Status
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Gene information
General information
TC031264
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.

TC002219,TC002218,TC002216
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_007423.3: 1036914..1089520
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
    TC031264
    iBeetle screen iB_03324

    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: 100.0% (includes death as pupa, adult)

    eclosion not fulfilled (with pupal and adult features) - Number of animals showing the phenotype: 2 (3 dpi)
    Days post injection


    adult lethality - Number of animals showing the phenotype: 8 (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: % (includes death as larva, prepupa, pupa)
    Lethalities 22 days after larval injection: % (includes death as larva, prepupa, pupa, adult)

    iBeetle screen iB_03323

    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: 20.0% (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: 1 (9 dpi)
    Days post injection


    Analysis of larval stage 1 cuticle
    number of eggs/embryos/larvae on slide: < 20
    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
    • >TC031264 RA
      MSYIEDANE TDSNNTSLD AFYFFETEQ FTLLWVLFV 
      IIVAGNVGV LYTLLFGRS RKSRMNYFI THLALADLS 
      VGLINVLTD IIWKTTVAW YAGNVACKV IRFLQVVVT 
      YASTYVLVA LSIDRYDAI RHPMKFSGS WKRARYLIL 
      AAWFFSALY SLPILIFYE EKIVKGKLQ CWIEFHPQW 
      KWQVYMTLV SLSLFIIPA SIIATCYAI IIITIWSKN 
      AKGFIKNTK AVNGSDNSR RASSRGLIP RAKVKTIKI 
      TFVIVSVFI LCWSPYIIF DLLQVFGQI PGTQTNIAI 
      ATFIQSLAP LNSAANPII YCLFSTHFC RTLGSLPPF 
      KWLFKKKKR RNRESTTNT QSSSLSEFL TNTHKRKIE 
      NTSTTFNAH TTCAVLFYL TSVFNSVLN LNEDRSYWL 
      DVGKNDINR ALNLEKIEN PSKNVILFV GDGMGLTTT 
      TASRIYSKS ESGFLSFEQ FPHIATIKT YNADKLVPD 
      SSSTATALF SGVKNNLKT SGVDSSVKL NDCEASLRP 
      EARIDGLMK WAQEAGKST GFVTTTRVT HATPSALYA 
      HTPNRNWEC ESKIPEKSK QCKDIARQL VEDEPGRNI 
      NVVMGGGRQ CLQSGVEDK PGDLVDRWA CYSLDGRDL 
      IRDWEMDKK KRNKTYATV LNSEELKAV DTSNEFVLG 
      IFANGHMSM DYARDKSPT GMPSLTNMT QKAIEILSQ 
      NPEGYILVV EGGLIDFAH HRGNARKAL DETVSLSDA 
      VQIAVEKTD PRETLIIVT SDHSHSLVF TGYPSRNKS 
      VLDIAQKSK IDHIPYTSL LYGTGGPNN YQYSVINNT 
      ISRQDPSIN NTSDFEYSQ QAVVLTDEV THGGSDVLI 
      YAKGPMSHL FHSVHEQTY VAYVISYAA KIGPFKNGS 
      SSFIYTKQI LKCSKIALF TRLYSSRNL LKLKERGMY 
      QDIFPDTAT NTVTDLLNA APQRVYAGF DPTADSLHI 
      GNLLVLINL IHWQRGGHQ PIALIGGAT AKIGDPSGK 
      TSEREKLAS AFVDENVIG IRNNIETLF YNHEKYLWK 
      KKGKLAPLI IVNNESWYS KMSPIDLIG GAGRYLRMG 
      TLLSRTSVQ TRLQSPAGM SFTEFSYQL FQAYDWLHL 
      FQKYNCSFQ IGGNDQMGN IMSGHELIS KTCDKQVYA 
      LTVPIITTE MGDKFGKSA GNAMWLSAE KTSPFTLYQ 
      FFVRQVDTE VEKMLKFFT FEPVGSIKD LMRRHTEKP 
      ELRLPQKYL AEQVVLLVH GEKGLEAAH KATKVLYER 
      SITALAEMN PLEIKSLFE GATVVEILP EPGQSVLDL 
      AMSAGCFLT KEDALRIIS AGGFYINHQ KTNNPNEVL 
      NLNVHRLSN NVTLLRVGK KNYYVVKWL S
    CDS sequence
    • >TC031264 RA
      atgtcatac attgaagat gcaaacgag accgattcg 
      aacaacaca tcgttagac gctttttac tttttcgag 
      accgaacaa ttcacctta ctatgggtc ctattcgtg 
      ataatagtc gctggaaac gtaggtgtg ttgtacacg 
      cttctcttt ggccgatct agaaaaagt cgaatgaat 
      tatttcata acccaccta gccctagca gaccttagt 
      gtgggtttg attaatgtt ttgacagat ataatttgg 
      aaaacaaca gtggcatgg tatgccgga aacgttgcc 
      tgtaaagtg atacgtttt ctccaggta gtggtaaca 
      tatgcctcg acctatgtt ttggttgct ttaagtata 
      gaccgatat gacgcaatt cgtcacccc atgaaattt 
      tcaggcagt tggaaacgt gctagatat ctaatatta 
      gccgcttgg ttcttcagt gcgctgtat tcgttaccg 
      attttgatc ttctacgag gagaagatt gtgaaagga 
      aaacttcaa tgttggatt gaatttcat ccacaatgg 
      aaatggcaa gtctacatg acactagtt tcgctctca 
      ctctttatc attcctgcg tcaattatt gcgacgtgt 
      tacgccatc attattatc acaatttgg tcgaaaaat 
      gccaaaggt ttcatcaaa aataccaaa gctgttaat 
      gggtctgat aattcaaga agggcaagt tcgagaggt 
      ttaatacca agagccaaa gtcaagact ataaaaatc 
      acatttgtt atagtgagc gtttttatt ctatgttgg 
      agtccttat ataattttt gatttgtta caagtgttt 
      gggcaaata ccagggact cagaccaat atagctatc 
      gctactttt atccaaagt ttagcacca ctcaactct 
      gcagctaac cccattatt tactgcctc ttttctaca 
      cacttctgt cggacattg gggtcattg cctccgttc 
      aaatggttg ttcaagaaa aagaaaaga aggaatagg 
      gaatcaaca acaaacaca caaagtagt tcattatcc 
      gaattctta accaacact cacaaacga aaaatcgaa 
      aatacatca acaactttt aacgcccat acaacatgt 
      gcagtatta ttctatttg acaagcgtg ttcaacagt 
      gtacttaat ttaaatgaa gacagatct tactggttg 
      gatgtagga aaaaacgat atcaacaga gctcttaac 
      ttagaaaaa attgaaaac ccttcaaaa aacgttata 
      ttgtttgtt ggtgatgga atgggctta acaactact 
      acagcatca agaatttac tccaagtct gaaagtggg 
      tttttgtct tttgagcaa tttcctcat atagcaacg 
      ataaagact tacaacgct gacaaattg gttcccgat 
      tcaagcagt acagcgacg gctcttttt tctggtgtg 
      aaaaacaat ttgaaaacg tcaggcgtt gattctagt 
      gtgaaatta aacgactgt gaagcctca cttagacca 
      gaagcgcgg attgacggc ttaatgaaa tgggcacaa 
      gaggctgga aaatcaact ggattcgta acaacgaca 
      cgcgtcaca catgccacc cccagtgca ctttacgca 
      cacaccccc aacagaaac tgggagtgt gaaagcaaa 
      attcccgaa aaatcgaaa caatgcaaa gacattgcg 
      agacaattg gttgaggat gagccgggg agaaatatt 
      aatgtagtg atgggtgga ggaagacag tgtttacaa 
      tcgggggtt gaagataaa ccaggtgat cttgttgat 
      agatgggca tgttatagt ttagatggg cgagatttg 
      ataagagac tgggagatg gacaaaaag aaaagaaat 
      aaaacttat gccactgtg ttgaatagt gaagaattg 
      aaggcagta gacaccagt aatgaattt gttttagga 
      atttttgca aatggacac atgtcaatg gattacgcc 
      cgagataaa tcaccaact ggaatgcct tcactcacg 
      aacatgaca caaaaagca attgaaatt ctaagccaa 
      aacccagaa ggatatatt ttggtagta gaaggtggt 
      cttatcgat tttgcccat cacagaggc aacgctaga 
      aaagctcta gacgagact gtctcactt agtgatgca 
      gttcagata gctgtcgaa aaaacggat ccacgagag 
      acattaatt attgtaact agtgatcac tcgcattct 
      ttggtattt actggatat ccttcaagg aacaaaagt 
      gtattagac attgcacaa aagtcaaaa attgaccac 
      ataccctac acttcgtta ctatatgga actggaggt 
      ccaaacaat tatcaatac tccgttatt aacaatacc 
      atttcaaga caagatcct tcaattaac aatacttca 
      gattttgaa tattcacaa caagctgta gttctaacg 
      gatgaagtg actcatggt ggaagcgat gttttaatc 
      tatgctaaa ggtccgatg tctcattta ttccacagt 
      gtacatgaa cagacgtat gttgcttac gtaattagt 
      tacgccgca aaaattgga ccatttaaa aatggaagt 
      tcatccttt atctacacg aaacaaatt ttaaaatgc 
      agcaagatt gcgcttttc acgcgactc tattccagc 
      cgaaacctt ctaaaattg aaagagcgc ggaatgtac 
      caagacatt ttcccagat acagcaaca aacacggtt 
      acggattta ttaaatgca gccccacag cgtgtttat 
      gcaggcttt gaccccact gcagacagc cttcatatc 
      ggaaattta cttgttctt attaattta atacactgg 
      caaaggggt ggccaccaa cccatagca ctgattgga 
      ggtgctacc gctaaaatt ggcgatccg agtggtaaa 
      acatcagaa agagaaaaa ttagcaagt gcttttgtt 
      gatgagaat gtgattgga attagaaac aacattgaa 
      acgcttttt tataatcat gaaaaatac ctatggaag 
      aagaaggga aaactggct cctctaata atcgtgaat 
      aacgaatct tggtatagt aaaatgtca ccaatcgat 
      ttgattggt ggtgctggt cgttatttg cgaatggga 
      acattatta agcagaaca agtgttcaa acacgcctt 
      cagtctccc gctggtatg agttttacc gaatttagt 
      taccaatta tttcaagca tatgattgg ctgcattta 
      tttcaaaaa tacaattgc tcttttcaa attggtgga 
      aatgatcaa atgggcaat attatgtcg gggcatgaa 
      ctcatcagc aaaacgtgc gacaaacaa gtttatgct 
      cttactgtt ccgataatt acaaccgaa atgggcgac 
      aaattcggc aaatctgca ggaaacgcc atgtggcta 
      tctgctgaa aaaacctca cctttcact ctttaccag 
      tttttcgtc cgtcaagtt gacaccgaa gttgaaaaa 
      atgttaaag ttcttcaca tttgaacca gtcggaagc 
      attaaagat ctgatgaga cgccacacc gaaaaacct 
      gaattgcga ctacctcaa aaatatctt gctgagcaa 
      gtggtgctc ttagtgcat ggggaaaag ggtttagag 
      gctgcgcat aaagcgaca aaagtttta tacgaacga 
      agcattaca gcactcgct gaaatgaat cctttggaa 
      ataaagagt ttatttgaa ggagccaca gttgtggaa 
      attttacca gagcctgga caaagtgtg ctagatttg 
      gcgatgagc gctggatgt tttcttacc aaagaagat 
      gcactgaga atcatttca gccggcggt ttttacatc 
      aaccatcaa aaaacaaac aatcctaat gaagtttta 
      aacttaaat gtacatagg ttaagcaat aatgtaacg 
      ttacttcga gtaggcaaa aagaactat tatgtagta 
      aaatggctt tcataa
    mRNA sequence
    • >TC031264 RA
      tctccgaat ttttgtgct agaaacggg caacatgta 
      acaatttct tgagacatg tcatacatt gaagatgca 
      aacgagacc gattcgaac aacacatcg ttagacgct 
      ttttacttt ttcgagacc gaacaattc accttacta 
      tgggtccta ttcgtgata atagtcgct ggaaacgta 
      ggtgtgttg tacacgctt ctctttggc cgatctaga 
      aaaagtcga atgaattat ttcataacc cacctagcc 
      ctagcagac cttagtgtg ggtttgatt aatgttttg 
      acagatata atttggaaa acaacagtg gcatggtat 
      gccggaaac gttgcctgt aaagtgata cgttttctc 
      caggtagtg gtaacatat gcctcgacc tatgttttg 
      gttgcttta agtatagac cgatatgac gcaattcgt 
      caccccatg aaattttca ggcagttgg aaacgtgct 
      agatatcta atattagcc gcttggttc ttcagtgcg 
      ctgtattcg ttaccgatt ttgatcttc tacgaggag 
      aagattgtg aaaggaaaa cttcaatgt tggattgaa 
      tttcatcca caatggaaa tggcaagtc tacatgaca 
      ctagtttcg ctctcactc tttatcatt cctgcgtca 
      attattgcg acgtgttac gccatcatt attatcaca 
      atttggtcg aaaaatgcc aaaggtttc atcaaaaat 
      accaaagct gttaatggg tctgataat tcaagaagg 
      gcaagttcg agaggttta ataccaaga gccaaagtc 
      aagactata aaaatcaca tttgttata gtgagcgtt 
      tttattcta tgttggagt ccttatata atttttgat 
      ttgttacaa gtgtttggg caaatacca gggactcag 
      accaatata gctatcgct acttttatc caaagttta 
      gcaccactc aactctgca gctaacccc attatttac 
      tgcctcttt tctacacac ttctgtcgg acattgggg 
      tcattgcct ccgttcaaa tggttgttc aagaaaaag 
      aaaagaagg aatagggaa tcaacaaca aacacacaa 
      agtagttca ttatccgaa ttcttaacc aacactcac 
      aaacgaaaa atcgaaaat acatcaaca acttttaac 
      gcccataca acatgtgca gtattattc tatttgaca 
      agcgtgttc aacagtgta cttaattta aatgaagac 
      agatcttac tggttggat gtaggaaaa aacgatatc 
      aacagagct cttaactta gaaaaaatt gaaaaccct 
      tcaaaaaac gttatattg tttgttggt gatggaatg 
      ggcttaaca actactaca gcatcaaga atttactcc 
      aagtctgaa agtgggttt ttgtctttt gagcaattt 
      cctcatata gcaacgata aagacttac aacgctgac 
      aaattggtt cccgattca agcagtaca gcgacggct 
      cttttttct ggtgtgaaa aacaatttg aaaacgtca 
      ggcgttgat tctagtgtg aaattaaac gactgtgaa 
      gcctcactt agaccagaa gcgcggatt gacggctta 
      atgaaatgg gcacaagag gctggaaaa tcaactgga 
      ttcgtaaca acgacacgc gtcacacat gccaccccc 
      agtgcactt tacgcacac acccccaac agaaactgg 
      gagtgtgaa agcaaaatt cccgaaaaa tcgaaacaa 
      tgcaaagac attgcgaga caattggtt gaggatgag 
      ccggggaga aatattaat gtagtgatg ggtggagga 
      agacagtgt ttacaatcg ggggttgaa gataaacca 
      ggtgatctt gttgataga tgggcatgt tatagttta 
      gatgggcga gatttgata agagactgg gagatggac 
      aaaaagaaa agaaataaa acttatgcc actgtgttg 
      aatagtgaa gaattgaag gcagtagac accagtaat 
      gaatttgtt ttaggaatt tttgcaaat ggacacatg 
      tcaatggat tacgcccga gataaatca ccaactgga 
      atgccttca ctcacgaac atgacacaa aaagcaatt 
      gaaattcta agccaaaac ccagaagga tatattttg 
      gtagtagaa ggtggtctt atcgatttt gcccatcac 
      agaggcaac gctagaaaa gctctagac gagactgtc 
      tcacttagt gatgcagtt cagatagct gtcgaaaaa 
      acggatcca cgagagaca ttaattatt gtaactagt 
      gatcactcg cattctttg gtatttact ggatatcct 
      tcaaggaac aaaagtgta ttagacatt gcacaaaag 
      tcaaaaatt gaccacata ccctacact tcgttacta 
      tatggaact ggaggtcca aacaattat caatactcc 
      gttattaac aataccatt tcaagacaa gatccttca 
      attaacaat acttcagat tttgaatat tcacaacaa 
      gctgtagtt ctaacggat gaagtgact catggtgga 
      agcgatgtt ttaatctat gctaaaggt ccgatgtct 
      catttattc cacagtgta catgaacag acgtatgtt 
      gcttacgta attagttac gccgcaaaa attggacca 
      tttaaaaat ggaagttca tcctttatc tacacgaaa 
      caaatttta aaatgcagc aagattgcg cttttcacg 
      cgactctat tccagccga aaccttcta aaattgaaa 
      gagcgcgga atgtaccaa gacattttc ccagataca 
      gcaacaaac acggttacg gatttatta aatgcagcc 
      ccacagcgt gtttatgca ggctttgac cccactgca 
      gacagcctt catatcgga aatttactt gttcttatt 
      aatttaata cactggcaa aggggtggc caccaaccc 
      atagcactg attggaggt gctaccgct aaaattggc 
      gatccgagt ggtaaaaca tcagaaaga gaaaaatta 
      gcaagtgct tttgttgat gagaatgtg attggaatt 
      agaaacaac attgaaacg cttttttat aatcatgaa 
      aaataccta tggaagaag aagggaaaa ctggctcct 
      ctaataatc gtgaataac gaatcttgg tatagtaaa 
      atgtcacca atcgatttg attggtggt gctggtcgt 
      tatttgcga atgggaaca ttattaagc agaacaagt 
      gttcaaaca cgccttcag tctcccgct ggtatgagt 
      tttaccgaa tttagttac caattattt caagcatat 
      gattggctg catttattt caaaaatac aattgctct 
      tttcaaatt ggtggaaat gatcaaatg ggcaatatt 
      atgtcgggg catgaactc atcagcaaa acgtgcgac 
      aaacaagtt tatgctctt actgttccg ataattaca 
      accgaaatg ggcgacaaa ttcggcaaa tctgcagga 
      aacgccatg tggctatct gctgaaaaa acctcacct 
      ttcactctt taccagttt ttcgtccgt caagttgac 
      accgaagtt gaaaaaatg ttaaagttc ttcacattt 
      gaaccagtc ggaagcatt aaagatctg atgagacgc 
      cacaccgaa aaacctgaa ttgcgacta cctcaaaaa 
      tatcttgct gagcaagtg gtgctctta gtgcatggg 
      gaaaagggt ttagaggct gcgcataaa gcgacaaaa 
      gttttatac gaacgaagc attacagca ctcgctgaa 
      atgaatcct ttggaaata aagagttta tttgaagga 
      gccacagtt gtggaaatt ttaccagag cctggacaa 
      agtgtgcta gatttggcg atgagcgct ggatgtttt 
      cttaccaaa gaagatgca ctgagaatc atttcagcc 
      ggcggtttt tacatcaac catcaaaaa acaaacaat 
      cctaatgaa gttttaaac ttaaatgta cataggtta 
      agcaataat gtaacgtta cttcgagta ggcaaaaag 
      aactattat gtagtaaaa tggctttca taataaaac 
      ttacgtttt gtaacttct tttttttta attacgcct 
      ggcggaaca gagaaaaac tggcttctc caaggtaag 
      cattttcaa aatatttta taatttttt cttcaaaat 
      agtttatta tcgttttta ataatgtta ttatagagt 
      t
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