacgga acggactac aactacaac 
      caaggtcaa gcctttggt aaccaatcc tacagttac 
      gaaaacagc tacgagacc gaactctac gaagacgaa 
      gatgataag gcgatttcg ggttatatg aacccgcac 
      gatgtcgca ccgaccgaa cgccaattc ggccgaaga 
      gacccgaag acgtacgga acgtacggc gaggccagg 
      agtgcagga gttgcagct gggggtctt ggtgaggcg 
      gcccaaggt cggggtttc accgcagtt gcgctagat 
      ttgggtaat ttgggatcg ggtgattcc cattcctcc 
      cattggtct gaaacccat catgtggca ccggtgcat 
      aataccata gggcgagat aatgtacca acggatgct 
      ccaagtccc aggaactgg caccatggg aggaaaaaa 
      cgacagatt attggtgat agtcatccg tctatcgct 
      gggattaaa gaagagttc ccttgcaag aatcttggg 
      tgtgtctat atgagatgc actattgag aatttgcaa 
      catgggaaa ccactgacg attgcgctg agggggcga 
      ttgaacgca aaagtgttg agagatatg aaactagac 
      aacgctttg caattttcg tccatgatg ttggcccgc 
      gtgcacaaa gtgccccat ctggtgtcg cttaaaggc 
      agtaaacca gtcgtgaaa gaagtggaa actgctgtc 
      aaaaccccc gaagaaatc aagtccgaa actgtgcct 
      ttgtgggtc gtcgtactg tcagcggtc gctggcacg 
      atcatcctc ctcctcatc attttccta ctttataaa 
      ctcggtttc ttcaaacga aaccggcct tcgagcgcc 
      cccgagcgg cagcccctc aaccgcaac ggctatcac 
      catggcgac gaagctctg tga
    mRNA sequence
    • >TC001667 RA
      tatatgtgt caaatctca accctctag aaaatctct 
      tcaacccaa tggaaagca ccttcaagt ttactcact 
      atggtgtaa ggaaatatt ccttgcttg agaaggacc 
      tgtgtaaca gcgctgttt tcatttttt cgatctttt 
      ctgtttgct ccaacattg agatgttca aacgtttgt 
      cagtcgcgc atcaacttg ggtaatgta aagagcgcg 
      aagtataat ttgaaacat gtgcactgc cctcacgaa 
      ccatttcta gcaccgacc aatttttag aatgaaatt 
      tcccacttt tatcatttt ttgaactct ccttaccct 
      ctaaactaa ataaataaa aaaaatgtg tccgtgcga 
      attgtgtgt gttgtgttt tttcgtgtt atgtgtata 
      cacagtgac cgcttttaa tgtcgatag ttttaatta 
      cgctacgta tcaagtgcc cagttacga tggtcatga 
      ttccatgtt cggattcag catagcttt acatcgaga 
      aagaaacac tggatggtt gattgtggg ggcaccgga 
      agcttcatc tagtcaacc tggggttat tcacggagg 
      tgctgtcta caagtgtcc aacggatag agatggggg 
      ctgcgagga aataccttt cgacactaa aggcaacaa 
      tcgggacgc cagcaacaa cctaatgga caacaaaac 
      ggagcaatg gttcggtgc aaccgtcag ttcttcagg 
      ccgtgcgga aggtcctat cgtggcatg tgctcctcg 
      ttacacctg gtttccacg cggtcgttc ccgtcgaga 
      tcctgtagg aacctgcta cgtttgcaa cggtttctt 
      cgagaaatt caacgaata ctcaccttg tagaacata 
      tcaatgggg ataccatcg gcaagggtc atgtcaggc 
      tggtttcag cgcagccat aaactcggt cggagaccg 
      ccttttcat cggttcgcc gggcagttg gtattggca 
      aggacaaat gtattccgt cgatgctca cgccacgtt 
      tgaatttaa acctacgct attctcagc gacctacgg 
      cgatgaagg acaagtcca ccaacagag tctcgaaac 
      acgtccggc agttttcgc caccactga aggtaccga 
      aaaagacga cgactccta catgggtta ttcgaccat 
      cgtgggcga ctttctggg cactgggga agaaggcat 
      cgctgtggg tatgcccag aggaaacaa tctcaaggg 
      aaaagtgct ccttttcac catgaatct taccaatta 
      cgtccaaca cttcaccag tgatcaaat cgggtcgta 
      ctatggcta cgccctcgc agcgtgtga tgtcgatgg 
      tgatggtaa actggattt gatcgtggg ggcacccat 
      gtacacagt gcagaataa taaagggaa atatgacgt 
      ggggagggt ctatgtcat ctaccatga aagcgatag 
      gggtcgttt cgtggccaa ctatgtgat cgatgggtt 
      caattccaa gtcaagatt tggccaagc tgtggcttc 
      gcttggtga tatcaatca agacggttt tggagactt 
      tgcagtggg ggctcctta tgatggtgc cggaaaccg 
      tggtgctgt ctacattta tcatggctc aagcaaagg 
      cgtccgcaa gaagttctc gcaagttat ttttgcaga 
      agatgtggc tccgtcttt aacaacttt tggattttc 
      cataactgg aggaatgga tcttgacaa taaccaata 
      tccagacat ggctgttgg tgcttacga atccaacaa 
      agctttctt cttcagagc tcgtccagt gatcagcgt 
      tgaagcttc agtcaaatt tcttacaca taataaatt 
      cattgatat cactaggcc tgattatac gcttcgaag 
      tgatggtac tacggctac caagaccca aattcagtt 
      ctgtagcag gtactatgg aaaagggat tccgaattc 
      tttaaattt gggtgttga gtacatttt ggattcaaa 
      aaaattgat caattcgcg gatgggttt cgtggatga 
      tgaaggcca aagtaggaa aaatgctac aataactat 
      gttgaaggg acaagatat gaagtgtac aaacccgat 
      cgatgttta tgtgaagag cgaaattcg cgacaaact 
      cacagccct agtcgttga agcccgtta ctggatgca 
      aagcgacca agacctcac accctcaaa cctccaaca 
      actgccccg caacccccg atccttcct caccccagt 
      cctcgacct caactcacc cccatccaa gaaagactc 
      aatcagcat acagaagaa ctgcggttc ggacaacat 
      ctgtatccc agacttggt catttcagt cactccgag 
      cgtccagga gtactacct ggattcggg cgaaaactt 
      ctacttgaa catttcagt cacaaacaa cggagaaga 
      cgcatttga atcaaccgt cgaagttaa ataccccga 
      cggattttt ctacatttc gagccaaga catccaagt 
      ggtatcgca cgtgttgtg cagtgcctc tgaaaattt 
      caccatcaa gtgtgatat tggtaatcc ggtgccgtc 
      gaataaagt cgtcaattt caaactatt gtggcaacc 
      gaactacgc caaagaact accctcgag tctattttt 
      cgaggtttt ggtcaacag taccaacga agacttgcc 
      cgaaactcg ctacaacaa caagcaaaa gattggaat 
      ttcaattct ctacgacgt tattcaaga aattcgggg 
      ctggtcgat acctaacga agtgttatt cacgcccag 
      ttcgtcact ttacgctgc cgatatctc caatcaaac 
      gcatcgcac ttcgcctga aaaccgcct ctctgagga 
      cgttatcgg cccccaggt catccacgt ttacgaatt 
      gaaaaacgg cggacgaac tacgatcaa agccaccga 
      aatattttt cgtttggcc ggcgaccac agcaagtgg 
      cgaagattt cctatactt ggtcgatcc cccccactt 
      ttcttttac ggaaaattt gaacctcaa atgtggccc 
      aatccccgc caattaccg cgatttcga agtcccaac 
      acggagaaa atcaatctg ggaactgta cgacattga 
      cgtttccac cgacacaac atccgagaa atacatggc 
      ggggaaaaa ccaaacgta ccacaagat cggttcgac 
      aattggcgg caagattgt aacaagcga cgacaaaat 
      caacaaaga ggtggaaaa atcgggcga tagttccaa 
      agttttcga cgtacgcca caacaaaac cacttggca 
      tagctcgtc ccgggtgca aattttcga agaaacgaa 
      gcgtctgac tggaaattt ccgacgttt ccgtcttgg 
      gggcaaaca atacgccta tcgttgcct tcagtcgga 
      ccaggaggt tgattgtgg gaccgattt cacgttctc 
      acgtcttga cttatccaa gctgttgca aacttcaag 
      tgctggacc gaatcgcta ctcttgcct cgaccagga 
      ttcgttcgg caactggcg caagaccga ctgtggaca 
      cgtccccac aatccctgt tattcccac cgtgacgga 
      aggcgaaac acgtgtaca aattttcgg ggcgaacga 
      ccaactggt caaccagtt agagagccg gagttacaa 
      aaaacagac aattgacgg gaaggattt catctatcg 
      gtgtttccg gcgggctaa cgggcggga agaggagat 
      cgattgcgg gcaaaactt ctcgttttc aagcattga 
      cctgccctg gattctgga acagtcggg aatacaaca 
      acaagaagg gaacgttcg gtacgcgtg ccttaaacc 
      cgattctga cggcaatct ccgaagtac cacatgcga 
      taatgtccc caaaattcg tttcaagac gcaaattat 
      acgaaagtt ttggggttc caatgacac ccactactg 
      gaacgagac gggggttta cgacctgag ggggagctc 
      gacaattcg ggtgcaagt ctttcaagg ggatgacga 
      tttaaggcg acaattgca aagctttta tcatctgca 
      acacctccg agggtacaa tttcatcta ccgttgttt 
      ggacggaaa cgaagaggt tgactgtga tgacttttc 
      cctagcaca gttcaccga tttcaccga ttttggggt 
      tttgctcac cacaggtgt gcaagggga tggtaaaat 
      acggtatat ctgtctcca aagtgacaa ttaccacaa 
      cttacggag cgtcccttg tgggcacct tcccccaat 
      tcacttcaa aacacaagt gattcaaac atcgtggaa 
      atcaaacgg tactcacta ttggaacaa tacgggagt 
      ctaccgagt gtcaaaaaa gtacgactt tagcacagt 
      tcaggtgtt ccaaggggg tagcgaaat ggccgagca 
      attgggtgg tacttaccg ctttgaaaa agtggacgg 
      tcaagtgtt tatctatcg ctgtttcaa aaacacgtt 
      tgaggttga ctgtggcca attcaattt actcagttt 
      ggatttgtt gcaattgat gcgaaccgt tgcgattca 
      acgtgaagg caaccttcg gtacttttg tctggaaat 
      ggacagttt tggtaactg gaggggggc cgactgtgg 
      ccatttgcc ccgaattca cgtcaccga acccaaaag 
      agtgatcac acaagaaca catcttgaa acgctgtct 
      gaaacttgt taatggcaa ttggcaaga gttggattg 
      tcgtgccgg ttttcactc aaacgatgg tacctttat 
      tgaagagtg ttaccagac aatcaacgg tcgaagaga 
      cagaattaa ttgtacgca agtaccacc agggagcac 
      ttttgacta cgtccaaac caacacacg ctacaacca 
      agaaagtgc aagtactag tcatggtac tgattattt 
      ctctggtca tgggcgcca aacctatcg tccctccta 
      cggaacgga ctacaacta caaccaagg tcaagcctt 
      tggtaacca atcctacag ttacgaaaa cagctacga 
      gaccgaact ctacgaaga cgaagatga taaggcgat 
      ttcgggtta tatgaaccc gcacgatgt cgcaccgac 
      cgaacgcca attcggccg aagagaccc gaagacgta 
      cggaacgta cggcgaggc caggagtgc aggagttgc 
      agctggggg tcttggtga ggcggccca aggtcgggg 
      tttcaccgc agttgcgct agatttggg taatttggg 
      atcgggtga ttcccattc ctcccattg gtctgaaac 
      ccatcatgt ggcaccggt gcataatac catagggcg 
      agataatgt accaacgga tgctccaag tcccaggaa 
      ctggcacca tgggaggaa aaaacgaca gattattgg 
      tgatagtca tccgtctat cgctgggat taaagaaga 
      gttcccttg caagaatct tgggtgtgt ctatatgag 
      atgcactat tgagaattt gcaacatgg gaaaccact 
      gacgattgc gctgagggg gcgattgaa cgcaaaagt 
      gttgagaga tatgaaact agacaacgc tttgcaatt 
      ttcgtccat gatgttggc ccgcgtgca caaagtgcc 
      ccatctggt gtcgcttaa aggcagtaa accagtcgt 
      gaaagaagt ggaaactgc tgtcaaaac ccccgaaga 
      aatcaagtc cgaaactgt gcctttgtg ggtcgtcgt 
      actgtcagc ggtcgctgg cacgatcat cctcctcct 
      catcatttt cctacttta taaactcgg tttcttcaa 
      acgaaaccg gccttcgag cgcccccga gcggcagcc 
      cctcaaccg caacggcta tcaccatgg cgacgaagc 
      tctgtgatt ccccggtag ctacattac tgcctaaac 
      caaaagact tccacttaa cttattgtc ctcgtgtga 
      atgtatagt tgaaatgac cgaaacgcc atctgtatt 
      tacgaagtc ataacagac taattttaa ttaatactt 
      actaaaact ctgtttagt gccttattc actagtgcc 
      actttgcaa actgttgca agagaaact gtttgttat 
      gataggaat taatttagt cgtatcgaa cgatcaatg 
      tttgtattg tacttagtt acaatttgt tgttttgcc 
      actaattaa tttaattaa aacgaagaa gaaacgatt 
      tgttgccgt cgaaatttg tacattgta atttgacct 
      tgtaaataa aactattga ttatgttag atgattctt 
      ttatttttt gtactgggt acgttaaaa aaatgcgat 
      acaaaaata atgttagtg aagtaaaat taattgcac 
      atacaaata aacatcgta tggtacatt atttgtact 
      taaaacatg gacgtggca acactaacg ccacaattt 
      tttctttaa atttcaaca acaatcaga ttatatttc 
      cagatctga tcaatgtaa aacaatcac aatgacgtt 
      atttctgct ataatcgat aattttatc tttttttaa 
      attcctaga ttagataat tatcacaag tattcaacg 
      tctaggctt ggcaatgtt ccttttggt gattcaatt 
      acataccac aaataagat ttataaata cataaaatt 
      ttgttattt taattaatt atttaatcg gttactcaa 
      ttggtaatc catcaaaac cagacgatt tggttcgtt 
      tctaagcaa gaagatcaa aatatttca acaaaacaa 
      tgccaaaat aatattccg caataaaaa ttacgtttc 
      gaa
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