Status
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Gene information
General information
TC032418
Former TC numbers

If one gene is splitted or genes are merged, the original TC numbers are listed. One listed former TC number indicates the previous gene has be splitted, serveral former TC number represents a merged gene.

TC002859,TC002858,TC002860
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_seqid16: 3445614..3482129
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
    TC032418
    iBeetle screen iB_00458
    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: 100.0% (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: 9 (9 dpi)
    Days post injection


    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: 20.0% (includes death as larva, prepupa, pupa)
    Lethalities 22 days after larval injection: 100.0% (includes death as larva, prepupa, pupa, adult)

    larva lethality early (L5/6) - Number of animals showing the phenotype: 1 (11 dpi)
    Days post injection


    larva lethality late (L7) - Number of animals showing the phenotype: 1 (11 dpi)
    Days post injection


    pupal molt delayed (still larva) - Number of animals showing the phenotype: 8 (11 dpi)
    Days post injection


    pupal molt not fulfilled (died as L7 or prepupa) - Number of animals showing the phenotype: 8 (19 dpi)
    Days post injection


    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC032418 RA
      MASNFKYGS FILLILATT LAVDKSLFK TCEHSSFCR 
      RLRNFKPGS IFYVLNLDT LHVSGDNKA VVTANPIKV 
      EFFKNDVLV AVINGKGLF KIEHLRTKH QGEREEDSG 
      AWEEDFKSH HDSKPRGPE AVGVDVTFP GALRIYGLP 
      ERADKLALR STGSGGLDP HRLYNLDVY EYEVDSTMA 
      IYGAVPVVY AHSDKNTVG VFWHNAAET WVDINNSKD 
      KNFVSSIIN LVSSHQYDN FVDAHFMSE SGVFDMFVL 
      MGPKPKDAV RQYASLTGV APLPQYFAL GYHQCRWNY 
      FDETDVVNV VNNFDLNEL PVDSIWLDI EYTDGKKYF 
      TWQPFKFAH PREMVSNLT STGRKLVVI IDPHIKRED 
      GYFLHEDCL ANDYYVKNK DGNVYEGSC WPGASSYPD 
      LFDPKVQEY YKGLYALDK FKGTTQDVH IWNDMNEPS 
      VFNGPEVTM PKDCKHYGG WEHRHVHNL YALLYTEIT 
      YAGLIKRTS DRRPFILTR GHFAGSQRT AAVWTGDNA 
      AEWSHLQAS FPMCLAEAL GGISFCGAD IGGYFNNPD 
      IELLQRWYQ AGIWLPFYR AHAHLETKR REPYVFNED 
      VRTRIRNAL RLRYAILPL FYTLFREHE MTGEPVIRP 
      LFYEYPTDT NVVDIENQL LLGDRILAR PVTESGVSS 
      VSVYLPGGP EQFWYDIED FKQYQGTGV VNIPVTLDN 
      IPAFYRGGT ITPRKDRPR RSSTLTHHD PYTLYVALD 
      NNKSASGLL LISCFLQVS VVIAIMALN FKYGFLILL 
      ILATTSAVD KNNFKSCGQ SSFCRRLRN FKPGTISYV 
      LNLDTLHVG DNSIEADLK NVEAEADFR FTLTSLAGN 
      IFRVFIDEV KPLHPRYQV QGALNGEPQ VAKLEVLER 
      TKDKVTVRS GDNKAVVTA NPIKVEFFK NDALVAVIN 
      EKGLFNMEH LRAKPAEGE GEGEQEVKE DPGAWEENF 
      KSHHDSKPR GPEAVGVDV TFPGAFRIY GLPEHADRL 
      ALRSTGPGG LDPYRLYNL DVFEYEVDS TMAIYGAVP 
      VVYAHSDKN TVGVFWHNA AETWVDISN SKDTNVVSS 
      IVNLVSGHQ PDNAVDAHF MSESGVFDM FVLMGPKPK 
      DAVRQYASL TGVAPLPQY FALAYHQCR WNYNDETDV 
      INVIDNFDL NELPVDVIW LDIEYTDGK KYFTWDPVK 
      FAHPSEMIS NLTSTGRKL VVIIDPHIK REGGYFLHE 
      DCLANDYYV KNKDGNVYE GWCWPGSSS YPDLLDPKV 
      QEYYKGLYA LDKFKGTTQ DVHIWNDMN EPSVFNGPE 
      VTMPKDCKH YGGWEHRHI HNIYGLLYT EITYAGLIK 
      RSSDRRPFI LTRSHFAGS QRTAAVWTG DNAAEWSHL 
      QASFPMCLS EALGGISFC GADIGGFFN NPDTELLQR 
      WYQTGIWLP FYRAHAHLD TRRREPYLF NEDVRTRIR 
      NALRLRYAI LPLFYTLFR EHETTGEPV IRPLFYEYP 
      TDTNVIDID DQLLVGDRI LARPVTESG VSSVSVYLP 
      GGPEQFWYD IEDFKQYHG TGVVNIPVT LDKIPAFYR 
      GGTITPRKD RPRRSSTLT HHDPYTLYV ALDNNKSAS 
      GTLYVDDGE SFGYRNKKY LYLNFEFKD NTLTSSLID 
      KTDFPTDAW IERVVILGP PEKITKAIL TSNSLGTVN 
      LETSYDGEN RSLVVRKPG VNVREPFTL KLH
    CDS sequence
    • >TC032418 RA
      atggcgtca aatttcaag tacggttcc tttatttta 
      ttaatactc gccacaact ttagcagtt gataagagc 
      ctttttaag acatgcgaa catagtagt ttttgcagg 
      cgcttgaga aattttaaa cctggatcg attttttac 
      gttttaaat ttagacact ctacacgtc tctggcgat 
      aacaaagcc gtcgttaca gcaaatccc attaaggtc 
      gagtttttc aaaaatgac gtccttgtg gctgttatc 
      aatgggaag gggttgttc aaaatagaa catctcagg 
      actaaacac cagggtgaa agagaagag gactccggt 
      gcttgggaa gaagatttc aagtcgcat catgattcc 
      aaacctagg gggcctgaa gcagtgggt gttgatgtc 
      acttttcct ggagctttg aggatctat ggactgcct 
      gaacgtgca gataagctg gctttgagg agcacgggt 
      tctgggggg cttgacccc catcgattg tataatttg 
      gatgtttat gagtatgaa gttgactcc actatggcc 
      atttatggg gctgtaccg gtggtctat gctcattca 
      gataaaaac acggtgggc gttttttgg cacaacgcc 
      gccgaaacc tgggttgac atcaacaac agcaaagac 
      aaaaatttt gtttcttcc atcataaac ctagtctca 
      agccaccaa tatgacaat ttcgttgat gctcatttc 
      atgagtgag agtggcgtt ttcgatatg tttgtttta 
      atgggacct aagccgaaa gatgccgta cgacaatac 
      gcatctttg accggtgtt gccccctta ccacaatat 
      tttgcgcta ggctatcac caatgccgc tggaactac 
      tttgacgaa actgacgtc gtcaacgtt gttaataat 
      tttgatctc aatgaatta cctgttgat tctatttgg 
      ctagatatt gagtacacg gatgggaaa aagtatttc 
      acgtggcag ccgttcaaa tttgcacat ccgcgtgaa 
      atggtttcg aatttgaca tcaactggc cgaaagttg 
      gttgttatc attgatccg catattaag cgagaggat 
      ggttatttc ctccatgag gattgtcta gctaatgac 
      tattacgtt aaaaataaa gacggcaat gtttacgaa 
      ggttcctgc tggcctggt gctagcagt tacccggat 
      ttgttcgat cctaaagtc caggagtac tacaaaggt 
      ttgtacgct ttggataag ttcaaaggg acgactcaa 
      gacgtccac atttggaac gatatgaac gaaccgtcg 
      gtttttaac ggaccggag gtcacaatg cctaaagat 
      tgtaaacat tacggaggc tgggaacac agacatgtg 
      cacaacctt tacgctttg ctttatact gaaatcact 
      tatgccggt ttgattaaa cggacaagt gaccgcagg 
      ccgtttatt ttaacaagg ggtcacttt gccggatct 
      cagagaact gcagctgtt tggactgga gataacgcg 
      gcagaatgg tcgcatctc caggccagt tttccaatg 
      tgtttggct gaagctttg ggagggatt agtttttgt 
      ggggctgat attggaggg tactttaat aatcctgat 
      atcgaatta ttgcaaagg tggtaccaa gccggtatt 
      tggttacca ttttaccgt gcccatgcc catctagag 
      acaaaacgt cgcgaacct tacgttttc aacgaagat 
      gttcgtact agaatccga aacgctttg aggctacgt 
      tatgccatt ttgcccctg ttttacacc ttgtttagg 
      gaacatgaa atgacaggg gaacctgtg ataaggcca 
      ctattttac gagtatcct actgatact aatgttgtc 
      gatattgaa aaccaacta cttctcggt gaccgaatt 
      ttggcacga cctgtgacc gagtccggc gtgtcttcg 
      gtcagtgtt tacctgcct ggaggtcct gaacaattt 
      tggtacgat attgaagat ttcaaacaa tatcaggga 
      actggcgtt gttaacata ccggtgact ttagataat 
      attcctgcg ttttatagg ggtggcact atcacccca 
      cgtaaggat aggcctagg aggtcgtcc actttaaca 
      catcatgat ccttacaca ctttacgtt gcattggac 
      aacaacaaa tctgcgtcc ggtttattg ctaatcagc 
      tgtttttta caagtgtca gtcgtcatc gccatcatg 
      gcgttaaat tttaagtac ggtttcttg attttatta 
      atactcgca acaacatca gcggtcgac aaaaacaat 
      tttaaatca tgcggacaa agtagtttt tgcaggcgc 
      ttgagaaat ttcaagcct ggaacgatt tcctacgtt 
      ttaaattta gacacttta cacgttggt gataactca 
      atagaggcc gatttgaaa aatgtggaa gccgaagct 
      gattttaga ttcactttg acttctctt gctggaaat 
      atattcagg gtgtttatt gatgaagtc aagcctttg 
      catccccgg tatcaagta cagggggca cttaatggc 
      gagccacaa gtggccaaa ttggaagtt ttagagcgc 
      acaaaagac aaagtaacg gtacgttca ggcgataac 
      aaagcagtc gtcacagca aatcctatt aaagttgag 
      tttttcaaa aatgacgcc cttgtggct gttatcaat 
      gagaagggg ttattcaac atggaacat ctcagggct 
      aagcctgcc gaaggtgaa ggggaaggg gaacaagag 
      gtgaaggaa gaccccggc gcttgggaa gaaaatttc 
      aagtctcat catgactcc aaacctaga ggacctgaa 
      gcagtgggt gttgatgtc acttttcca ggagccttc 
      aggatttat gggctgcct gaacatgca gataggttg 
      gctttgagg agcacgggg cctgggggg cttgaccct 
      taccgcttg tacaatttg gacgttttt gagtatgaa 
      gttgactcc actatggcc atctatggt gctgtcccg 
      gtggtctat gctcattca gacaaaaac acagtgggc 
      gttttctgg cacaacgcc gccgaaacc tgggtagac 
      atcagcaac agtaaagac acaaacgtt gtttcctcc 
      atcgtaaac ctggtctca ggccaccag cctgacaac 
      gccgttgac gctcatttc atgagcgag agtggcgtt 
      ttcgatatg tttgtttta atgggacct aagccgaaa 
      gatgccgta cgacaatac gcctctttg accggtgtt 
      gccccctta ccacaatac tttgcacta gcctaccac 
      cagtgccgt tggaactac aacgacgaa actgacgtg 
      attaacgtc atagataat ttcgatctc aatgagtta 
      cctgtcgat gttatctgg ctagatatt gaatacacc 
      gatgggaaa aaatatttc acgtgggat ccggttaag 
      tttgcacat ccgagcgaa atgatttcg aatttgaca 
      tcaactggc cgaaagttg gtcgttatt attgatccg 
      catattaag cgagagggt ggttatttc ctccatgag 
      gactgtcta gccaatgac tattacgtt aaaaataaa 
      gacggcaat gtttacgaa ggttggtgc tggcctggg 
      tctagtagt tacccggat ttgctcgat cctaaagtc 
      caggagtac tacaaaggt ttgtacgct ctggataag 
      ttcaaaggg acgactcaa gacgtccac atttggaac 
      gatatgaac gaaccgtcg gtttttaac ggaccggag 
      gtcacaatg ccgaaagat tgcaaacat tacggaggc 
      tgggaacat agacacatt cacaacatc tacgggttg 
      ctttatact gaaatcact tatgccggt ttgattaag 
      cggtcaagt gaccgcagg ccgtttatt ttaacaagg 
      agtcacttt gccggatct cagagaact gcagctgtt 
      tggactgga gataacgcg gcagaatgg tcgcatctc 
      caggccagt tttccaatg tgtttatcc gaggctttg 
      ggagggatt agtttttgt ggggctgat attggaggc 
      ttctttaat aatcctgat accgagttg ttgcaaagg 
      tggtaccag accgggatc tggttacca ttttaccgt 
      gcccatgcc catctggac acaagacgt cgcgaacct 
      taccttttc aatgaagat gttcgtact agaatccga 
      aacgctttg agactacgt tatgccatt ttgcccctg 
      ttttacacc ttgttcagg gaacatgaa acgacaggg 
      gaacctgtg ataaggcca ctattttac gagtatcct 
      actgatact aatgttatc gatatcgac gaccaactt 
      ctagtcggt gaccggatt ttggcccga cctgtgacg 
      gagtccggc gtgtcttcc gtcagtgtt tacctgcct 
      ggaggtcct gaacaattt tggtacgat attgaagat 
      ttcaaacaa tatcacgga actggcgtt gttaacata 
      ccggtgact ttagataag attcctgcg ttttatagg 
      ggtggcact atcacccca cgtaaagat aggccgagg 
      aggtcgtcc actttaaca catcatgat ccttacaca 
      ctttacgtt gcattggac aacaataaa tctgcgtcc 
      ggtacgttg tatgtagat gacggtgaa agttttggt 
      tatcgtaat aagaaatac ttgtatttg aacttcgag 
      tttaaggac aatactctg accagcagc ttgattgat 
      aaaactgat ttccctact gatgcctgg atcgagcga 
      gtggtcatt ttaggaccg cctgagaaa atcaccaag 
      gctatttta acgagcaat agcttaggt actgttaat 
      ttggaaact tcttacgac ggtgaaaat cggtccttg 
      gtcgttcgt aagcctggt gttaatgtt agagagccg 
      tttacgttg aaacttcat taa
    mRNA sequence
    • >TC032418 RA
      gttaaatat taatccatt aatgagaaa aaattatta 
      ggtttattg ctcttacga cgagttaat catcatagc 
      caccatggc gtcaaattt caagtacgg ttcctttat 
      tttattaat actcgccac aactttagc agttgataa 
      gagcctttt taagacatg cgaacatag tagtttttg 
      caggcgctt gagaaattt taaacctgg atcgatttt 
      ttacgtttt aaatttaga cactctaca cgtctctgg 
      cgataacaa agccgtcgt tacagcaaa tcccattaa 
      ggtcgagtt tttcaaaaa tgacgtcct tgtggctgt 
      tatcaatgg gaaggggtt gttcaaaat agaacatct 
      caggactaa acaccaggg tgaaagaga agaggactc 
      cggtgcttg ggaagaaga tttcaagtc gcatcatga 
      ttccaaacc tagggggcc tgaagcagt gggtgttga 
      tgtcacttt tcctggagc tttgaggat ctatggact 
      gcctgaacg tgcagataa gctggcttt gaggagcac 
      gggttctgg ggggcttga cccccatcg attgtataa 
      tttggatgt ttatgagta tgaagttga ctccactat 
      ggccattta tggggctgt accggtggt ctatgctca 
      ttcagataa aaacacggt gggcgtttt ttggcacaa 
      cgccgccga aacctgggt tgacatcaa caacagcaa 
      agacaaaaa ttttgtttc ttccatcat aaacctagt 
      ctcaagcca ccaatatga caatttcgt tgatgctca 
      tttcatgag tgagagtgg cgttttcga tatgtttgt 
      tttaatggg acctaagcc gaaagatgc cgtacgaca 
      atacgcatc tttgaccgg tgttgcccc cttaccaca 
      atattttgc gctaggcta tcaccaatg ccgctggaa 
      ctactttga cgaaactga cgtcgtcaa cgttgttaa 
      taattttga tctcaatga attacctgt tgattctat 
      ttggctaga tattgagta cacggatgg gaaaaagta 
      tttcacgtg gcagccgtt caaatttgc acatccgcg 
      tgaaatggt ttcgaattt gacatcaac tggccgaaa 
      gttggttgt tatcattga tccgcatat taagcgaga 
      ggatggtta tttcctcca tgaggattg tctagctaa 
      tgactatta cgttaaaaa taaagacgg caatgttta 
      cgaaggttc ctgctggcc tggtgctag cagttaccc 
      ggatttgtt cgatcctaa agtccagga gtactacaa 
      aggtttgta cgctttgga taagttcaa agggacgac 
      tcaagacgt ccacatttg gaacgatat gaacgaacc 
      gtcggtttt taacggacc ggaggtcac aatgcctaa 
      agattgtaa acattacgg aggctggga acacagaca 
      tgtgcacaa cctttacgc tttgcttta tactgaaat 
      cacttatgc cggtttgat taaacggac aagtgaccg 
      caggccgtt tattttaac aaggggtca ctttgccgg 
      atctcagag aactgcagc tgtttggac tggagataa 
      cgcggcaga atggtcgca tctccaggc cagttttcc 
      aatgtgttt ggctgaagc tttgggagg gattagttt 
      ttgtggggc tgatattgg agggtactt taataatcc 
      tgatatcga attattgca aaggtggta ccaagccgg 
      tatttggtt accatttta ccgtgccca tgcccatct 
      agagacaaa acgtcgcga accttacgt tttcaacga 
      agatgttcg tactagaat ccgaaacgc tttgaggct 
      acgttatgc cattttgcc cctgtttta caccttgtt 
      tagggaaca tgaaatgac aggggaacc tgtgataag 
      gccactatt ttacgagta tcctactga tactaatgt 
      tgtcgatat tgaaaacca actacttct cggtgaccg 
      aattttggc acgacctgt gaccgagtc cggcgtgtc 
      ttcggtcag tgtttacct gcctggagg tcctgaaca 
      attttggta cgatattga agatttcaa acaatatca 
      gggaactgg cgttgttaa cataccggt gactttaga 
      taatattcc tgcgtttta taggggtgg cactatcac 
      cccacgtaa ggataggcc taggaggtc gtccacttt 
      aacacatca tgatcctta cacacttta cgttgcatt 
      ggacaacaa caaatctgc gtccggttt attgctaat 
      cagctgttt tttacaagt gtcagtcgt catcgccat 
      catggcgtt aaattttaa gtacggttt cttgatttt 
      attaatact cgcaacaac atcagcggt cgacaaaaa 
      caattttaa atcatgcgg acaaagtag tttttgcag 
      gcgcttgag aaatttcaa gcctggaac gatttccta 
      cgttttaaa tttagacac tttacacgt tggtgataa 
      ctcaataga ggccgattt gaaaaatgt ggaagccga 
      agctgattt tagattcac tttgacttc tcttgctgg 
      aaatatatt cagggtgtt tattgatga agtcaagcc 
      tttgcatcc ccggtatca agtacaggg ggcacttaa 
      tggcgagcc acaagtggc caaattgga agttttaga 
      gcgcacaaa agacaaagt aacggtacg ttcaggcga 
      taacaaagc agtcgtcac agcaaatcc tattaaagt 
      tgagttttt caaaaatga cgcccttgt ggctgttat 
      caatgagaa ggggttatt caacatgga acatctcag 
      ggctaagcc tgccgaagg tgaagggga aggggaaca 
      agaggtgaa ggaagaccc cggcgcttg ggaagaaaa 
      tttcaagtc tcatcatga ctccaaacc tagaggacc 
      tgaagcagt gggtgttga tgtcacttt tccaggagc 
      cttcaggat ttatgggct gcctgaaca tgcagatag 
      gttggcttt gaggagcac ggggcctgg ggggcttga 
      cccttaccg cttgtacaa tttggacgt ttttgagta 
      tgaagttga ctccactat ggccatcta tggtgctgt 
      cccggtggt ctatgctca ttcagacaa aaacacagt 
      gggcgtttt ctggcacaa cgccgccga aacctgggt 
      agacatcag caacagtaa agacacaaa cgttgtttc 
      ctccatcgt aaacctggt ctcaggcca ccagcctga 
      caacgccgt tgacgctca tttcatgag cgagagtgg 
      cgttttcga tatgtttgt tttaatggg acctaagcc 
      gaaagatgc cgtacgaca atacgcctc tttgaccgg 
      tgttgcccc cttaccaca atactttgc actagccta 
      ccaccagtg ccgttggaa ctacaacga cgaaactga 
      cgtgattaa cgtcataga taatttcga tctcaatga 
      gttacctgt cgatgttat ctggctaga tattgaata 
      caccgatgg gaaaaaata tttcacgtg ggatccggt 
      taagtttgc acatccgag cgaaatgat ttcgaattt 
      gacatcaac tggccgaaa gttggtcgt tattattga 
      tccgcatat taagcgaga gggtggtta tttcctcca 
      tgaggactg tctagccaa tgactatta cgttaaaaa 
      taaagacgg caatgttta cgaaggttg gtgctggcc 
      tgggtctag tagttaccc ggatttgct cgatcctaa 
      agtccagga gtactacaa aggtttgta cgctctgga 
      taagttcaa agggacgac tcaagacgt ccacatttg 
      gaacgatat gaacgaacc gtcggtttt taacggacc 
      ggaggtcac aatgccgaa agattgcaa acattacgg 
      aggctggga acatagaca cattcacaa catctacgg 
      gttgcttta tactgaaat cacttatgc cggtttgat 
      taagcggtc aagtgaccg caggccgtt tattttaac 
      aaggagtca ctttgccgg atctcagag aactgcagc 
      tgtttggac tggagataa cgcggcaga atggtcgca 
      tctccaggc cagttttcc aatgtgttt atccgaggc 
      tttgggagg gattagttt ttgtggggc tgatattgg 
      aggcttctt taataatcc tgataccga gttgttgca 
      aaggtggta ccagaccgg gatctggtt accatttta 
      ccgtgccca tgcccatct ggacacaag acgtcgcga 
      accttacct tttcaatga agatgttcg tactagaat 
      ccgaaacgc tttgagact acgttatgc cattttgcc 
      cctgtttta caccttgtt cagggaaca tgaaacgac 
      aggggaacc tgtgataag gccactatt ttacgagta 
      tcctactga tactaatgt tatcgatat cgacgacca 
      acttctagt cggtgaccg gattttggc ccgacctgt 
      gacggagtc cggcgtgtc ttccgtcag tgtttacct 
      gcctggagg tcctgaaca attttggta cgatattga 
      agatttcaa acaatatca cggaactgg cgttgttaa 
      cataccggt gactttaga taagattcc tgcgtttta 
      taggggtgg cactatcac cccacgtaa agataggcc 
      gaggaggtc gtccacttt aacacatca tgatcctta 
      cacacttta cgttgcatt ggacaacaa taaatctgc 
      gtccggtac gttgtatgt agatgacgg tgaaagttt 
      tggttatcg taataagaa atacttgta tttgaactt 
      cgagtttaa ggacaatac tctgaccag cagcttgat 
      tgataaaac tgatttccc tactgatgc ctggatcga 
      gcgagtggt cattttagg accgcctga gaaaatcac 
      caaggctat tttaacgag caatagctt aggtactgt 
      taatttgga aacttctta cgacggtga aaatcggtc 
      cttggtcgt tcgtaagcc tggtgttaa tgttagaga 
      gccgtttac gttgaaact tcattaaaa tttgttggt 
      ttagcattt ttcttaatt tttatttaa tacgtagaa 
      tgtttttta tgtgatact tccttccat cgttgtgct 
      acattttgt gtcaataaa gaaatttta cagag