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Gene information
General information
TC003226 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.

scaffold_seqid13: 94939..101560
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
    TC003226
    iBeetle screen iB_03522
    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: 10.0% (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


    Fertility
    ovary surface rough - Number of animals showing the phenotype: 4
    egg chamber completenot present - Number of animals showing the phenotype: 4
    vitellarium shortened - Number of animals showing the phenotype: 2
    vitellarium potentially fused - Number of animals showing the phenotype: 1

    tropharium partially structure irregular - Number of animals showing the phenotype: 1
    Analysis of larval stage 1 cuticle
    number of eggs/embryos/larvae on slide: < 20
    egg without cuticle embryo/egg without visible larval cuticle in cuticle preparation - Penetrance (percentage of animals/eggs on slide): 50% - 80%

    affected eggs/embryos/larvae total percentage of affected eggs/embryos/larvae on slide - Penetrance (percentage of animals/eggs on slide): 50% - 80%
    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: 0.0% (includes death as larva, prepupa, pupa)
    Lethalities 22 days after larval injection: 90.0% (includes death as larva, prepupa, pupa, adult)

    adult lethality immediately after eclosion - Number of animals showing the phenotype: 2 (16 dpi)
    Days post injection


    eclosion not fulfilled (died as pupa) - Number of animals showing the phenotype: 1 (16 dpi)
    Days post injection


    eclosion not fulfilled (with pupal and adult features) - Number of animals showing the phenotype: 2 (16 dpi)
    Days post injection



    eclosion not fulfilled (not completely eclosed) - Number of animals showing the phenotype: 3 (16 dpi)
    Days post injection



    adult lethality - Number of animals showing the phenotype: 1 (19 dpi)
    Days post injection


    Fertility
    fertility after larval injection decreased (no living offspring) -
    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC003226 RA
      MNKNLQSPT KLTEFAPIQ TEEKPQTVG QFITKIFKF 
      NKGPNSEEQ NSGLTPNSD NVSQESLPT WAIESDSSA 
      TPFHVDVNE GRSLPNVLK RISNLLALK TTNLQDYSD 
      TELKQYWMP DSVSKECYQ CCEKFTTFR RRHHCRVCG 
      QIFCSQCCN QQIPGKIFG CTGDLRVCT YCCKVVLSY 
      LQSSDVGAD LTADLKLLQ ENLQSKFGN DVLSPTVQD 
      NIQNLNKDS VEVGNILRR KISVGYQEE KFASGQSSM 
      YLTTEEKCK ALQNSVSLR ALFEEICRT TTGIPLGTH 
      RYRLRTYTD CFLGSELVD WLICQQKAN SRVQAAAIS 
      QALLEGGYI ECVSDPCSF VDGYALYKP GIFVTPEVL 
      NHNYFEVPN QEEPIWVQQ IPQESSTTD SDNEQVSVK 
      KRGPLTSSS SYMLDLNVE ANTVYLSRP LASSYSIQS 
      GDSGDVSCC ETDITTVRT SEQREFVPE AGWHNASNL 
      REENGEKLA YNLLTEAYQ QHEQGLLKQ LLASNGLLL 
      SWADVIIPL CNEIINVIR PDKNHDAED LDIRHYIKF 
      KKLSGGSRT DTKLISGIV FTKNVAHKG MLTEIDNPK 
      ILLLQCSIV YQRTEGRLM SLEPVLMQE HEYLRHVAA 
      RIVALQPDI VLVHRNVSR LAQDLLRQH GITLVHNVK 
      QSVLDQLAR CTEADLVTA VDAHIGRPR LGTCKKFYL 
      KTYNVDKGG AKTLMFFEG LPMVHLGGT VLLRGGSRQ 
      ELTRLEKVV SFCLFASYN WRLEKSFLM DEFAQPPNP 
      NCEFLEDSS KESSPRLQT VVKYDSGGK KMNSEMVED 
      FTDPLQSSF VESMTEGKE ILAVAELPF SNNFRKSLD 
      DSILCISPY LMFSVPYLE TEVGKKCKL RKFFPTEIY 
      FSEQFTNQT KVKIIKDIG EDVKENEQK SVKPLHPFL 
      RTKITTSVD NNEIQSLLA HFRACGGSY EKREILCKP 
      PVQVDEEIL QNKDKNYYN KLDVLEINN HQRLSVLFC 
      SFSHESTNS PAFCVNPWI VNMNFYGSN DIPLGCFLE 
      RYCFRSTYN CPSKPCGTP MFKHIRRFV HNAGCVSIS 
      LNNFDNEFT EENIVMWSW CTKCQSVSP VVLMSADTW 
      SFSFAKYLE LKFYGGLYS RRGNTPCGH SLHHDHYQY 
      FGYKNSVAS FKYTPIQVW DISLPPPVI YIQYDIEKQ 
      QTELIDEIR TMAQKGHEI YSLILEKLS CIPAELEGL 
      GNLKQLLLK EQTQFKQKV EEVQLKLTS PTIENKQFD 
      ETPEKLHIA YWKISDSLI RIKRLIVET VDNWNLRLS 
      ESARKRDDK KKDRTSYSD LESPTIPEN KTLSETDIS 
      HSETSNSAT ITKKIKSLD QSDESELSM SSSPKCHHR 
      SQSDGTVMS HNEEQNDCK KDSDKKTVK NILSQLLPS 
      SSALTLIPN PFNPQDHYT LPTGVSVPI IVYESEPSS 
      IIAYALNSY DYKKSFEDL TKKSNTEQT PSPIVKRKN 
      PNTDKNDET SGLLGFLRN KNDLNSPVS ASESPQNIE 
      VTEKSKNLH IEVQFQDTH CNFFCRTYF AEKFASLRG 
      LVLPIGEEG YVRSLARSV QWNARGGKS GSNFAKTAD 
      DRFVLKEMS KSEVQLFLE SAPNYFNYM QKCYGTGQP 
      TLLGKIIGI YQIIFKNNN SNVTLRTNL LVMENLFYK 
      RTVSQKFDL KGSMRNRLV VPDNQEGEI VLLDENLLK 
      MTCDAPLYI LPHSKAVLT AAIQNDTEF LSAQSVMDY 
      SLLVGLDSE NRELVLGII DYIRTFTWD KKLETMVKK 
      SGILGGQGK LPTIVSPEE YQKRFIEAM HRYFLEVPD 
      HWAGLGKGL EF
    CDS sequence
    • >TC003226 RA
      atgaacaag aatctacaa tcccccacg aaattaact 
      gaattcgcg cccattcaa acggaagaa aagccccaa 
      actgtgggt cagttcatt accaagatt ttcaaattt 
      aataaaggc ccaaattcg gaagaacaa aactcgggt 
      ttaaccccg aattcggat aatgtgtct caggagtct 
      ctaccgaca tgggcaatc gagtcagac tcgtctgca 
      acccctttt cacgtagat gtgaacgag gggcgaagc 
      cttcccaat gttttgaaa cgaatcagt aatttacta 
      gcattaaaa accacgaac ttgcaagac tattcagac 
      accgaactg aaacaatat tggatgccg gattctgtg 
      agcaaagaa tgctaccaa tgttgcgaa aaattcact 
      acgttcaga cgccgacat cactgtcgt gtctgtggc 
      caaattttt tgctcacag tgttgcaat caacaaatt 
      ccgggaaaa attttcgga tgtactggt gatttaagg 
      gtttgcaca tattgctgc aaagtggtg ttatcatat 
      ctacagtcg tcagatgtt ggggctgac ctaactgcc 
      gatttaaag ctcctccaa gagaattta caaagcaaa 
      ttcggcaac gatgttctc tccccaact gtgcaagac 
      aacatacaa aatttgaac aaggattca gtcgaagtt 
      gggaatatt ttacgccgg aaaatttcc gtggggtat 
      caggaggag aaattcgca tctggacag tctagtatg 
      tacctcacc acggaagaa aaatgcaaa gcgttacaa 
      aactcagta tcgttgcgc gccctcttt gaggaaatt 
      tgtcgaacc acaacgggg attccctta ggaacgcat 
      agataccgt ttgaggact tacaccgat tgttttttg 
      ggttcggaa ttagtcgat tggttgatt tgtcaacaa 
      aaagccaat agtcgagtt caagcagct gcaataagt 
      caggcatta cttgaaggg ggttatatt gaatgtgtc 
      tcagatcct tgttctttt gttgatggt tacgccctc 
      tacaaacct ggcattttt gtaacacct gaagttcta 
      aatcataat tattttgaa gttccgaat caggaagaa 
      ccaatttgg gtacaacaa atcccacag gaatcaagc 
      accactgat tccgataac gagcaagtc tcagtcaaa 
      aaacgtggc cctttgacc tcttcatct tcgtacatg 
      ttggattta aatgttgaa gcgaacact gtgtattta 
      tcacgccct ctagcgtca agttatagc atccaatcg 
      ggtgattct ggtgatgtg agttgttgt gagacggat 
      attacaact gttcgaaca tctgaacaa agagaattt 
      gttcctgaa gctggttgg cacaatgcg tcaaatttg 
      cgcgaagaa aacggcgaa aaattggct tataatctc 
      ctaacggaa gcgtaccaa caacacgaa caaggttta 
      ttgaaacaa ttactagcg tctaatggt ctgttgtta 
      tcatgggct gacgtgata atcccactt tgtaacgaa 
      ataataaat gttatacga cctgacaaa aaccacgat 
      gctgaagat ttagatatt cgacattat ataaaattt 
      aagaaatta tcaggtggt tcacgaact gacactaaa 
      ctgataagt gggattgtt tttacgaaa aatgttgcc 
      cacaaagga atgttaacc gaaattgac aatccaaaa 
      atccttctt ttacaatgt tccattgtt taccaacga 
      acagaaggc cgtttaatg tcactagag ccggtcctg 
      atgcaagaa catgaatat ttacgtcac gtggctgct 
      cgtatcgtt gcccttcaa ccagatatc gttctcgtt 
      catcgaaac gtctcccga ttggctcaa gacctatta 
      cgtcagcac ggcatcacc ttggtgcac aacgtcaaa 
      caaagtgtt ttagaccaa ttggcacgt tgcacagag 
      gcggacttg gtcacggct gtagatgcc catattggg 
      cgcccccgt cttggcact tgtaaaaaa ttctatttg 
      aaaacgtac aatgttgat aaaggcgga gctaaaacg 
      ttgatgttt ttcgaaggt ttgcctatg gtgcattta 
      ggcgggact gtgctttta aggggcggg tctagacag 
      gagctcacc cgattggag aaagttgtt tcgttttgt 
      ttatttgcg agttataat tggcggtta gagaagtcg 
      tttttgatg gatgagttt gcgcagccg cctaatcca 
      aactgtgaa tttttggag gatagtagt aaggagagt 
      tcgccacgt ttgcaaact gttgtgaag tatgacagt 
      ggagggaag aagatgaat agtgagatg gttgaggat 
      tttacagat ccattacag tcatcattt gttgaaagt 
      atgaccgaa ggaaaggaa atcttagcg gttgcggaa 
      ttgccattt tcgaataat tttcgcaaa agtttagat 
      gattcgatt ttgtgtatt tcaccgtat ttgatgttt 
      tcggtgccg tatttggag acggaagtt ggaaaaaaa 
      tgtaaattg aggaagttt tttccgacg gaaatttat 
      ttcagtgaa caattcaca aatcagaca aaggtcaaa 
      atcattaag gatatcggc gaggatgtg aaagagaac 
      gagcagaaa tctgtcaag cccttacat ccatttcta 
      cgtacaaaa atcacaaca agtgttgat aataatgag 
      attcagtca ttattggct catttccgc gcttgtggt 
      ggcagttac gaaaagcgt gaaatttta tgtaaaccc 
      ccggtacaa gtcgacgag gaaatttta caaaacaag 
      gacaagaac tattacaat aaacttgat gttcttgaa 
      atcaacaat caccaaaga ctttcagta ttattttgt 
      agttttagt cacgagtcg accaattca ccggcattt 
      tgcgtaaat ccttggatt gttaatatg aatttttac 
      ggctcaaat gacattcca ttgggttgt tttctcgag 
      cgttattgt ttccgttca acttataat tgtccgtca 
      aagccttgt ggtacccca atgtttaaa catatccga 
      cgtttcgtc cacaatgca ggctgcgtc agtatctct 
      ctgaataat ttcgataac gaattcacc gaagaaaat 
      atcgttatg tggagttgg tgcaccaag tgtcaaagt 
      gtatcacca gtggtcctc atgtcggct gatacttgg 
      tcgttttct tttgccaaa tatctcgag ttaaaattt 
      tacggtggt ttgtatagt cgaagaggg aatacaccg 
      tgtggtcac agtttgcat catgatcac taccagtat 
      ttcggctac aaaaattca gtggcgtct ttcaagtat 
      acacccatt caagtgtgg gacatttct ctacctcca 
      cctgtgatc tacatacag tatgatata gaaaaacag 
      caaaccgaa ttaattgac gaaatacgc acaatggcg 
      caaaaaggc cacgaaatt tactctctc attttagaa 
      aaactgtct tgtattcca gccgaactt gaaggtcta 
      ggcaattta aaacaatta ctactaaaa gaacaaacc 
      caattcaag caaaaagtc gaagaagtg caattaaaa 
      ctcacctca ccgacaatt gaaaataaa caatttgac 
      gaaacaccc gaaaaacta catatcgct tattggaaa 
      atctccgat tcacttatc cgtatcaaa cgcctgatt 
      gttgaaact gtagacaat tggaacttg cgattatcc 
      gaaagtgca cgtaaacga gacgacaaa aagaaagac 
      cgaacttcg tacagcgat ctcgaaagt ccgacaata 
      cccgaaaat aaaacacta tcagaaaca gatatttca 
      cacagtgag acttcaaat agtgctacc ataacgaaa 
      aaaatcaaa agtttggat caaagtgac gagagtgaa 
      ttgagcatg tcttcatca cccaaatgt caccataga 
      tcgcaatca gatggtact gtaatgtca cacaatgag 
      gaacaaaac gattgtaag aaagacagt gacaaaaaa 
      accgtcaag aatatactg tcgcaactt ttgccctca 
      tcaagtgcc ctcacccta attccgaat ccgttcaat 
      ccgcaagac cactacacg ctcccgacc ggtgtttcg 
      gttcctata atagtctac gagtccgaa ccaagttcg 
      ataatagct tacgccttg aactcgtac gattataaa 
      aaatcgttt gaagatttg acgaaaaaa tcaaacact 
      gaacaaact cctagtccc atagtaaag cgcaaaaac 
      ccaaatacg gataaaaac gacgaaaca tcgggcctt 
      ttaggtttt ctacgaaac aaaaacgat ctcaattcg 
      ccggtatct gcatcggaa agtccacaa aatattgaa 
      gtgactgaa aaatcgaaa aatttacat attgaagtg 
      cagtttcaa gacacacat tgtaatttc ttctgtcgg 
      acttacttc gctgaaaaa tttgcgagt ttgaggggt 
      ttagtgttg cctattggt gaagagggt tatgttagg 
      tcattggct aggtcggtc caatggaat gctagagga 
      ggaaagtct ggttcgaat tttgctaaa acagccgat 
      gatcgtttc gtgttgaaa gaaatgtcc aaatccgaa 
      gtccagttg tttttagaa tcagcaccg aattatttt 
      aattacatg caaaagtgt tacgggacg ggacaaccg 
      acactttta ggcaaaatt ataggaatc tatcagatt 
      attttcaag aataataat tcaaacgtt actttgaga 
      acaaatcta ctggtcatg gagaatttg ttttacaag 
      cgtacagtg tcgcagaaa ttcgatttg aagggttca 
      atgaggaac agactggtc gtgcccgat aatcaagaa 
      ggcgaaatt gttttgttg gacgaaaat cttctcaaa 
      atgacttgt gatgctccg ctttacatt ttaccacac 
      tcgaaggcc gttctcaca gctgctatc cagaatgac 
      accgagttt ttatcagca cagtcagtg atggactat 
      tcgttgttg gtgggttta gattctgaa aaccgggag 
      ttggttctt ggaataata gattacatt cgaacgttc 
      acgtgggat aagaagctc gaaaccatg gtgaaaaaa 
      agtgggatt ttgggtggg caagggaaa ttaccaaca 
      attgtctct ccagaagag taccagaaa aggttcatt 
      gaagccatg catcgatat tttttggaa gttcccgat 
      cattgggct gggttgggc aaaggcttg gaattttga
    mRNA sequence
    • >TC003226 RA
      cacttgttt cacacgaca ctgaaaaca catcacttt 
      gttgacgat tcggttatt tataacaca caatataaa 
      aacacaacc tgacacatc tacaacaaa actgacact 
      tgcaggacg gattcaagg gggacgcaa agccccact 
      ttcaaacaa aactaccaa caaattaat gcgtcctgt 
      ctttgtcta acacgcaca attagaaaa agaaaaagt 
      aggaaaatg gttggtggt gctggcaat gctagcggc 
      ctcttgtgg caattttgc ataacggcc ttgacgaaa 
      ccacaagga cacaataaa aatacaaca atttggcac 
      aaaaaccac acaaacact gattattat ttaatttct 
      tatcattat caacacttt tatcagctt tagccgaca 
      attcacaga ttttatgtt acaagtaac aaatgacac 
      ttgacagtt gtaatttat aggttgtaa atacaagtc 
      agctgactt taaattgta atatttatg aattaatcg 
      ttgttaaaa tgtgatcag tgttgtgtt gtggccaag 
      atgaacaag aatctacaa tcccccacg aaattaact 
      gaattcgcg cccattcaa acggaagaa aagccccaa 
      actgtgggt cagttcatt accaagatt ttcaaattt 
      aataaaggc ccaaattcg gaagaacaa aactcgggt 
      ttaaccccg aattcggat aatgtgtct caggagtct 
      ctaccgaca tgggcaatc gagtcagac tcgtctgca 
      acccctttt cacgtagat gtgaacgag gggcgaagc 
      cttcccaat gttttgaaa cgaatcagt aatttacta 
      gcattaaaa accacgaac ttgcaagac tattcagac 
      accgaactg aaacaatat tggatgccg gattctgtg 
      agcaaagaa tgctaccaa tgttgcgaa aaattcact 
      acgttcaga cgccgacat cactgtcgt gtctgtggc 
      caaattttt tgctcacag tgttgcaat caacaaatt 
      ccgggaaaa attttcgga tgtactggt gatttaagg 
      gtttgcaca tattgctgc aaagtggtg ttatcatat 
      ctacagtcg tcagatgtt ggggctgac ctaactgcc 
      gatttaaag ctcctccaa gagaattta caaagcaaa 
      ttcggcaac gatgttctc tccccaact gtgcaagac 
      aacatacaa aatttgaac aaggattca gtcgaagtt 
      gggaatatt ttacgccgg aaaatttcc gtggggtat 
      caggaggag aaattcgca tctggacag tctagtatg 
      tacctcacc acggaagaa aaatgcaaa gcgttacaa 
      aactcagta tcgttgcgc gccctcttt gaggaaatt 
      tgtcgaacc acaacgggg attccctta ggaacgcat 
      agataccgt ttgaggact tacaccgat tgttttttg 
      ggttcggaa ttagtcgat tggttgatt tgtcaacaa 
      aaagccaat agtcgagtt caagcagct gcaataagt 
      caggcatta cttgaaggg ggttatatt gaatgtgtc 
      tcagatcct tgttctttt gttgatggt tacgccctc 
      tacaaacct ggcattttt gtaacacct gaagttcta 
      aatcataat tattttgaa gttccgaat caggaagaa 
      ccaatttgg gtacaacaa atcccacag gaatcaagc 
      accactgat tccgataac gagcaagtc tcagtcaaa 
      aaacgtggc cctttgacc tcttcatct tcgtacatg 
      ttggattta aatgttgaa gcgaacact gtgtattta 
      tcacgccct ctagcgtca agttatagc atccaatcg 
      ggtgattct ggtgatgtg agttgttgt gagacggat 
      attacaact gttcgaaca tctgaacaa agagaattt 
      gttcctgaa gctggttgg cacaatgcg tcaaatttg 
      cgcgaagaa aacggcgaa aaattggct tataatctc 
      ctaacggaa gcgtaccaa caacacgaa caaggttta 
      ttgaaacaa ttactagcg tctaatggt ctgttgtta 
      tcatgggct gacgtgata atcccactt tgtaacgaa 
      ataataaat gttatacga cctgacaaa aaccacgat 
      gctgaagat ttagatatt cgacattat ataaaattt 
      aagaaatta tcaggtggt tcacgaact gacactaaa 
      ctgataagt gggattgtt tttacgaaa aatgttgcc 
      cacaaagga atgttaacc gaaattgac aatccaaaa 
      atccttctt ttacaatgt tccattgtt taccaacga 
      acagaaggc cgtttaatg tcactagag ccggtcctg 
      atgcaagaa catgaatat ttacgtcac gtggctgct 
      cgtatcgtt gcccttcaa ccagatatc gttctcgtt 
      catcgaaac gtctcccga ttggctcaa gacctatta 
      cgtcagcac ggcatcacc ttggtgcac aacgtcaaa 
      caaagtgtt ttagaccaa ttggcacgt tgcacagag 
      gcggacttg gtcacggct gtagatgcc catattggg 
      cgcccccgt cttggcact tgtaaaaaa ttctatttg 
      aaaacgtac aatgttgat aaaggcgga gctaaaacg 
      ttgatgttt ttcgaaggt ttgcctatg gtgcattta 
      ggcgggact gtgctttta aggggcggg tctagacag 
      gagctcacc cgattggag aaagttgtt tcgttttgt 
      ttatttgcg agttataat tggcggtta gagaagtcg 
      tttttgatg gatgagttt gcgcagccg cctaatcca 
      aactgtgaa tttttggag gatagtagt aaggagagt 
      tcgccacgt ttgcaaact gttgtgaag tatgacagt 
      ggagggaag aagatgaat agtgagatg gttgaggat 
      tttacagat ccattacag tcatcattt gttgaaagt 
      atgaccgaa ggaaaggaa atcttagcg gttgcggaa 
      ttgccattt tcgaataat tttcgcaaa agtttagat 
      gattcgatt ttgtgtatt tcaccgtat ttgatgttt 
      tcggtgccg tatttggag acggaagtt ggaaaaaaa 
      tgtaaattg aggaagttt tttccgacg gaaatttat 
      ttcagtgaa caattcaca aatcagaca aaggtcaaa 
      atcattaag gatatcggc gaggatgtg aaagagaac 
      gagcagaaa tctgtcaag cccttacat ccatttcta 
      cgtacaaaa atcacaaca agtgttgat aataatgag 
      attcagtca ttattggct catttccgc gcttgtggt 
      ggcagttac gaaaagcgt gaaatttta tgtaaaccc 
      ccggtacaa gtcgacgag gaaatttta caaaacaag 
      gacaagaac tattacaat aaacttgat gttcttgaa 
      atcaacaat caccaaaga ctttcagta ttattttgt 
      agttttagt cacgagtcg accaattca ccggcattt 
      tgcgtaaat ccttggatt gttaatatg aatttttac 
      ggctcaaat gacattcca ttgggttgt tttctcgag 
      cgttattgt ttccgttca acttataat tgtccgtca 
      aagccttgt ggtacccca atgtttaaa catatccga 
      cgtttcgtc cacaatgca ggctgcgtc agtatctct 
      ctgaataat ttcgataac gaattcacc gaagaaaat 
      atcgttatg tggagttgg tgcaccaag tgtcaaagt 
      gtatcacca gtggtcctc atgtcggct gatacttgg 
      tcgttttct tttgccaaa tatctcgag ttaaaattt 
      tacggtggt ttgtatagt cgaagaggg aatacaccg 
      tgtggtcac agtttgcat catgatcac taccagtat 
      ttcggctac aaaaattca gtggcgtct ttcaagtat 
      acacccatt caagtgtgg gacatttct ctacctcca 
      cctgtgatc tacatacag tatgatata gaaaaacag 
      caaaccgaa ttaattgac gaaatacgc acaatggcg 
      caaaaaggc cacgaaatt tactctctc attttagaa 
      aaactgtct tgtattcca gccgaactt gaaggtcta 
      ggcaattta aaacaatta ctactaaaa gaacaaacc 
      caattcaag caaaaagtc gaagaagtg caattaaaa 
      ctcacctca ccgacaatt gaaaataaa caatttgac 
      gaaacaccc gaaaaacta catatcgct tattggaaa 
      atctccgat tcacttatc cgtatcaaa cgcctgatt 
      gttgaaact gtagacaat tggaacttg cgattatcc 
      gaaagtgca cgtaaacga gacgacaaa aagaaagac 
      cgaacttcg tacagcgat ctcgaaagt ccgacaata 
      cccgaaaat aaaacacta tcagaaaca gatatttca 
      cacagtgag acttcaaat agtgctacc ataacgaaa 
      aaaatcaaa agtttggat caaagtgac gagagtgaa 
      ttgagcatg tcttcatca cccaaatgt caccataga 
      tcgcaatca gatggtact gtaatgtca cacaatgag 
      gaacaaaac gattgtaag aaagacagt gacaaaaaa 
      accgtcaag aatatactg tcgcaactt ttgccctca 
      tcaagtgcc ctcacccta attccgaat ccgttcaat 
      ccgcaagac cactacacg ctcccgacc ggtgtttcg 
      gttcctata atagtctac gagtccgaa ccaagttcg 
      ataatagct tacgccttg aactcgtac gattataaa 
      aaatcgttt gaagatttg acgaaaaaa tcaaacact 
      gaacaaact cctagtccc atagtaaag cgcaaaaac 
      ccaaatacg gataaaaac gacgaaaca tcgggcctt 
      ttaggtttt ctacgaaac aaaaacgat ctcaattcg 
      ccggtatct gcatcggaa agtccacaa aatattgaa 
      gtgactgaa aaatcgaaa aatttacat attgaagtg 
      cagtttcaa gacacacat tgtaatttc ttctgtcgg 
      acttacttc gctgaaaaa tttgcgagt ttgaggggt 
      ttagtgttg cctattggt gaagagggt tatgttagg 
      tcattggct aggtcggtc caatggaat gctagagga 
      ggaaagtct ggttcgaat tttgctaaa acagccgat 
      gatcgtttc gtgttgaaa gaaatgtcc aaatccgaa 
      gtccagttg tttttagaa tcagcaccg aattatttt 
      aattacatg caaaagtgt tacgggacg ggacaaccg 
      acactttta ggcaaaatt ataggaatc tatcagatt 
      attttcaag aataataat tcaaacgtt actttgaga 
      acaaatcta ctggtcatg gagaatttg ttttacaag 
      cgtacagtg tcgcagaaa ttcgatttg aagggttca 
      atgaggaac agactggtc gtgcccgat aatcaagaa 
      ggcgaaatt gttttgttg gacgaaaat cttctcaaa 
      atgacttgt gatgctccg ctttacatt ttaccacac 
      tcgaaggcc gttctcaca gctgctatc cagaatgac 
      accgagttt ttatcagca cagtcagtg atggactat 
      tcgttgttg gtgggttta gattctgaa aaccgggag 
      ttggttctt ggaataata gattacatt cgaacgttc 
      acgtgggat aagaagctc gaaaccatg gtgaaaaaa 
      agtgggatt ttgggtggg caagggaaa ttaccaaca 
      attgtctct ccagaagag taccagaaa aggttcatt 
      gaagccatg catcgatat tttttggaa gttcccgat 
      cattgggct gggttgggc aaaggcttg gaattttga 
      tgtgttttt tttgtatta tattttatg taacacgtt 
      ttaataaaa accaattcg aatg