Status
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Gene information
General information
TC034448
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.

TC012763,TC012762
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: 14323847..14372077
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
    TC034448
    Gene ontology

    GO terms for Tribolium

    • {{ goTerm.term }}
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      Status: {{ goTerm.status }}
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      Quotation: {{ goTerm.quotation }}
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    There are no GO terms for Tribolium at the moment. You can help the community by providing some GO terms for this gene.


    GO terms for the homologous Drosophila genes


    Assign a GO term to this Tribolium gene by filling in the fields. Repeat to add several GO terms. Search AmiGO for the correct GO ID. Use only the most specific term - use "graph views" to browse related terms ("child terms" are more specific; the more general "parent" terms will be automatically linked). Only information based on Tribolium data should be entered - do not define terms just based on Drosophila knowledge. We will review this information and submit the annotation to the Gene Ontology consortium.


    Gene
    {{ goGene }}
    The Tribolium gene the GO term is assigned to.
    GO ID *
    {{ blurIdResult }}
    The ID of the GO term you assign to the gene. Please enter only the ID of the GO term with proceeding 'GO:'. This information is mandatory.
    Evidence
    {{ blurEvidenceResult }}
    The evidence code describes with which method the function or location, expressed by the GO term, was show for this gene. Please refer to this page for a definition of the evidence code and a decision tree.
    Reference *
    {{ blurReferenceResult }}
    The reference where the function or location is described for the gene. Please enter a PubMed ID. This information is mandatory.
    Quotation
    If you paste here the relevant section of the reference it is for us a great help to review your annotation. The quotation may be also be displayed to the users of iBeetle-Base.
    Name
    You may enter your name. The contact information can help us to clarify any questions. If you agree, we may show your name on this page to honor your contribution.
    Email
    We will use your email address only for possible questions regarding this annotation. We will never show your address or give it away.
    Publish name
    If you have entered your name and agree by marking the checkbox. we might show your name on this page to honor your contribution.
      {{ unavailableText }}

    * : mandatory fields
    For help on how to submit larger datasets or non experimental data please contact us.
    iBeetle screen iB_02017

    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: 2 (3 dpi)
    Days post injection


    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: 22.22% (includes death as larva, prepupa, pupa)
    Lethalities 22 days after larval injection: 50.0% (includes death as larva, prepupa, pupa, adult)

    larva lethality - Number of animals showing the phenotype: 2 (11 dpi)
    Days post injection


    eclosion not fulfilled (died as pupa) - Number of animals showing the phenotype: 1 (11 dpi)
    Days post injection


    Pupal morphology
    elytra proximal partiallyblistered - Number of animals showing the phenotype: 7

    Adult morphology
    gula surface irregular - Number of animals showing the phenotype: 4


    elytra partially blistered - Number of animals showing the phenotype: 4


    hindwing shape irregular - Number of animals showing the phenotype: 4

    Fertility
    egg production: number <50% of expected -
    vitellogenic egg chamber slightly number decreased - Number of animals showing the phenotype: 2

    tropharium slightly shape irregular - Number of animals showing the phenotype: 2

    ovary without detectable defects - Number of animals showing the phenotype: 1
    iBeetle screen iB_05313

    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)

    pupa eclosion not fulfilled (died as pupa) - Number of animals showing the phenotype: 1 (3 dpi)
    Days post injection


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


    Fertility
    egg chamber not present - Number of animals showing the phenotype: 3

    egg chamber mostly not present - Number of animals showing the phenotype: 1

    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: 50.0% (includes death as larva, prepupa, pupa)
    Lethalities 22 days after larval injection: 90.0% (includes death as larva, prepupa, pupa, adult)

    larva lethality early (L5/6) - Number of animals showing the phenotype: 5 (11 dpi)
    Days post injection


    pupal molt delayed (still larva) - Number of animals showing the phenotype: 4 (11 dpi)
    Days post injection


    larva lethality late (L7) - Number of animals showing the phenotype: 4 (16 dpi)
    Days post injection


    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC034448 RA
      MGLGVSKEA PCPELERPE SVISSKLHG RQASIRQST 
      MRAKSKQPM PDPNELERR FTKVLASMD LPPDKAKLL 
      KNYDSEKKW DIICDQEMV HAKDPPSFY LNKLRTYLD 
      PKASRSHRK RKMVGESTS TQVLRDLEI SLRTNHIEW 
      VREFLNEEN NGLDVLIDY LSFRLGMMR HEQRIAESR 
      TASEERLGP GQPPPAPSV QNGYIRAPL DIPDSSSVK 
      RRSKHVAKL NMGESKDDI HVCIMCMRA IMNNKYGFN 
      MVIQHREAI NCISLSLIH KSLRTKALV LELLAAICL 
      VKGGHEIIL SAFDNFKEV CHEEHRFQT LMDYFLNYD 
      VFHIEFMVA CMQFVNIIV HSVEDMNFR VHLQYEFTA 
      LGLDSYLEK LRFTESEEL QVQISAYLD NVFDVAALM 
      EDSETKTAA LEKVAEMED ELAQLHEKI QDIEHQALE 
      EIAHLKTDL AQLRQERDE LAEFKRHAD EEVNTLRRV 
      VQHHEQESK NRQSMFESK IAELEKTVP KSASVAGIS 
      DLTPPDSTT SPVPASPVI PPPAAPPPP PCPPPPPSR 
      VPSAPVPPP PAGFMQAPD GAMTIKRKV QTKYKLPTI 
      NWIALKPNQ VRGTIFNEL DDDKLHGFI DFNDFEERF 
      KIGNAGLLA NGTSELDGL STFPSKRFK KPENVTLLE 
      HTRLRNIAI SRRKLEMSP EKVINAINL LDLKQLSLE 
      NVELLQRMV PTEQETKAY REYVNDKKN VNLLTEEDK 
      FLLQLTKVE RISAKLSIM SYIGNFYDN LHLITPQTH 
      AIISGSNSV KNSKKLRSV LEIILAFGN YLNSSKRGP 
      AYGFKLQSL DTLLDTKST DKRMCLLHY IVATIRQKF 
      PELLNFDSD LHYIEKAAV VSLENITTD VHELEKGME 
      AVRKEAEIR GKGPQSIVV RDFLANAEE KLRRLRNDA 
      KTAQEAFRE CVEYFAESP RTTDANTFF SLLVRFVKA 
      FKQADQDNE QRRRLEIAA QNANNKTNE DVAKRNKIN 
      QKKQQEAVI NELKHKTRL VQEKKLLQQ DEVYNGALE 
      DILLGLKSE PYRRADAVR RSQRRRIDN NRLSKTMEE 
      LEV
    CDS sequence
    • >TC034448 RA
      atgggcttg ggcgtatcg aaggaggcg ccttgcccg 
      gagctggaa aggccggaa agtgtgatc tcgtcgaaa 
      ttgcacgga agacaggcg agcatccgc cagagcacc 
      atgagggcc aagtcgaag cagcccatg cccgacccc 
      aacgaactg gagaggaga ttcaccaaa gtgctggcc 
      tccatggac ttgccgccc gataaagcc aaactgctg 
      aaaaactac gattcggag aagaaatgg gacatcatc 
      tgcgatcag gaaatggtg cacgctaaa gaccctccc 
      tccttctac ttaaacaaa ctccggacg tacctcgac 
      cccaaggcc tcccgcagc cacaggaag cgaaaaatg 
      gtcggcgag tcgacttcg actcaagtc ctgcgtgac 
      ctggagata tccctgcgg acgaaccac atcgaatgg 
      gtgcgagaa tttttaaac gaggagaac aacggcttg 
      gacgtcctg atcgactat ttgagcttt aggctgggc 
      atgatgcgg cacgagcag cgcatcgcc gagtccagg 
      accgcctcc gaggagcgg ctgggcccc ggccagccg 
      ccgcccgcc cccagcgtc cagaacgga tacattcgc 
      gcccccttg gacatcccg gacagctcg agcgtcaaa 
      cggcgctcc aagcacgtc gccaagctg aacatgggg 
      gagtccaag gacgacata cacgtctgc atcatgtgc 
      atgagggcc attatgaac aataagtat gggttcaat 
      atggtgatc cagcaccgt gaggccatc aactgcatt 
      tcgctcagc ttgatccac aagagtttg cgcacgaag 
      gcgctggtg ctggagctt ttggcggcg atttgcctg 
      gtcaaaggc gggcacgag ataattctc tccgcgttt 
      gataatttc aaagaggtg tgtcatgag gagcaccgc 
      ttccagacg ttgatggat tattttctc aattacgat 
      gtcttccac atcgagttt atggttgcg tgcatgcag 
      tttgtgaac attattgtg cattcggtg gaggatatg 
      aactttagg gtccatctg cagtatgag ttcacggcg 
      ctggggctg gacagctat ttggagaag ttgaggttt 
      acggaaagt gaggagttg caagtgcag atctcggcc 
      tatttggac aacgtgttc gatgtggcg gcgctcatg 
      gaggatagc gagaccaag acggcggcc ttggagaag 
      gtggccgag atggaggac gagctggcg caactccac 
      gaaaaaatc caagacatc gaacaccaa gccctcgaa 
      gagatcgcc cacttgaaa accgacctg gcccaactg 
      cgccaagaa cgcgacgaa ctcgccgaa ttcaaacgc 
      cacgccgac gaagaggtc aacacttta cgtcgcgtt 
      gtccaacac cacgaacaa gaatcgaaa aatcgccaa 
      tcgatgttc gaatcaaaa atcgccgaa ctcgaaaaa 
      acagtcccg aagtcggcc tcagttgcc ggtatttcc 
      gacctgacg ccgcccgat tcgaccacc tcgcctgtg 
      cccgcctcg cctgtgata ccgcccccg gccgcgccg 
      ccgccgccg ccgtgcccg ccgccgccg ccgtccaga 
      gtgccttcc gcgcccgtg ccgccgccg cccgccggc 
      ttcatgcaa gcgccagat ggcgccatg accatcaaa 
      cgcaaagta caaaccaaa tacaaactg ccaaccatc 
      aattggatc gcactgaag ccgaatcaa gtcagaggg 
      acgattttt aacgaattg gacgatgat aagctgcac 
      ggattcatc gattttaac gactttgag gagcgattc 
      aagattgga aatgccggc ttgttggcg aatggcact 
      agtgagctt gatgggctt tcgacgttc ccgagcaaa 
      aggttcaaa aagccggaa aatgtcaca cttttggag 
      cacaccaga ctgagaaat atcgcaatt tcgaggcga 
      aaattggaa atgtcgcct gagaaagtg ataaatgcg 
      atcaattta ctcgatttg aaacaattg agtctcgaa 
      aatgtcgaa ttgttgcaa aggatggtg ccgaccgag 
      caggaaaca aaagcctat cgggagtat gtcaatgat 
      aagaaaaac gtcaatttg ttgactgaa gaggacaag 
      tttttgcta cagttgacc aaggtcgag cgaatttcg 
      gccaagttg tcgattatg agttacata ggcaatttt 
      tacgacaat ctgcacttg ataacgccg caaacacat 
      gcgatcatt tctggctca aattctgtg aaaaactcg 
      aagaaattg agatcagtt ttggagata atcctcgca 
      ttcggcaat tatctcaac agcagtaaa cgagggccg 
      gcttacggc ttcaaactt caatcactc gacacactt 
      ttagacacg aaatcgacc gataaaaga atgtgtctt 
      ttgcattac atcgtcgca acgattcgc caaaaattc 
      cccgaattg ctaaacttc gactcggac ctgcactac 
      atagagaaa gccgcagtt gtttctctc gaaaacatc 
      acaactgat gtccatgag ctagagaaa gggatggaa 
      gcggtccga aaagaggcc gaaatccgg ggcaaaggg 
      cctcagagc atcgtcgtt cgtgatttc ctcgccaat 
      gccgaggag aagttgagg cgattgagg aacgacgcc 
      aaaacagca caagaggcc ttcagggag tgcgtggag 
      tattttgca gagtcgcca aggaccacc gatgccaac 
      actttcttc tccttgtta gtcagattt gtcaaagct 
      ttcaagcaa gcggatcag gacaacgag caaagaagg 
      agactggaa atcgctgct caaaatgcc aacaataaa 
      accaacgaa gacgtggcg aagaggaat aaaatcaac 
      cagaagaaa caacaagaa gctgtaata aacgaattg 
      aaacacaaa acaagactg gtgcaggag aaaaagctc 
      ttgcaacaa gacgaagtg tacaatggt gcccttgaa 
      gatatcctc ctagggcta aaaagtgag ccttaccgt 
      cgtgccgat gcggtgcga agaagtcaa agacggagg 
      attgataat aatcgcttg tctaaaaca atggaagag 
      ttggaagtt tga
    mRNA sequence
    • >TC034448 RA
      ctttttttg cgaatgcgc tattacata acctcacag 
      tgtttacaa acaaaaaat tgtaaaaac ttatcaaaa 
      cacggaaga taagtcaac aaatcgcaa aaaatggat 
      caacaaaca acccccaac ggttccggg ttcgtttat 
      ttcaacacg agttcaaat caagaatat caaaaccaa 
      tcggacacg gctgacttt ttggccttt actgacact 
      ccccaagcg tctcctccc actaacaag tttaaacct 
      gcttactac agcagcccc cagtcgaaa ttccacagg 
      gggagcccc cgacaatac ccccaaaat aatcggagg 
      tatcagcgt tggtcgggg ggctataac cggtcttac 
      aatagctca aactccagt tttaaccag agtggctca 
      tttagggag agtgaccaa gagggccct aatatctta 
      aaattcata catccgtcg tttctggag gacccctgg 
      gcctccttg gaggccaaa ttgaaggaa aaggggcag 
      tttgacgat tcgtcgcaa tcagggtcc caggaaact 
      tagcaattt tagcgtcaa aaaaaaata caaattgtg 
      caaattaat taattttaa ttgtaggtt taaaaaatt 
      gatatttta ttaaaaaca cagttaaga aaatagttt 
      ttgagcgac agattcctg tattaccgg cccagtatc 
      ccgtttttt gtgtcaaga tggcgcccg aatcccaat 
      gtaaaaatt ccatcccaa acatgcgca catctctct 
      tggaaaatc taaaacttt gtttagttt tgcgagatt 
      tacgtttgc ggcgcgttg tgtgaaaac ctgttgcgt 
      ttgtgtgac gcgatcgtg cgtgatgcg gcaaagaac 
      gtgattttt ggtggcgca tagtgtatt ttttctgcg 
      accatgggc ttgggcgta tcgaaggag gcgccttgc 
      ccggagctg gaaaggccg gaaagtgtg atctcgtcg 
      aaattgcac ggaagacag gcgagcatc cgccagagc 
      accatgagg gccaagtcg aagcagccc atgcccgac 
      cccaacgaa ctggagagg agattcacc aaagtgctg 
      gcctccatg gacttgccg cccgataaa gccaaactg 
      ctgaaaaac tacgattcg gagaagaaa tgggacatc 
      atctgcgat caggaaatg gtgcacgct aaagaccct 
      ccctccttc tacttaaac aaactccgg acgtacctc 
      gaccccaag gcctcccgc agccacagg aagcgaaaa 
      atggtcggc gagtcgact tcgactcaa gtcctgcgt 
      gacctggag atatccctg cggacgaac cacatcgaa 
      tgggtgcga gaattttta aacgaggag aacaacggc 
      ttggacgtc ctgatcgac tatttgagc tttaggctg 
      ggcatgatg cggcacgag cagcgcatc gccgagtcc 
      aggaccgcc tccgaggag cggctgggc cccggccag 
      ccgccgccc gcccccagc gtccagaac ggatacatt 
      cgcgccccc ttggacatc ccggacagc tcgagcgtc 
      aaacggcgc tccaagcac gtcgccaag ctgaacatg 
      ggggagtcc aaggacgac atacacgtc tgcatcatg 
      tgcatgagg gccattatg aacaataag tatgggttc 
      aatatggtg atccagcac cgtgaggcc atcaactgc 
      atttcgctc agcttgatc cacaagagt ttgcgcacg 
      aaggcgctg gtgctggag cttttggcg gcgatttgc 
      ctggtcaaa ggcgggcac gagataatt ctctccgcg 
      tttgataat ttcaaagag gtgtgtcat gaggagcac 
      cgcttccag acgttgatg gattatttt ctcaattac 
      gatgtcttc cacatcgag tttatggtt gcgtgcatg 
      cagtttgtg aacattatt gtgcattcg gtggaggat 
      atgaacttt agggtccat ctgcagtat gagttcacg 
      gcgctgggg ctggacagc tatttggag aagttgagg 
      tttacggaa agtgaggag ttgcaagtg cagatctcg 
      gcctatttg gacaacgtg ttcgatgtg gcggcgctc 
      atggaggat agcgagacc aagacggcg gccttggag 
      aaggtggcc gagatggag gacgagctg gcgcaactc 
      cacgaaaaa atccaagac atcgaacac caagccctc 
      gaagagatc gcccacttg aaaaccgac ctggcccaa 
      ctgcgccaa gaacgcgac gaactcgcc gaattcaaa 
      cgccacgcc gacgaagag gtcaacact ttacgtcgc 
      gttgtccaa caccacgaa caagaatcg aaaaatcgc 
      caatcgatg ttcgaatca aaaatcgcc gaactcgaa 
      aaaacagtc ccgaagtcg gcctcagtt gccggtatt 
      tccgacctg acgccgccc gattcgacc acctcgcct 
      gtgcccgcc tcgcctgtg ataccgccc ccggccgcg 
      ccgccgccg ccgccgtgc ccgccgccg ccgccgtcc 
      agagtgcct tccgcgccc gtgccgccg ccgcccgcc 
      ggcttcatg caagcgcca gatggcgcc atgaccatc 
      aaacgcaaa gtacaaacc aaatacaaa ctgccaacc 
      atcaattgg atcgcactg aagccgaat caagtcaga 
      gggacgatt tttaacgaa ttggacgat gataagctg 
      cacggattc atcgatttt aacgacttt gaggagcga 
      ttcaagatt ggaaatgcc ggcttgttg gcgaatggc 
      actagtgag cttgatggg ctttcgacg ttcccgagc 
      aaaaggttc aaaaagccg gaaaatgtc acacttttg 
      gagcacacc agactgaga aatatcgca atttcgagg 
      cgaaaattg gaaatgtcg cctgagaaa gtgataaat 
      gcgatcaat ttactcgat ttgaaacaa ttgagtctc 
      gaaaatgtc gaattgttg caaaggatg gtgccgacc 
      gagcaggaa acaaaagcc tatcgggag tatgtcaat 
      gataagaaa aacgtcaat ttgttgact gaagaggac 
      aagtttttg ctacagttg accaaggtc gagcgaatt 
      tcggccaag ttgtcgatt atgagttac ataggcaat 
      ttttacgac aatctgcac ttgataacg ccgcaaaca 
      catgcgatc atttctggc tcaaattct gtgaaaaac 
      tcgaagaaa ttgagatca gttttggag ataatcctc 
      gcattcggc aattatctc aacagcagt aaacgaggg 
      ccggcttac ggcttcaaa cttcaatca ctcgacaca 
      cttttagac acgaaatcg accgataaa agaatgtgt 
      cttttgcat tacatcgtc gcaacgatt cgccaaaaa 
      ttccccgaa ttgctaaac ttcgactcg gacctgcac 
      tacatagag aaagccgca gttgtttct ctcgaaaac 
      atcacaact gatgtccat gagctagag aaagggatg 
      gaagcggtc cgaaaagag gccgaaatc cggggcaaa 
      gggcctcag agcatcgtc gttcgtgat ttcctcgcc 
      aatgccgag gagaagttg aggcgattg aggaacgac 
      gccaaaaca gcacaagag gccttcagg gagtgcgtg 
      gagtatttt gcagagtcg ccaaggacc accgatgcc 
      aacactttc ttctccttg ttagtcaga tttgtcaaa 
      gctttcaag caagcggat caggacaac gagcaaaga 
      aggagactg gaaatcgct gctcaaaat gccaacaat 
      aaaaccaac gaagacgtg gcgaagagg aataaaatc 
      aaccagaag aaacaacaa gaagctgta ataaacgaa 
      ttgaaacac aaaacaaga ctggtgcag gagaaaaag 
      ctcttgcaa caagacgaa gtgtacaat ggtgccctt 
      gaagatatc ctcctaggg ctaaaaagt gagccttac 
      cgtcgtgcc gatgcggtg cgaagaagt caaagacgg 
      aggattgat aataatcgc ttgtctaaa acaatggaa 
      gagttggaa gtttgagtc tcaaaagct gtaaataat 
      tcgtgtaaa tgctctctg atgctgatg atgatatta 
      agatgtgat tgtatttaa atatttttg tacatagtt 
      gcaatttta taaagtact actagtggt aaggctatt 
      tttatattg ttgtttgta cactgaaat aatcattac 
      ggcacattc cgggtttgt cgtttgtgc gtgttttac 
      tgatttgtg attttgaaa tgttttaat gcgaattgt 
      atgtttatt atttatagc caagcaata aaaagttta 
      ttattgatt gattaaaag ttgggaatt atttcgtgc 
      cttttccct cctttgaaa cttttccct tttttctat 
      tttagcttt tgttgtcaa acaaaaaaa cgaacatac 
      ggacttttt ttgactttt aacaactgt agaggaacg 
      taaggacgt aacaagtcc aaaaaccta cgatagaac 
      gagttttaa aaaaaattt tttttcacc tttttgaag 
      gtaaaagtg tattgaaat ttaagcaaa aaaattgaa 
      aattttaaa tctcgtgaa aagttggta taatacaga 
      aaatgaaca gctgaattc aatggaaca aaccgcatt 
      gcgctacga cttttagtt tttgagtta tgaattttt 
      ttaatgaat aaaagtttc cactataga tattgaaca 
      gctcgagaa ggttgtaaa ggcatataa aagtccata 
      aaactatgt tagaatgag ttaaaaaaa atctttttt 
      catttttgt gaaggccat taaaatatt aaacaaaaa 
      aatttagat cgggccaat tgacacttt tgtactttg 
      aaaggttca aaatggcga aaaacgttt ttggggcga 
      aaaagaaaa aataccttc ttgaccaac ttttaaaaa 
      ttggggttg tcggggatg cattgaaag gcatttaaa 
      ttatttaaa catttttat tgtaaaatg acaaagaat 
      aaatctaaa cttttcaga atcataata tgtttgata 
      taggtagta ctcgaatac atgatacag attgtaaaa 
      actgaacgc aagtttcaa aataatatt aaaaatata 
      taaacgcat cacttcgaa ctggccgtg tcattgtac 
      atataatga atactgctg gaacgagcc tcagccgct 
      gaaggaggt ctcgatcgt gtctcggcc gccttgaaa 
      caacggtgc aaaatagaa tttcgtact gtacgtaat 
      ttgataaaa atccaaatt gtgcccata gtttaagct 
      aaactttac caactacgt taatctaaa agtaatcta 
      agtatgtaa atagtagag tatatgtgg atagataga 
      tatacgcat cagctatgt cgatattta tgttgaatg 
      ca
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