Status
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Gene information
General information
TC030889
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.

TC004741,TC004740
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_007416.3: 6284540..6295459
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
    TC030889
    iBeetle screen iB_02952

    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 embryonic musculature and early development
    number of eggs/embryos/larvae on slide: > 50 larval musculature 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
    legs shape irregular - Penetrance (percentage of animals/eggs on slide): 50% - 80%
    legs broader - Penetrance (percentage of animals/eggs on slide): 50% - 80%

    Comment: Specifically the femur, tibiotarsus and pretarsus are affected.
    affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): 50% - 80%
    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_03829

    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 embryonic musculature and early development
    number of eggs/embryos/larvae on slide: > 50 larval musculature fluorescence decreased - Penetrance (percentage of animals/eggs on slide): 30% - 50%

    affected eggs/embryos/larvae total percentage of affected eggs/embryos/larvae on slide - Penetrance (percentage of animals/eggs on slide): 30% - 50%
    Analysis of larval stage 1 cuticle
    number of eggs/embryos/larvae on slide: > 50
    legs shortened - Penetrance (percentage of animals/eggs on slide): 50% - 80%

    Comment: Tibiotarsus & Femur potentially fused.
    legs pretarsus shortened - Penetrance (percentage of animals/eggs on slide): 30% - 50%

    affected eggs/embryos/larvae total percentage of affected eggs/embryos/larvae on slide - Penetrance (percentage of animals/eggs on slide): 50% - 80%
    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: 70.0% (includes death as larva, prepupa, pupa, adult)

    eclosion delayed (still pupa) - Number of animals showing the phenotype: 5 (16 dpi)
    Days post injection


    eclosion not fulfilled (died as pupa) - Number of animals showing the phenotype: 1 (16 dpi)
    Days post injection


    adult lethality - Number of animals showing the phenotype: 1 (19 dpi)
    Days post injection


    eclosion not fulfilled (died as pupa) - Number of animals showing the phenotype: 5 (19 dpi)
    Days post injection


    Adult morphology
    elytra not closed - Number of animals showing the phenotype: 4


    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC030889 RA
      MQDENENKS LLFLNNSNK YGAMHSDSS GTPKKVIFC 
      VHGKPSAGC CKVLQSEVI DIKEGDSVD TPTKENQDA 
      TEMINKHCH RGKSPEIDR KARRKLIVA SVLCVLFMI 
      GEVIGGYLS NSLAIASDA AHLLTDFAS FMISLFALW 
      MATRPSTKT MHFGWYRAE VIGALTSVL LIWVVTGIL 
      VYIAVQRVI HKSFDIDSK VMLITSGIG VIVNLIMGL 
      TLHQHGHSH GGGSTHSHD NINVRAAFI HVIGDFLQS 
      FGVFVAAAV IYFNEQWVI IDPICTFIF SVLVMFTTF 
      NIIKDTLTV LMEGAPKGI DFNQVMKTL LNIEGVKRV 
      HNLRIWGLS LDKIAMSAH VAISPDVNP QNVLMVATK 
      NIHDKYNFF EMTLQIEEF QEAMEDFKA RLRWWIPKL 
      LFDSRDNQV GYVTSGGSI TRRSLIGWR DTPPAPKRL 
      KFRGSFFDF FTENKVKMP KDIVSCRSP YSRDLTLPD 
      LDPPPHFWT RMKQRWNGG HQLLPTTDS PTISACTTS 
      TNLSNLEFQ SSSTLNQQI YYGNNLRRA VPEIEPLQP 
      HKNNLDMTI NATQQDFEY SNESLSRSK NDKSSITTR 
      ALFRQVVAA VAVSWVSMV IGYVSGYTS PAGISLKAD 
      LQITDLQFS WISGLMPLA ALLGGLLGG PLIEGLGRK 
      WTLLLTNVL FLVSWIINY FAQEYWYLY ISRSISGCG 
      VGIASLTLP VYLGETLQP EVRGTLGLL PTAFGNIGI 
      LLCFSMGIV SEWKGIAGI GALLAVPFL FVIWFIPET 
      PRWYISKNK TDQSRRALE WLRDKNNQD TLEKEFEEL 
      LKSQKIADE KADKLKDLY SRPYVKSLL IVLGLMFFQ 
      QFSGINAVI FYTTQIFED TGSDIDSSV QTIIVGAVN 
      FASTFIATI LIDRLGRKV LLYISSVAM IITLAALGA 
      YFYLMTVPD IDIAPYSWM PLASFVVYV LGFSFGFGP 
      IPWLMMGEI LPAKIRGPA ASIATGFNW TCTFVVTTT 
      FPIFKDIIG AHGTFWLFC AVCVLGLVF TIFWVPETK 
      GQSLEDIER KLAGEKVRR MSSVANMKP LQSTF
    CDS sequence
    • >TC030889 RA
      atgcaggat gaaaacgaa aataagtct cttctattt 
      ttaaataat agtaacaaa tacggagca atgcacagt 
      gacagtagc ggcactcct aaaaaagtg atattttgt 
      gttcatgga aaaccaagt gctggttgc tgcaaagtt 
      ttacaaagt gaagtcatc gacatcaaa gagggagat 
      tcggttgat acaccgacc aaagaaaat caggatgcc 
      accgaaatg attaataaa cattgccat cgagggaaa 
      tctccggaa attgatcgc aaagctagg cgaaaatta 
      attgtagct agtgtatta tgtgtactt ttcatgatc 
      ggagaagtt ataggtggc tacctgtca aatagcttg 
      gcaatagcc tctgatgcc gctcacctt cttaccgac 
      tttgcgagt tttatgata tctctgttt gccttatgg 
      atggcaact aggccctcc accaagaca atgcacttt 
      gggtggtac cgcgctgaa gtgataggt gctctgact 
      tccgtccta ctaatttgg gtagttact ggaatactg 
      gtctacatt gccgtacaa agagtcatc cataagtcc 
      tttgatatc gatagtaaa gtaatgcta atcacctcc 
      ggtatcggc gttattgtg aacttaata atgggttta 
      acactccac caacatggg cattcccat ggaggtggt 
      agcacacat tcccacgat aacatcaac gtcagagcg 
      gcttttatt cacgttatc ggcgacttt ttgcaaagt 
      tttggtgtt ttcgtggcc gctgctgtg atttatttc 
      aacgaacaa tgggtcata attgacccc atctgtacc 
      ttcattttt tcagtatta gtgatgttt accacgttc 
      aatattatc aaagatacg ttaacagtg ctgatggaa 
      ggagctccc aaaggaatt gatttcaat caagtcatg 
      aaaacgctt ctaaatata gaaggagtt aaaagagtt 
      cataacttg agaatatgg ggcctaagt ttggataaa 
      attgcaatg tctgctcat gttgctatt agtcctgat 
      gtgaatcca caaaatgtt ttaatggtc gctactaaa 
      aatattcat gataaatac aacttcttt gagatgaca 
      ctgcagatc gaggagttc caagaggcc atggaagac 
      tttaaggca aggttaagg tggtggata cccaaattg 
      ttatttgac tcacgggac aatcaagtt ggctacgtg 
      acgtcggga ggctcgatc accaggcgc tcgctgatt 
      ggctggcgg gacacacct ccggctccg aaacgtctg 
      aaatttcgc gggtcgttt ttcgatttt tttactgaa 
      aataaagtg aaaatgcca aaagacatt gtgtcatgc 
      cggagtcct tattccagg gatttaacg ctcccagac 
      ctggaccca cccccgcac ttctggacc cgaatgaag 
      caaagatgg aacggtggt catcagttg cttccaacg 
      acagacagc ccaactatc tctgcctgt acgacttcc 
      acaaatttg tcgaatttg gaattccag tccagttcg 
      actctcaat caacaaatt tactacggg aacaactta 
      agaagagcg gtacctgaa atcgaaccc cttcaacct 
      cacaagaat aatctcgac atgacaata aatgcaacg 
      caacaagat ttcgaatat tccaatgag tctctctct 
      agatcaaaa aatgacaag tcctcgatc accaccagg 
      gcgctgttt cgccaagtt gtagcagca gttgccgtc 
      tcatgggtg tccatggtg attgggtat gtctctggt 
      tatacttca ccagctggc atttcctta aaggcagat 
      ctccaaata acagactta cagttttcg tggatctca 
      ggcctcatg ccattagcc gcccttctt ggagggttg 
      ttaggaggg cccttaatt gagggttta ggaagaaaa 
      tggactctt ttgttgacc aacgttctc ttcttagtt 
      agttggata ataaattat ttcgctcaa gaatactgg 
      tacttgtac atcagtcgc agcatctcc ggctgtggc 
      gtagggata gcttctttg acccttccg gtttacttg 
      ggagagaca ctgcagcca gaagtacgt ggcacttta 
      ggcttgttg cccacagct ttcggaaac attgggatt 
      ttgttatgt ttttcaatg ggaattgtc tcggaatgg 
      aaaggaatc gcgggaatt ggggccttg ctagctgtg 
      cctttcctc ttcgtgata tggtttata ccggaaacg 
      cccagatgg tacatttcg aagaacaaa accgaccag 
      tcacgccgg gcactggaa tggttgcga gacaagaac 
      aaccaagac acccttgag aaggagttc gaagagttg 
      ctgaaatct caaaaaatt gccgacgaa aaggccgac 
      aagcttaaa gatttgtac agtcggcct tacgtaaaa 
      tctctactg attgtgctt ggattgatg tttttccaa 
      cagttcagc ggaatcaac gctgttatc ttctacacg 
      actcaaatt tttgaagat accggttcg gacattgac 
      tccagcgtt cagaccata attgttggc gctgttaat 
      ttcgcctct acgttcatt gcaactatt ctgatcgac 
      agactgggg cgaaaagtt ctcctgtac atttcaagt 
      gttgccatg atcataact ttagcagca ctgggggcc 
      tacttctat ttaatgacc gtgccagat attgacatt 
      gcgccgtac agctggatg ccacttgcc agttttgtc 
      gtttatgtg ttgggattt tctttcggc tttgggcca 
      attccctgg ctgatgatg ggcgaaatt ttgccggcg 
      aaaattcga ggacctgca gcctccata gccacaggg 
      tttaactgg acttgcact ttcgtcgta acaacgaca 
      tttccgata ttcaaagac attattggg gcgcacggc 
      actttctgg ctgttttgc gcagtgtgt gtattagga 
      cttgttttt acaatcttc tgggtgcca gaaacgaag 
      ggacagagt ttagaggac attgaaaga aaattagcg 
      ggtgaaaaa gtgcgaaga atgagttcc gtagcgaac 
      atgaagcca ttgcaatct acattttag
    mRNA sequence
    • >TC030889 RA
      tcttgtttg tatttattc aactaggta cctacttga 
      aatagataa gaaactgtc gggccgggt gcacaaaca 
      ctccttagt tcacaaaga taacaataa cttattgcc 
      cagaaactg aacatgaac atattctga cattctgag 
      attaatccg tgcccacgt aaatgaaaa acaaactgg 
      caagtggtt gaatagcca taaaaatgc aggatgaaa 
      acgaaaata agtctcttc tatttttaa ataatagta 
      acaaatacg gagcaatgc acagtgaca gtagcggca 
      ctcctaaaa aagtgatat tttgtgttc atggaaaac 
      caagtgctg gttgctgca aagttttac aaagtgaag 
      tcatcgaca tcaaagagg gagattcgg ttgatacac 
      cgaccaaag aaaatcagg atgccaccg aaatgatta 
      ataaacatt gccatcgag ggaaatctc cggaaattg 
      atcgcaaag ctaggcgaa aattaattg tagctagtg 
      tattatgtg tacttttca tgatcggag aagttatag 
      gtggctacc tgtcaaata gcttggcaa tagcctctg 
      atgccgctc accttctta ccgactttg cgagtttta 
      tgatatctc tgtttgcct tatggatgg caactaggc 
      cctccacca agacaatgc actttgggt ggtaccgcg 
      ctgaagtga taggtgctc tgacttccg tcctactaa 
      tttgggtag ttactggaa tactggtct acattgccg 
      tacaaagag tcatccata agtcctttg atatcgata 
      gtaaagtaa tgctaatca cctccggta tcggcgtta 
      ttgtgaact taataatgg gtttaacac tccaccaac 
      atgggcatt cccatggag gtggtagca cacattccc 
      acgataaca tcaacgtca gagcggctt ttattcacg 
      ttatcggcg actttttgc aaagttttg gtgttttcg 
      tggccgctg ctgtgattt atttcaacg aacaatggg 
      tcataattg accccatct gtaccttca ttttttcag 
      tattagtga tgtttacca cgttcaata ttatcaaag 
      atacgttaa cagtgctga tggaaggag ctcccaaag 
      gaattgatt tcaatcaag tcatgaaaa cgcttctaa 
      atatagaag gagttaaaa gagttcata acttgagaa 
      tatggggcc taagtttgg ataaaattg caatgtctg 
      ctcatgttg ctattagtc ctgatgtga atccacaaa 
      atgttttaa tggtcgcta ctaaaaata ttcatgata 
      aatacaact tctttgaga tgacactgc agatcgagg 
      agttccaag aggccatgg aagacttta aggcaaggt 
      taaggtggt ggataccca aattgttat ttgactcac 
      gggacaatc aagttggct acgtgacgt cgggaggct 
      cgatcacca ggcgctcgc tgattggct ggcgggaca 
      cacctccgg ctccgaaac gtctgaaat ttcgcgggt 
      cgtttttcg attttttta ctgaaaata aagtgaaaa 
      tgccaaaag acattgtgt catgccgga gtccttatt 
      ccagggatt taacgctcc cagacctgg acccacccc 
      cgcacttct ggacccgaa tgaagcaaa gatggaacg 
      gtggtcatc agttgcttc caacgacag acagcccaa 
      ctatctctg cctgtacga cttccacaa atttgtcga 
      atttggaat tccagtcca gttcgactc tcaatcaac 
      aaatttact acgggaaca acttaagaa gagcggtac 
      ctgaaatcg aaccccttc aacctcaca agaataatc 
      tcgacatga caataaatg caacgcaac aagatttcg 
      aatattcca atgagtctc tctctagat caaaaaatg 
      acaagtcct cgatcacca ccagggcgc tgtttcgcc 
      aagttgtag cagcagttg ccgtctcat gggtgtcca 
      tggtgattg ggtatgtct ctggttata cttcaccag 
      ctggcattt ccttaaagg cagatctcc aaataacag 
      acttacagt tttcgtgga tctcaggcc tcatgccat 
      tagccgccc ttcttggag ggttgttag gagggccct 
      taattgagg gtttaggaa gaaaatgga ctcttttgt 
      tgaccaacg ttctcttct tagttagtt ggataataa 
      attatttcg ctcaagaat actggtact tgtacatca 
      gtcgcagca tctccggct gtggcgtag ggatagctt 
      ctttgaccc ttccggttt acttgggag agacactgc 
      agccagaag tacgtggca ctttaggct tgttgccca 
      cagctttcg gaaacattg ggattttgt tatgttttt 
      caatgggaa ttgtctcgg aatggaaag gaatcgcgg 
      gaattgggg ccttgctag ctgtgcctt tcctcttcg 
      tgatatggt ttataccgg aaacgccca gatggtaca 
      tttcgaaga acaaaaccg accagtcac gccgggcac 
      tggaatggt tgcgagaca agaacaacc aagacaccc 
      ttgagaagg agttcgaag agttgctga aatctcaaa 
      aaattgccg acgaaaagg ccgacaagc ttaaagatt 
      tgtacagtc ggccttacg taaaatctc tactgattg 
      tgcttggat tgatgtttt tccaacagt tcagcggaa 
      tcaacgctg ttatcttct acacgactc aaatttttg 
      aagataccg gttcggaca ttgactcca gcgttcaga 
      ccataattg ttggcgctg ttaatttcg cctctacgt 
      tcattgcaa ctattctga tcgacagac tggggcgaa 
      aagttctcc tgtacattt caagtgttg ccatgatca 
      taactttag cagcactgg gggcctact tctatttaa 
      tgaccgtgc cagatattg acattgcgc cgtacagct 
      ggatgccac ttgccagtt ttgtcgttt atgtgttgg 
      gattttctt tcggctttg ggccaattc cctggctga 
      tgatgggcg aaattttgc cggcgaaaa ttcgaggac 
      ctgcagcct ccatagcca cagggttta actggactt 
      gcactttcg tcgtaacaa cgacatttc cgatattca 
      aagacatta ttggggcgc acggcactt tctggctgt 
      tttgcgcag tgtgtgtat taggacttg tttttacaa 
      tcttctggg tgccagaaa cgaagggac agagtttag 
      aggacattg aaagaaaat tagcgggtg aaaaagtgc 
      gaagaatga gttccgtag cgaacatga agccattgc 
      aatctacat tttagatta aatgactga tttttttat 
      ttagacgta agatttgta gatctgtat atacattaa 
      attattttt atatgtaaa taaattatt atgttttat
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