Status
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Gene information
General information
TC034060
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.

TC030079,TC030080,TC005744
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: 8252529..8259600
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
    TC034060
    iBeetle screen iB_02900

    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)

    adult lethality - Number of animals showing the phenotype: 3 (9 dpi)
    Days post injection


    Analysis of embryonic musculature and early development
    number of eggs/embryos/larvae on slide: > 50 larval musculature pattern irregular - Penetrance (percentage of animals/eggs on slide): less than 30%

    affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): less than 30%
    remarks: muscle pattern potentially obscured by segmentation defect,
    Analysis of larval stage 1 cuticle
    number of eggs/embryos/larvae on slide: > 50
    embryo/egg without visible larval cuticle in cuticle preparation - Penetrance (percentage of animals/eggs on slide): less than 30%
    larva dorsal bent - Penetrance (percentage of animals/eggs on slide): less than 30%

    larva appendages shape irregular - Penetrance (percentage of animals/eggs on slide): less than 30%

    affected eggs/embryos/larvae total percentage of affected eggs/embryos/larvae on slide - Penetrance (percentage of animals/eggs on slide): 30% - 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: 0.0% (includes death as larva, prepupa, pupa)
    Lethalities 22 days after larval injection: 0.0% (includes death as larva, prepupa, pupa, adult)

    iBeetle screen iB_04006

    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: 10.0% (includes death as pupa, adult)

    eclosion not fulfilled (with pupal and adult features) - Number of animals showing the phenotype: 1 (3 dpi)
    Days post injection


    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: 0.0% (includes death as larva, prepupa, pupa)
    Lethalities 22 days after larval injection: 0.0% (includes death as larva, prepupa, pupa, adult)

    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC034060 RA
      MSIFLFFIL AVAIATPKK PDLKNIQDY GFSATLYKE 
      LGKDCHNFV ISPLLVETA FAALYLGSS GKTAEEIRT 
      VFHLPPKTG IEHKYKTLI PQLTNPGYT LNIANRVSL 
      AQKYKFVPT WETQIKNIF EVCADLLDM TNKNAAAEK 
      VNKWFRDKT NNKIDKMVS ADALTSETA LLLLSSLYF 
      QGNWDQQFS VSSTRKQTF YNHGKQQVS VDMMFASAR 
      SVNYGKSDL LNAQFLEIA FEKHHASMV VVLPDQLDG 
      IYQLETKIN EIFNFRDFC PETVNIWLP KFRMDSTFN 
      IDKILQQLG VKDAFDEEK AQFSGTVSC ADCHSPYIG 
      KVTQKTYLQ INERGVEAA AGQVVHVRV PLSGGRKQF 
      KADHPFLFY IKIKEVVVF IGRFREKTN NKIDKMVSA 
      DALTSETAL LLLSSLYFQ GNWDQQFSV SSTRKQTFY 
      NHGKQQVSV DMMFASARS VNYGKSDLL NAQFLEIAF 
      EKHHALMVV VLPDQLDGI YQLETKISE IFNFRDFCP 
      GTVNIWLPK FRMDSTFNI DKILQQLGV KDAFDETKA 
      QFSGTVSCA DCHSPYIGK VTQKTYLQI NERGVEAAA 
      GQVVHVRVP LSGGRKQFK ADHPFLFYI KIKEVVVFI 
      GRVLIKPDV FIICSIPGK KMHFLLFLT LCFASVSPY 
      TEIHDKKFA ASLYQELAK SCNNFVISP FLIETAFAA 
      LSLGSSRKT AGEFRTVFH LPQTHEAIK GRYKHFFSL 
      FTNKGYKFQ TASRISLLD ELELAQKWQ KQVRDIFNI 
      QAEVLDVTN KNASAEKID SWFREKTYN KVDKIVTPK 
      DLNVETGFF LLSASHFGA CWAKEFASK RTHKDNFYN 
      SGKLTQKTS VDTMYLLGY FNYGQSKQL EAKFLELNF 
      IEQQASMVI ILPDKIDGI QKLETKMDQ VLQFRDMRT 
      TAVDISLPK FRMDSHFDL IKFLQKLGI KQAFIDGKA 
      SFDGTVETD CDLPYITKV TQKTSMEIN ERGVEATAD 
      PAHAAPPSV APSYVKFKA DHPFIFYIK IKEVVVYVG 
      RVLAF
    CDS sequence
    • >TC034060 RA
      atgagtatt ttcctgttt tttatacta gctgtcgca 
      attgcgaca ccaaaaaaa ccagacctt aaaaatata 
      caagactat ggcttttcc gcaactttg tacaaggag 
      ttgggaaaa gattgtcac aactttgtc ataagtccc 
      ttgttggtt gagacagct ttcgctgcc ctctatttg 
      ggaagctct ggcaaaaca gctgaagaa atacgcacc 
      gtgtttcac ttacctcca aaaactggc attgaacac 
      aagtataaa acactcatc ccgcagtta accaatccc 
      ggctataca ctaaacata gcaaacagg gtttctcta 
      gcccaaaaa tacaaattt gtaccgact tgggagacg 
      caaatcaag aacattttc gaagtgtgt gctgatctt 
      ttggatatg acgaataaa aacgcagct gcggaaaaa 
      gtcaataaa tggtttcga gacaaaacg aacaataaa 
      atcgacaaa atggtgtcc gctgatgct ttgacaagt 
      gagacagct cttttgctt ttgagctcg ttgtatttc 
      caaggcaac tgggatcaa cagttttct gtatctagc 
      acacgtaaa cagacgttc tacaaccat ggaaaacag 
      caagtctca gtcgatatg atgtttgca tcagctcgg 
      tccgttaat tacggaaag agcgatctt ttaaacgct 
      caattcttg gaaatagcg tttgaaaaa caccacgct 
      tcgatggtt gtagttctt ccagatcaa ctggacggc 
      atttaccaa cttgaaacg aagattaat gaaattttc 
      aattttcgc gacttttgt cccgagacg gttaatatt 
      tggctgccg aaatttcgc atggatagc actttcaac 
      atcgacaag attctacaa caattgggt gtgaaagat 
      gcctttgac gaggaaaaa gcccaattc agtggaact 
      gtgtcatgt gccgattgt cactcgccg tacatcggc 
      aaagttact caaaaaact tatctgcaa atcaatgaa 
      cgaggagtt gaagcagca gctggtcaa gtagtacat 
      gttcgtgtt ccgttgtct ggtgggcgc aagcaattt 
      aaagccgac catcctttc ctgttttat atcaagata 
      aaagaagtt gtggtcttt attggacga tttcgggag 
      aaaacgaac aataaaatc gacaaaatg gtgtccgct 
      gatgctttg acaagtgag acagctctt ttgcttttg 
      agctcgttg tatttccaa ggcaactgg gatcaacag 
      ttttctgta tctagcaca cgtaaacag acgttctac 
      aaccatgga aaacagcaa gtctcagtc gatatgatg 
      tttgcatca gctcggtcc gttaattac ggaaagagc 
      gatctttta aacgctcaa ttcttggaa atagcgttt 
      gaaaaacac cacgctttg atggttgta gttcttcca 
      gatcaactg gacggcatt taccaactt gaaacgaag 
      attagtgaa attttcaat tttcgcgac ttttgtccc 
      gggacggtt aatatttgg ctgccgaaa tttcgcatg 
      gatagcact ttcaacatc gacaagatt ctacaacaa 
      ttgggcgtg aaagatgcc tttgacgag acaaaagct 
      caattcagt ggaactgtg tcatgtgcc gattgtcac 
      tcgccgtac atcggcaaa gttacccaa aaaacttat 
      ctgcaaatc aatgaacga ggagttgaa gcagcagct 
      ggtcaagta gtacatgtt cgtgttccg ttgtctggt 
      gggcgcaag caatttaaa gccgaccat cctttcctg 
      ttttacatc aagataaaa gaagttgtg gtctttatt 
      ggacgagtt ttaatcaaa ccagacgtg tttattatt 
      tgctcaatc ccgggcaaa aaaatgcat tttctctta 
      tttttgact ttgtgcttc gcatctgta tccccatac 
      accgaaata catgacaaa aaatttgcc gcttcctta 
      taccaagag ttggcaaaa agttgcaac aacttcgtc 
      ataagccct ttcttgatc gaaacagct ttcgctgca 
      ctttccttg ggaagctcc cgcaaaacg gctggagaa 
      ttccgcacc gtgttccac ttaccccaa acacatgaa 
      gcaattaaa ggacgatac aaacacttc ttctctctt 
      tttaccaac aaaggctac aaattccaa actgcaagc 
      aggatttcg ctcctggac gagctcgaa ctcgcccaa 
      aaatggcaa aaacaagtc cgtgatatt ttcaacatc 
      caggccgaa gtcctggac gtcaccaac aaaaatgcg 
      agtgcggaa aaaatcgat agttggttt cgggagaaa 
      acgtacaac aaagtcgat aaaattgta acacctaaa 
      gatttgaac gttgaaacg ggctttttc ctgctcagt 
      gcctcacat tttggggct tgctgggcc aaggagttt 
      gcgagcaaa cgcacacac aaggataat ttctacaac 
      agcgggaaa ttaacgcaa aagacgtcc gttgatacg 
      atgtatctt cttggatat ttcaattac ggacaaagc 
      aagcaatta gaagccaag tttttggaa ctaaatttt 
      atagaacaa caagcttcc atggtcatc attcttccg 
      gataaaatc gacgggata caaaaactt gaaaccaaa 
      atggatcaa gtgcttcag ttccgagac atgaggacc 
      acagcagtg gacatttcg ctgccaaaa ttccgaatg 
      gacagccac ttcgacctc ataaagttt ttacaaaaa 
      ttgggcata aaacaagct tttatagat ggtaaagct 
      tcctttgat gggacggta gaaactgat tgtgaccta 
      ccttacatc actaaagtc acccagaaa acttccatg 
      gaaattaac gaacgagga gtcgaagcc acagctgat 
      ccagcacat gctgcgccc ccttctgtt gcccccagt 
      tatgtgaaa tttaaagcc gaccaccca ttcatattt 
      tacatcaaa atcaaagaa gtcgtggtt tatgttggc 
      cgagtcctt gcgttttaa
    mRNA sequence
    • >TC034060 RA
      aaaaatacg aaagcacaa aattagaaa ataaatact 
      ctcgtgttg taataaaat tttaagcaa aacaatcat 
      aaagaacaa aaaaagtta atcaagcag ttgttaatc 
      caaacaatg ggaaaaacc tggcttaaa tagtaatta 
      aaacaagtt tagaacgct tattgtttt ataggactg 
      tatttaaat gaacccaaa tccatctca ttttcaaaa 
      cgatcttaa ctggggcca aaatgagta ttttcctgt 
      tttttatac tagctgtcg caattgcga caccaaaaa 
      aaccagacc ttaaaaata tacaagact atggctttt 
      ccgcaactt tgtacaagg agttgggaa aagattgtc 
      acaactttg tcataagtc ccttgttgg ttgagacag 
      ctttcgctg ccctctatt tgggaagct ctggcaaaa 
      cagctgaag aaatacgca ccgtgtttc acttacctc 
      caaaaactg gcattgaac acaagtata aaacactca 
      tcccgcagt taaccaatc ccggctata cactaaaca 
      tagcaaaca gggtttctc tagcccaaa aatacaaat 
      ttgtaccga cttgggaga cgcaaatca agaacattt 
      tcgaagtgt gtgctgatc ttttggata tgacgaata 
      aaaacgcag ctgcggaaa aagtcaata aatggtttc 
      gagacaaaa cgaacaata aaatcgaca aaatggtgt 
      ccgctgatg ctttgacaa gtgagacag ctcttttgc 
      ttttgagct cgttgtatt tccaaggca actgggatc 
      aacagtttt ctgtatcta gcacacgta aacagacgt 
      tctacaacc atggaaaac agcaagtct cagtcgata 
      tgatgtttg catcagctc ggtccgtta attacggaa 
      agagcgatc ttttaaacg ctcaattct tggaaatag 
      cgtttgaaa aacaccacg cttcgatgg ttgtagttc 
      ttccagatc aactggacg gcatttacc aacttgaaa 
      cgaagatta atgaaattt tcaattttc gcgactttt 
      gtcccgaga cggttaata tttggctgc cgaaatttc 
      gcatggata gcactttca acatcgaca agattctac 
      aacaattgg gtgtgaaag atgcctttg acgaggaaa 
      aagcccaat tcagtggaa ctgtgtcat gtgccgatt 
      gtcactcgc cgtacatcg gcaaagtta ctcaaaaaa 
      cttatctgc aaatcaatg aacgaggag ttgaagcag 
      cagctggtc aagtagtac atgttcgtg ttccgttgt 
      ctggtgggc gcaagcaat ttaaagccg accatcctt 
      tcctgtttt atatcaaga taaaagaag ttgtggtct 
      ttattggac gatttcggg agaaaacga acaataaaa 
      tcgacaaaa tggtgtccg ctgatgctt tgacaagtg 
      agacagctc ttttgcttt tgagctcgt tgtatttcc 
      aaggcaact gggatcaac agttttctg tatctagca 
      cacgtaaac agacgttct acaaccatg gaaaacagc 
      aagtctcag tcgatatga tgtttgcat cagctcggt 
      ccgttaatt acggaaaga gcgatcttt taaacgctc 
      aattcttgg aaatagcgt ttgaaaaac accacgctt 
      tgatggttg tagttcttc cagatcaac tggacggca 
      tttaccaac ttgaaacga agattagtg aaattttca 
      attttcgcg acttttgtc ccgggacgg ttaatattt 
      ggctgccga aatttcgca tggatagca ctttcaaca 
      tcgacaaga ttctacaac aattgggcg tgaaagatg 
      cctttgacg agacaaaag ctcaattca gtggaactg 
      tgtcatgtg ccgattgtc actcgccgt acatcggca 
      aagttaccc aaaaaactt atctgcaaa tcaatgaac 
      gaggagttg aagcagcag ctggtcaag tagtacatg 
      ttcgtgttc cgttgtctg gtgggcgca agcaattta 
      aagccgacc atcctttcc tgttttaca tcaagataa 
      aagaagttg tggtcttta ttggacgag ttttaatca 
      aaccagacg tgtttatta tttgctcaa tcccgggca 
      aaaaaatgc attttctct tatttttga ctttgtgct 
      tcgcatctg tatccccat acaccgaaa tacatgaca 
      aaaaatttg ccgcttcct tataccaag agttggcaa 
      aaagttgca acaacttcg tcataagcc ctttcttga 
      tcgaaacag ctttcgctg cactttcct tgggaagct 
      cccgcaaaa cggctggag aattccgca ccgtgttcc 
      acttacccc aaacacatg aagcaatta aaggacgat 
      acaaacact tcttctctc tttttacca acaaaggct 
      acaaattcc aaactgcaa gcaggattt cgctcctgg 
      acgagctcg aactcgccc aaaaatggc aaaaacaag 
      tccgtgata ttttcaaca tccaggccg aagtcctgg 
      acgtcacca acaaaaatg cgagtgcgg aaaaaatcg 
      atagttggt ttcgggaga aaacgtaca acaaagtcg 
      ataaaattg taacaccta aagatttga acgttgaaa 
      cgggctttt tcctgctca gtgcctcac attttgggg 
      cttgctggg ccaaggagt ttgcgagca aacgcacac 
      acaaggata atttctaca acagcggga aattaacgc 
      aaaagacgt ccgttgata cgatgtatc ttcttggat 
      atttcaatt acggacaaa gcaagcaat tagaagcca 
      agtttttgg aactaaatt ttatagaac aacaagctt 
      ccatggtca tcattcttc cggataaaa tcgacggga 
      tacaaaaac ttgaaacca aaatggatc aagtgcttc 
      agttccgag acatgagga ccacagcag tggacattt 
      cgctgccaa aattccgaa tggacagcc acttcgacc 
      tcataaagt ttttacaaa aattgggca taaaacaag 
      cttttatag atggtaaag cttcctttg atgggacgg 
      tagaaactg attgtgacc taccttaca tcactaaag 
      tcacccaga aaacttcca tggaaatta acgaacgag 
      gagtcgaag ccacagctg atccagcac atgctgcgc 
      ccccttctg ttgccccca gttatgtga aatttaaag 
      ccgaccacc cattcatat tttacatca aaatcaaag 
      aagtcgtgg tttatgttg gccgagtcc ttgcgtttt 
      aaataaatt gataaataa aaaattgca caatag
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