Status
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Gene information
General information
TC033816
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.

TC030207,TC010044
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_007422.5: 9876386..9891465
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
    TC033816
    iBeetle screen iB_01645

    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: 5 (9 dpi)
    Days post injection


    adult lethality - Number of animals showing the phenotype: 3 (11 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: 60.0% (includes death as larva, prepupa, pupa)
    Lethalities 22 days after larval injection: 100.0% (includes death as larva, prepupa, pupa, adult)

    larva lethality - Number of animals showing the phenotype: 6 (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: 2 (16 dpi)
    Days post injection


    pupal molt delayed (still larva) - Number of animals showing the phenotype: 2 (16 dpi)
    Days post injection


    larva lethality late (L7) - Number of animals showing the phenotype: 2 (19 dpi)
    Days post injection


    pupal molt - (19 dpi)
    Days post injection


    iBeetle screen iB_07450

    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: 50.0% (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: 3 (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
    • >TC033816 RA
      MINLFAIIL PIDSTVRES KKFMHLCFK TQEYYFTED 
      AQKAEELAR LTRISKHYL REFSLAGFC NVSKSIIFC 
      FIGNVATYV IITLQFNEA NIEKSKYVD KDRTGKAIR 
      QKFLDYFIN KNNHNFIKS SPVVPFCDP TVAFVNAGM 
      NQFKGVFLG THQPQYKNV ANSQKCIRV GGKHNDLNV 
      VGGDGYHHT FFEMLGNWS FGDYFKQEA CELAWKLLT 
      EEYKIHPSR LYVTYFKGD PVLGIDEDL ETKEIWKTI 
      GVSEDRILP FTSKDNFWE MGPTGPCGP CTEIHIDHT 
      TSNNRAKLV NKGLHDLTE LWNLVFIQY NRKSNESVD 
      LLPEKHVDT GMGFERLTA VLQGKISNY DTDNFTYLL 
      KAIHKNCSK IPEYRGTFG EKDWNSLDA SYRVLADHA 
      RMVTVCLAD AMIPEQNQK LRRVMRRAF LLCENVFNK 
      DSGLLKELS NYVIEHLGC VYPEMERNV KQIHQIIDY 
      EEEVYKSVR KQATEEWVK LAKHQPKLL SLDINENPS 
      LVAGYKEIS STNPKFIDA QLAFKLYDT FGLDEDSIS 
      SLAQILNLP FNASDLHKQ LELAKIRSK EQGLVSGPN 
      VYEQLTRAK VPKTDRSTL YNYAKKGEK YVFDDLEVV 
      ILKIVKDSK LVRQIESEN FYGLVLDKS SFYTEAGGQ 
      ISDRGAINF GDCVFEVTE AQDCNGYIL HKGFLRSGE 
      SGRVKVGMT GSLKIDPDF RLKNMRNHT AVHLLNSVL 
      KKVKGATCQ KSSKVTQNY FNLDVGIFA EKLSLDELE 
      LVESTITKV IQGRLPVNI SELDSQRLA NLDEIILIP 
      GEVYPETGI RVVEVKGGE GFLSREPCC GTHVLNTSD 
      IEDFCIVSM KSLGRSTTS IHGVTGDRA KVAKSNGTH 
      LLTNLETLE KSVSDNIDK PDVLNMAIT SFKKQLNYS 
      VEDENILPV VVKQQCADI LNKISKQIK DSGKRNLRE 
      FIEIEMRNA MESNVSQTA KNRKYMVHY LRTSLILEN 
      VSLQKATNL CPDIPVLII SYSDNVVKA RCCVPENMK 
      NVSFDAEKW MKETVAAVF KCSLVPPKG YDPSLVCNM 
      KAKRVKLED WDPLLQESL ELAKKFAEQ NL
    CDS sequence
    • >TC033816 RA
      atgattaat ttgttcgcc attattcta cccatcgat 
      tcaactgta cgagaaagt aaaaagttc atgcacctg 
      tgcttcaaa actcaggaa tactacttt actgaagat 
      gctcaaaaa gctgaagaa ttagcaagg ttaacacgc 
      atctcaaag cattaccta cgcgagttc tctctagcc 
      ggcttttgt aatgtcagc aaatcaatt atcttttgc 
      tttattgga aatgtggca acttacgtt atcattaca 
      cttcagttt aatgaagca aatattgaa aagagtaaa 
      tatgtggac aaagacagg acgggtaaa gccatccga 
      caaaaattt ctcgattac ttcattaac aaaaacaac 
      cataacttc atcaaatca agcccggtg gtccctttt 
      tgcgacccc accgtggct tttgtgaat gccgggatg 
      aatcagttc aaaggagtt ttcttgggg acacaccaa 
      ccccaatac aaaaacgtt gcaaattca cagaagtgc 
      atccgagtc ggaggcaaa cacaacgac ttgaatgtc 
      gttggaggc gatggctat caccacacg ttttttgaa 
      atgttgggg aattggtcg tttggcgat tacttcaag 
      caagaggcg tgtgaattg gcttggaag ttgctaact 
      gaagagtat aaaatacac ccctcaagg ttgtatgtc 
      acatatttc aagggtgac ccagttttg gggatcgat 
      gaagatttg gaaaccaaa gaaatttgg aaaacaatt 
      ggggtgtca gaagacagg attttacct ttcacatca 
      aaagacaat ttctgggaa atggggcct acgggccct 
      tgtggccct tgtactgaa attcacatc gaccataca 
      acgtcaaat aatcgggcg aaactagtc aacaaggga 
      ctccatgat ctcactgaa ctctggaat ttagttttt 
      attcaatac aatcgtaaa agcaatgaa tcagttgat 
      ttactccct gaaaagcac gtcgatacc ggtatgggc 
      ttcgagaga ctcactgca gttcttcag ggcaagatt 
      agcaattat gacactgat aatttcact tatttactt 
      aaagcaata cataagaat tgcagcaaa atccccgaa 
      tatcggggc acgtttggt gaaaaggat tggaacagt 
      ttggatgca tcctacagg gtattggcc gatcacgca 
      agaatggtg accgtttgt ctcgctgat gcaatgatt 
      cctgaacag aaccaaaaa ctgagacgt gtaatgaga 
      agggcgttc ctattgtgt gaaaatgtt ttcaacaaa 
      gacagtggg ctgcttaag gagctctca aattacgta 
      atcgaacac ttggggtgt gtttacccc gaaatggaa 
      cgcaacgtc aaacagata caccaaatt atcgattat 
      gaagaggaa gtttacaaa tcggtgcgc aaacaagcc 
      acagaagag tgggtcaaa ctggccaaa caccaaccc 
      aagctcctt tcactcgac ataaacgaa aaccccagc 
      ctggtagcc gggtataaa gagatcagt tctaccaac 
      ccgaaattt atcgacgct caactcgct tttaaactc 
      tacgatacg ttcggtttg gacgaggac tcgatctcc 
      agtctggca caaatcttg aatttgccc ttcaacgcg 
      tccgacttg cacaagcaa ctcgagctc gccaaaatt 
      cggagcaag gaacagggc ctagtgtca ggtcctaac 
      gtctatgag caactcact agggctaaa gtaccgaaa 
      acagacagg tctactctt tacaattac gcaaagaag 
      ggcgagaag tacgtcttc gatgatctg gaagtggtc 
      attttgaaa atcgtcaaa gattcgaag ttagtccgc 
      caaatcgag tcggaaaat ttttacggg ttggttctg 
      gacaagtca agtttttac accgaggct ggagggcaa 
      atcagcgac aggggcgca attaatttt ggggactgc 
      gtattcgaa gtgactgaa gcacaagac tgtaacggg 
      tatatttta cataagggg tttctccgt tccggggag 
      agtgggcgt gttaaggtc ggcatgacc ggtagtttg 
      aaaattgac cccgatttt cgcctcaaa aatatgaga 
      aatcacacg gctgttcat ttgctgaac tcggtgtta 
      aaaaaggtg aaaggcgcc acgtgccag aagtcgagt 
      aaagtcacc cagaattat ttcaacttg gatgttggt 
      atttttgct gagaaattg tcgcttgat gagttagag 
      ttagtagag agtacgata acaaaagtt attcagggc 
      cgcctaccg gtcaatatc agcgagttg gatagtcag 
      aggttggcc aacttggat gagattatt ttaatccca 
      ggtgaggtg tacccggaa actgggatt agggttgtg 
      gaagttaag ggtggagag gggtttttg tcacgtgag 
      ccgtgctgt ggtacccat gttttgaat acaagtgat 
      attgaggac ttttgtatc gtcagtatg aagtcacta 
      gggaggagc accacttcc attcacggt gtcacggga 
      gatagggct aaagtggcc aaaagcaac ggaacccat 
      cttttaact aacttggaa accctcgag aaaagcgtt 
      tctgacaat attgataag cctgatgtg ttgaatatg 
      gcaattaca tcattcaag aaacaactg aattacagt 
      gttgaagac gagaacata ttgccggtt gttgttaaa 
      caacaatgt gcggatatt ttgaacaaa attagtaag 
      cagattaag gattcaggc aagagaaat ttacgagag 
      tttatcgaa atcgagatg cgcaacgct atggagtca 
      aatgtttca caaacagca aaaaatcgg aaatatatg 
      gtacactac ttgcgcaca tcacttatt ttggaaaac 
      gtttcgttg caaaaagcg acaaattta tgtcctgac 
      attcctgtt ttgataatt tcttactct gataatgta 
      gtaaaggcc cgttgttgt gttcctgag aatatgaaa 
      aatgtctcg tttgatgcc gaaaaatgg atgaaagag 
      accgtcgca gccgttttc aaatgttct cttgttcct 
      cccaagggt tatgacccg tctcttgtg tgcaatatg 
      aaagccaag cgagttaaa ttggaagat tgggaccct 
      ttattacag gaaagtctc gaattggct aagaaattc 
      gctgaacaa aatctgtag
    mRNA sequence
    • >TC033816 RA
      aacgaaaat tttttgcaa tgtcaagac gagaaattt 
      gaaagcttc gtttactac catcgaatt ttaggagaa 
      acggttgat ctttttaac agcttgttt ggctaccag 
      actttgtta atcacattt catattgga tcgcagctg 
      gtggtgtgc ctaaatttt tcttttggt tttctcact 
      ccttcaagc tttactcta cagttggat ctgttaata 
      agcaatttt tgggttgtg cttgtgatt atcttttca 
      caaacgcct cagtctcag aagtataaa taattgaag 
      cagttacgt ttttagttt aatggtata ctcgctagg 
      actcttaag tatttattt ttttcataa gccaaatta 
      gttattcta tccgcgtcc tcgccaatt agtaattag 
      acgtcataa cactcaatt tcctgaacg agagaacta 
      attaattaa aaatgtcat cataccgcg gtagaactc 
      taacgagcc aacatgaca agtttgaaa ctatcaacg 
      caagatatc aattttatt cgtcccttg tggaaattc 
      ttgaacgtt ttcctaatt acaccatgg tacgatttc 
      aacaaaaac tgtgttcac aaaccactt ttaatgaag 
      ttatgtgga tgtgcctta atcattgtt attatcttt 
      tgggctatt attccaaca ttcgacgac actgtaaga 
      cacatttat catataatg ccattctcg gaaaaatgc 
      gtcaacatt ttaacagct gcgaattta aaattcttc 
      gtttttttg gtcataata aagtgctct tttctggat 
      tccgacaag tgggttgtt ttgatgaca cattttcaa 
      tacatcgac gagaaatta aacaacaag ggaagaaaa 
      gaagagcag atattgaaa aacttttac ttgtggttc 
      ttcctgaaa caagtcctt tttctttta ttcgcattt 
      tacataacg tacatctgg tcactggtt ttgggttat 
      tcgtttatc aaatccctt ctaactgat attttacta 
      gagttgttt tatgcattg caagtcata attctgata 
      cgatgtctt gtcatttca attaaaatg cgataccaa 
      ggcctaaat aataagctc cgacaacta cactcaagt 
      ccgaacatt gtacaagaa ctgcggaaa cttgttgaa 
      tattttcga attttagga gaaatgatt gacttgttt 
      aacgattta ttcggatac ccaattatc ctatttatt 
      tttcacagt ggattacaa gtggttgtt tgtgtaaat 
      tttatgttt tcaatggtc aggcagact caggaatcc 
      acgttccat catttcata atcgcaaat gtctcggta 
      ttaattttt acgatgatt aatttgttc gccattatt 
      ctacccatc gattcaact gtacgagaa agtaaaaag 
      ttcatgcac ctgtgcttc aaaactcag gaatactac 
      tttactgaa gatgctcaa aaagctgaa gaattagca 
      aggttaaca cgcatctca aagcattac ctacgcgag 
      ttctctcta gccggcttt tgtaatgtc agcaaatca 
      attatcttt tgctttatt ggaaatgtg gcaacttac 
      gttatcatt acacttcag tttaatgaa gcaaatatt 
      gaaaagagt aaatatgtg gacaaagac aggacgggt 
      aaagccatc cgacaaaaa tttctcgat tacttcatt 
      aacaaaaac aaccataac ttcatcaaa tcaagcccg 
      gtggtccct ttttgcgac cccaccgtg gcttttgtg 
      aatgccggg atgaatcag ttcaaagga gttttcttg 
      gggacacac caaccccaa tacaaaaac gttgcaaat 
      tcacagaag tgcatccga gtcggaggc aaacacaac 
      gacttgaat gtcgttgga ggcgatggc tatcaccac 
      acgtttttt gaaatgttg gggaattgg tcgtttggc 
      gattacttc aagcaagag gcgtgtgaa ttggcttgg 
      aagttgcta actgaagag tataaaata cacccctca 
      aggttgtat gtcacatat ttcaagggt gacccagtt 
      ttggggatc gatgaagat ttggaaacc aaagaaatt 
      tggaaaaca attggggtg tcagaagac aggatttta 
      cctttcaca tcaaaagac aatttctgg gaaatgggg 
      cctacgggc ccttgtggc ccttgtact gaaattcac 
      atcgaccat acaacgtca aataatcgg gcgaaacta 
      gtcaacaag ggactccat gatctcact gaactctgg 
      aatttagtt tttattcaa tacaatcgt aaaagcaat 
      gaatcagtt gatttactc cctgaaaag cacgtcgat 
      accggtatg ggcttcgag agactcact gcagttctt 
      cagggcaag attagcaat tatgacact gataatttc 
      acttattta cttaaagca atacataag aattgcagc 
      aaaatcccc gaatatcgg ggcacgttt ggtgaaaag 
      gattggaac agtttggat gcatcctac agggtattg 
      gccgatcac gcaagaatg gtgaccgtt tgtctcgct 
      gatgcaatg attcctgaa cagaaccaa aaactgaga 
      cgtgtaatg agaagggcg ttcctattg tgtgaaaat 
      gttttcaac aaagacagt gggctgctt aaggagctc 
      tcaaattac gtaatcgaa cacttgggg tgtgtttac 
      cccgaaatg gaacgcaac gtcaaacag atacaccaa 
      attatcgat tatgaagag gaagtttac aaatcggtg 
      cgcaaacaa gccacagaa gagtgggtc aaactggcc 
      aaacaccaa cccaagctc ctttcactc gacataaac 
      gaaaacccc agcctggta gccgggtat aaagagatc 
      agttctacc aacccgaaa tttatcgac gctcaactc 
      gcttttaaa ctctacgat acgttcggt ttggacgag 
      gactcgatc tccagtctg gcacaaatc ttgaatttg 
      cccttcaac gcgtccgac ttgcacaag caactcgag 
      ctcgccaaa attcggagc aaggaacag ggcctagtg 
      tcaggtcct aacgtctat gagcaactc actagggct 
      aaagtaccg aaaacagac aggtctact ctttacaat 
      tacgcaaag aagggcgag aagtacgtc ttcgatgat 
      ctggaagtg gtcattttg aaaatcgtc aaagattcg 
      aagttagtc cgccaaatc gagtcggaa aatttttac 
      gggttggtt ctggacaag tcaagtttt tacaccgag 
      gctggaggg caaatcagc gacaggggc gcaattaat 
      tttggggac tgcgtattc gaagtgact gaagcacaa 
      gactgtaac gggtatatt ttacataag gggtttctc 
      cgttccggg gagagtggg cgtgttaag gtcggcatg 
      accggtagt ttgaaaatt gaccccgat tttcgcctc 
      aaaaatatg agaaatcac acggctgtt catttgctg 
      aactcggtg ttaaaaaag gtgaaaggc gccacgtgc 
      cagaagtcg agtaaagtc acccagaat tatttcaac 
      ttggatgtt ggtattttt gctgagaaa ttgtcgctt 
      gatgagtta gagttagta gagagtacg ataacaaaa 
      gttattcag ggccgccta ccggtcaat atcagcgag 
      ttggatagt cagaggttg gccaacttg gatgagatt 
      attttaatc ccaggtgag gtgtacccg gaaactggg 
      attagggtt gtggaagtt aagggtgga gaggggttt 
      ttgtcacgt gagccgtgc tgtggtacc catgttttg 
      aatacaagt gatattgag gacttttgt atcgtcagt 
      atgaagtca ctagggagg agcaccact tccattcac 
      ggtgtcacg ggagatagg gctaaagtg gccaaaagc 
      aacggaacc catctttta actaacttg gaaaccctc 
      gagaaaagc gtttctgac aatattgat aagcctgat 
      gtgttgaat atggcaatt acatcattc aagaaacaa 
      ctgaattac agtgttgaa gacgagaac atattgccg 
      gttgttgtt aaacaacaa tgtgcggat attttgaac 
      aaaattagt aagcagatt aaggattca ggcaagaga 
      aatttacga gagtttatc gaaatcgag atgcgcaac 
      gctatggag tcaaatgtt tcacaaaca gcaaaaaat 
      cggaaatat atggtacac tacttgcgc acatcactt 
      attttggaa aacgtttcg ttgcaaaaa gcgacaaat 
      ttatgtcct gacattcct gttttgata atttcttac 
      tctgataat gtagtaaag gcccgttgt tgtgttcct 
      gagaatatg aaaaatgtc tcgtttgat gccgaaaaa 
      tggatgaaa gagaccgtc gcagccgtt ttcaaatgt 
      tctcttgtt cctcccaag ggttatgac ccgtctctt 
      gtgtgcaat atgaaagcc aagcgagtt aaattggaa 
      gattgggac cctttatta caggaaagt ctcgaattg 
      gctaagaaa ttcgctgaa caaaatctg tagtagaga 
      aataagatt gtttttttt taataaaca cttatttat 
      ttt
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