Status
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Gene information
General information
TC032431
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.

TC003608,TC003607
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_007418.3: 8852430..8868590
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
    TC032431
    iBeetle screen iB_00589

    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


    Fertility
    mature egg sometimes not present - Number of animals showing the phenotype: 4

    tropharium irregular - Number of animals showing the phenotype: 4
    vitellogenic egg chamber number decreased - Number of animals showing the phenotype: 4
    Analysis of embryonic musculature and early development
    number of eggs/embryos/larvae on slide: < 50 embryo/egg with embryonic tissue, no cuticle, but muscle fluorescence - Penetrance (percentage of animals/eggs on slide): 50% - 80%

    embryo/egg not developed/not fertilized - Penetrance (percentage of animals/eggs on slide): 50% - 80%

    affected eggs/embryos/larvae total percentage of affected eggs/embryos/larvae on slide - Penetrance (percentage of animals/eggs on slide): more than 80%
    Analysis of larval stage 1 cuticle
    number of eggs/embryos/larvae on slide: > 50
    egg without cuticle embryo/egg without visible larval cuticle in cuticle preparation - Penetrance (percentage of animals/eggs on slide): 30% - 50%

    cuticle remnants with unidentifyable segments - Penetrance (percentage of animals/eggs on slide): 30% - 50%

    Comment: cuticle remnants upended (inside-outed)
    larva everted (inside-out) - 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): more than 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: 20.0% (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: 2 (11 dpi)
    Days post injection


    pupal molt delayed (still prepupa) - Number of animals showing the phenotype: 8 (11 dpi)
    Days post injection


    pupal molt not fulfilled (died as prepupa) - Number of animals showing the phenotype: 8 (16 dpi)
    Days post injection


    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC032431 RA
      MFGKPDPQQ QWSVHAEFT ISHPRSDPD NLPQAHTEQ 
      ADNGVRMTY TWTSSSQAA SAWSAPPST SSEGAPKYG 
      FQKGNLLFT STPPRRVHD SGYGSDLLS PNSYQRRPV 
      QPYNRKCRS TCNIVLSTN IDENRTEET DSVHTQCGR 
      TQSLRCQTP TLRCDRKDN FYGCGDPWC HHYEDFSGR 
      FSPVMETCE DCPSGQAST KECDNRRDA CVQTFEMID 
      KCTSPIVKM GRVKYAEYE DRKEKREKK KYGSGSRRR 
      TEPIFQRSQ SPSSITPDS LEGTKGQRR LVRKNATVG 
      KDKDSSTDS KLSDSSKKP RTVHIDVYC TGTEMESDS 
      SSESETAST PQTVFESEK VRITHKRAD ETEMPFSVR 
      KSRSKSKEK EAKRIEESD DDGASTAYP SKLSSYSTI 
      RDFESSISS VPQSWTPYS MSSCTVPDD YDSVANTSW 
      KDTFSDIDS LIHSRSSIA QTESLDFVP RKLFEKNES 
      IDETPEILE TPSQESLQP SDSFEYANS EDRLRIKKM 
      EEFWKNKTH SKHWKSPHV ERRYLLQQK KIKEYLEKR 
      LSDKNLPKG ESKDSDSDA SDGSEKGWT IVKEKNTEL 
      APPKISDIS CKSPSVLAI QQKLTMDPT LRAPFTIVP 
      GIYTNQRAI AKKFGSIVP VVRKPGHHI GPVKNPDCL 
      CEHCQRHFA SVGHRGRTR SMGDPPSSF QNWKNIGYP 
      QAESASNKR ALKTWRISV EAQDSLRAS SMKLDLLRK 
      SLEMRRAEL PADSPVAMQ LKEELANAQ TFSPLSVHY 
      TSLQPFRER SNGNKMLTS SSTINRCAA VTGTLEVRL 
      MGCQDLLED VPGRPRKEA DGSSSPMDL RFFKKGVTG 
      RGSSKSYSI KDEVSDEIM AVLKLDNTT VAQTSWKPC 
      SQQAWDQRF SIELDKHRE LEIGVYWRD WRSLCAVKF 
      LKLEEFIDD DRHGMALHL EPQGLLFAE IKFLNPMIS 
      KQPKLQRQK RIFKQVMPR AKQMNINIV TWGRWMKQS 
      SRIPNRNNT IVQSTEMLP EEPEDKPET PGETPDPQA 
      FGLGGQRPL GLGVAVLPE TPPPVPTAP PPIAKKRMC 
      VTPPPVPPR LDPESAAAL KEFDFLEEQ EMSRTGPLL 
      IPSTPSPVV IPEQLVPPS PQPVIEFPD DEPPLEAVN 
      VARRPVARE LGYRDSAYE SRRHSQYTG MSIDNFKLI 
      SVLGRGHFG KVILSQYRN TGEYFAIKA LKKGDIIAR 
      DEVESLLSE KRIFEVANS IRHPFLVNL FACFQTEAH 
      VCFVMEYAA GGDLMMHIH ADVFSEPRA VFYAACVVL 
      GLQYLHENK IIYRDLKLD NLLLDTEGY VKIADFGLC 
      KEGMGFGDR TGTFCGTPE FLAPEVLTE TSYTRAVDW 
      WGLGVLIFE MLVGESPFP GDDEEEVFD SIVNDEVRY 
      PRFLSLESI AIMRRLLRK IPDRRLGSS ERDAEDVKK 
      QAFFRHIQW DELLHRRVP PPFVPTVHS MEDVSNFDE 
      EFTSEKPHL TPPKEPRPL TEQDQYLFR DFTYIADWC
    CDS sequence
    • >TC032431 RA
      atgtttgga aaaccggac ccccagcag caatggtcc 
      gtccacgcg gagttcacc atctcccat cctaggtcc 
      gaccccgac aacctgccc caggcccac accgagcag 
      gccgacaac ggcgtgcgc atgacctac acctggacc 
      tcctcctcg caggcggcg agcgcctgg agcgccccg 
      ccgtccacg tccagcgag ggcgccccc aagtacggc 
      ttccagaag ggcaatctg ctcttcacg tccacgccg 
      ccgaggagg gtccacgat tcgggctac ggaagcgac 
      ctgctgtcg ccgaactcg taccaaaga cggccggtc 
      cagccgtac aatcgcaaa tgtcgcagc acctgcaac 
      attgtcctt tcgaccaac atcgatgaa aatagaacg 
      gaagagacg gactcggtt catacgcaa tgcggaaga 
      acgcagagt ttgcgttgc caaacgccc acgttgcgg 
      tgcgaccga aaggataat ttttacgga tgtggcgac 
      ccttggtgc caccactac gaggacttc tccgggagg 
      ttctcgccg gtgatggag acgtgcgag gactgtcct 
      agtgggcaa gcctcgacg aaggaatgc gataatcgc 
      agggatgcg tgtgtgcaa acgttcgaa atgattgat 
      aagtgtacc tcgccgatt gttaaaatg gggagggtg 
      aagtatgcg gagtacgag gatcggaag gagaagagg 
      gagaagaag aagtacggg tcagggagt cggaggcgg 
      acagaaccg atttttcaa aggtcacaa tcaccgtcc 
      tcaatcacg ccagatagt ctagagggg acgaagggt 
      cagcggcgc ctggtgcgt aagaatgca acagttggc 
      aaagataaa gactcatcg actgattca aagttaagc 
      gattcgtcg aaaaaaccg cgaactgtt cacattgac 
      gtctactgt accggaacc gaaatggaa tcggattct 
      tcatcggaa tcagagaca gcgtcaacg ccacaaacg 
      gtgttcgaa tcggaaaaa gtgcgaata acgcacaaa 
      cgtgccgat gaaactgaa atgccattc agtgttcgc 
      aaatcacga tcgaaaagt aaagagaaa gaggcgaag 
      cgaatcgaa gagtccgac gatgacggt gctagtacg 
      gcctatccg tcgaaattg agctcatat tcaacgata 
      agggatttc gagtctagt atttccagt gtccctcag 
      tcctggacc ccgtattcc atgtccagt tgtacggtt 
      ccggatgat tacgattcg gtggcaaac acttcgtgg 
      aaagacact ttttccgat attgatagt ctgattcat 
      agtcgttcc agtattgct cagaccgaa agtctggat 
      tttgttcct agaaaattg ttcgaaaag aatgaaagt 
      atagatgag acacccgaa attttggaa actccaagt 
      caagagagt ctccaacct agtgatagt ttcgaatat 
      gccaattca gaggatcgt ctcaggatc aagaaaatg 
      gaagaattt tggaagaac aagactcat tccaaacac 
      tggaaatcg ccacatgtg gaacgccgg tacttgctg 
      caacagaag aagattaag gagtacttg gagaaacgc 
      ttgagtgat aagaattta ccaaaaggc gagtcgaaa 
      gactcagac tctgacgct agcgacggt tcggagaaa 
      ggctggacg atagtgaag gaaaaaaac accgaatta 
      gcgccgccg aaaatcagt gatatttcg tgtaaatcc 
      cctagtgtt ttggccatc cagcaaaaa ttgaccatg 
      gacccgaca ctacgcgct cctttcacc atagttccc 
      ggcatttat accaatcaa cgagctatc gcgaaaaaa 
      ttcggctcg attgtaccc gtagtgcgt aagccgggc 
      catcatatc ggtccggtt aaaaacccc gactgtctt 
      tgcgaacac tgtcaaaga cactttgcg agtgttggg 
      cacaggggg cgcacgcgg tccatgggc gacccgcca 
      tcatcgttt caaaactgg aaaaacatt ggttatccg 
      caagctgaa tcggcgtca aataaaaga gctcttaag 
      acgtggaga ataagcgtc gaggcacag gacagtttg 
      agggccagc agtatgaag ttggatttg ttgcggaag 
      agtttggag atgcggcgg gcggaattg cccgccgat 
      tcgcccgtc gcgatgcaa ttgaaggaa gaactggca 
      aatgctcaa acatttagt cccttatct gttcactat 
      acgagtttg caaccgttt cgggagcga tcaaacggg 
      aataaaatg ttgacgtcg tcgtcaaca atcaaccga 
      tgtgcagcc gtcacaggt accttggaa gtgagactc 
      atgggttgt caggatttg ctcgaggat gtgccagga 
      agaccgaga aaggaggcc gatgggagc tcgagtccc 
      atggatttg aggtttttc aagaaagga gtgacggga 
      agagggtcg tcgaagagt tacagtatt aaagatgaa 
      gttagtgac gagataatg gcagtgctc aagcttgac 
      aatacgaca gttgctcaa accagttgg aaaccgtgc 
      tcacaacaa gcctgggat caaagattc tcaatagaa 
      ctagacaag catcgcgag ctagaaatc ggagtttat 
      tggagagac tggaggtct ttgtgtgcc gtaaaattt 
      ttaaaactg gaggagttt attgacgac gaccgacac 
      ggaatggcg ttgcattta gaaccgcag ggtttacta 
      ttcgctgaa ataaaattc ctcaatccg atgatatcg 
      aaacagccg aaactgcaa agacagaaa cgaatattc 
      aagcaggtg atgccgcga gccaagcaa atgaacatc 
      aacatcgtc acgtggggc cgttggatg aaacaatcg 
      tcgcgaatt cccaatcgg aacaacacc attgtgcaa 
      tcgacggaa atgttgccc gaagaaccg gaggacaaa 
      ccggaaact ccgggcgaa acgccggac cctcaggcg 
      ttcggtttg ggtggccag cgtccgctc ggcttaggt 
      gtggcggtt ctgccggaa acgccgccc ccagttcct 
      acagcgccg cctccgatt gccaaaaag cggatgtgt 
      gtgacgccg ccccccgta ccgccaaga ctggatcct 
      gagagtgcc gctgcactg aaggagttt gatttctta 
      gaggaacag gagatgtct cgtacggga cctcttctg 
      attccttct acaccttca cctgtagtg attcccgag 
      caattggtt cctcctagt cctcagccc gtaattgag 
      ttcccagac gacgagccc cctctggag gccgtgaac 
      gtggcccgc cgcccggtg gcccgcgag ctgggctac 
      cgcgacagc gcctacgaa tcgcgccgc cactcgcaa 
      tacaccgga atgagtatc gacaacttc aaactgatc 
      agcgtctta ggtcgcggc cacttcggc aaagtcatc 
      ctctcgcag taccgcaac accggcgag tacttcgcc 
      atcaaagcc ctcaaaaag ggcgacatc atcgcccgc 
      gacgaggtc gaatccctc ctctccgag aaacgcatc 
      ttcgaagtg gccaactcc atccgacac cccttcctt 
      gtcaacctc ttcgcctgt ttccaaacc gaagcgcac 
      gtctgtttc gtcatggag tacgccgca ggaggcgac 
      ctgatgatg cacatccat gcggatgtc ttcagcgag 
      ccccgagcg gtcttttac gccgcttgt gtggtgctg 
      gggctgcag tatttgcac gagaataag atcatttat 
      cgcgatttg aagctggat aatctcctg ctggacacg 
      gagggttac gtgaagatt gccgatttc gggctctgt 
      aaggaaggg atgggattc ggggatagg acggggacg 
      ttttgcggg acgcccgag tttctggcg ccggaggtg 
      ttgacggag acgtcgtat acgcgagcg gtggattgg 
      tgggggttg ggggttttg atttttgag atgttggta 
      ggggagtca ccgttcccg ggggatgat gaggaggaa 
      gttttcgat tcgattgtt aatgatgaa gttcgttat 
      ccgaggttc ctatcgtta gagtcaata gcgatcatg 
      aggagattg ttgcgcaag attccagac cgacggtta 
      ggttccagc gaaagggat gccgaagac gttaagaaa 
      caagccttc tttagacac attcaatgg gacgagttg 
      ttgcatcgg agggttccg cccccgttc gtacccact 
      gtgcattcg atggaagac gtcagcaac tttgatgag 
      gagttcacc tccgagaag ccacacttg acgccgcca 
      aaagagccg cggcccttg accgaacaa gaccaatac 
      ctgtttagg gacttcacc tacatagcc gactggtgc 
      tga
    mRNA sequence
    • >TC032431 RA
      cacgtcgag tgcatgaaa gctcgtctc tccgaagca 
      agaagatca tcgaagtct aagaataag tttgtttac 
      gaagcgtag tgatttgga gtgcattcc ccgatcctc 
      gtccagttg cgatccgcc atcggccaa taattgcgc 
      cttatcccg gtgccacca ccctctaaa tatagccta 
      gtgcgtgcg tgacgtaac agcatgtga acggtcatg 
      tttggaaaa ccggacccc cagcagcaa tggtccgtc 
      cacgcggag ttcaccatc tcccatcct aggtccgac 
      cccgacaac ctgccccag gcccacacc gagcaggcc 
      gacaacggc gtgcgcatg acctacacc tggacctcc 
      tcctcgcag gcggcgagc gcctggagc gccccgccg 
      tccacgtcc agcgagggc gcccccaag tacggcttc 
      cagaagggc aatctgctc ttcacgtcc acgccgccg 
      aggagggtc cacgattcg ggctacgga agcgacctg 
      ctgtcgccg aactcgtac caaagacgg ccggtccag 
      ccgtacaat cgcaaatgt cgcagcacc tgcaacatt 
      gtcctttcg accaacatc gatgaaaat agaacggaa 
      gagacggac tcggttcat acgcaatgc ggaagaacg 
      cagagtttg cgttgccaa acgcccacg ttgcggtgc 
      gaccgaaag gataatttt tacggatgt ggcgaccct 
      tggtgccac cactacgag gacttctcc gggaggttc 
      tcgccggtg atggagacg tgcgaggac tgtcctagt 
      gggcaagcc tcgacgaag gaatgcgat aatcgcagg 
      gatgcgtgt gtgcaaacg ttcgaaatg attgataag 
      tgtacctcg ccgattgtt aaaatgggg agggtgaag 
      tatgcggag tacgaggat cggaaggag aagagggag 
      aagaagaag tacgggtca gggagtcgg aggcggaca 
      gaaccgatt tttcaaagg tcacaatca ccgtcctca 
      atcacgcca gatagtcta gaggggacg aagggtcag 
      cggcgcctg gtgcgtaag aatgcaaca gttggcaaa 
      gataaagac tcatcgact gattcaaag ttaagcgat 
      tcgtcgaaa aaaccgcga actgttcac attgacgtc 
      tactgtacc ggaaccgaa atggaatcg gattcttca 
      tcggaatca gagacagcg tcaacgcca caaacggtg 
      ttcgaatcg gaaaaagtg cgaataacg cacaaacgt 
      gccgatgaa actgaaatg ccattcagt gttcgcaaa 
      tcacgatcg aaaagtaaa gagaaagag gcgaagcga 
      atcgaagag tccgacgat gacggtgct agtacggcc 
      tatccgtcg aaattgagc tcatattca acgataagg 
      gatttcgag tctagtatt tccagtgtc cctcagtcc 
      tggaccccg tattccatg tccagttgt acggttccg 
      gatgattac gattcggtg gcaaacact tcgtggaaa 
      gacactttt tccgatatt gatagtctg attcatagt 
      cgttccagt attgctcag accgaaagt ctggatttt 
      gttcctaga aaattgttc gaaaagaat gaaagtata 
      gatgagaca cccgaaatt ttggaaact ccaagtcaa 
      gagagtctc caacctagt gatagtttc gaatatgcc 
      aattcagag gatcgtctc aggatcaag aaaatggaa 
      gaattttgg aagaacaag actcattcc aaacactgg 
      aaatcgcca catgtggaa cgccggtac ttgctgcaa 
      cagaagaag attaaggag tacttggag aaacgcttg 
      agtgataag aatttacca aaaggcgag tcgaaagac 
      tcagactct gacgctagc gacggttcg gagaaaggc 
      tggacgata gtgaaggaa aaaaacacc gaattagcg 
      ccgccgaaa atcagtgat atttcgtgt aaatcccct 
      agtgttttg gccatccag caaaaattg accatggac 
      ccgacacta cgcgctcct ttcaccata gttcccggc 
      atttatacc aatcaacga gctatcgcg aaaaaattc 
      ggctcgatt gtacccgta gtgcgtaag ccgggccat 
      catatcggt ccggttaaa aaccccgac tgtctttgc 
      gaacactgt caaagacac tttgcgagt gttgggcac 
      agggggcgc acgcggtcc atgggcgac ccgccatca 
      tcgtttcaa aactggaaa aacattggt tatccgcaa 
      gctgaatcg gcgtcaaat aaaagagct cttaagacg 
      tggagaata agcgtcgag gcacaggac agtttgagg 
      gccagcagt atgaagttg gatttgttg cggaagagt 
      ttggagatg cggcgggcg gaattgccc gccgattcg 
      cccgtcgcg atgcaattg aaggaagaa ctggcaaat 
      gctcaaaca tttagtccc ttatctgtt cactatacg 
      agtttgcaa ccgtttcgg gagcgatca aacgggaat 
      aaaatgttg acgtcgtcg tcaacaatc aaccgatgt 
      gcagccgtc acaggtacc ttggaagtg agactcatg 
      ggttgtcag gatttgctc gaggatgtg ccaggaaga 
      ccgagaaag gaggccgat gggagctcg agtcccatg 
      gatttgagg tttttcaag aaaggagtg acgggaaga 
      gggtcgtcg aagagttac agtattaaa gatgaagtt 
      agtgacgag ataatggca gtgctcaag cttgacaat 
      acgacagtt gctcaaacc agttggaaa ccgtgctca 
      caacaagcc tgggatcaa agattctca atagaacta 
      gacaagcat cgcgagcta gaaatcgga gtttattgg 
      agagactgg aggtctttg tgtgccgta aaattttta 
      aaactggag gagtttatt gacgacgac cgacacgga 
      atggcgttg catttagaa ccgcagggt ttactattc 
      gctgaaata aaattcctc aatccgatg atatcgaaa 
      cagccgaaa ctgcaaaga cagaaacga atattcaag 
      caggtgatg ccgcgagcc aagcaaatg aacatcaac 
      atcgtcacg tggggccgt tggatgaaa caatcgtcg 
      cgaattccc aatcggaac aacaccatt gtgcaatcg 
      acggaaatg ttgcccgaa gaaccggag gacaaaccg 
      gaaactccg ggcgaaacg ccggaccct caggcgttc 
      ggtttgggt ggccagcgt ccgctcggc ttaggtgtg 
      gcggttctg ccggaaacg ccgccccca gttcctaca 
      gcgccgcct ccgattgcc aaaaagcgg atgtgtgtg 
      acgccgccc cccgtaccg ccaagactg gatcctgag 
      agtgccgct gcactgaag gagtttgat ttcttagag 
      gaacaggag atgtctcgt acgggacct cttctgatt 
      ccttctaca ccttcacct gtagtgatt cccgagcaa 
      ttggttcct cctagtcct cagcccgta attgagttc 
      ccagacgac gagccccct ctggaggcc gtgaacgtg 
      gcccgccgc ccggtggcc cgcgagctg ggctaccgc 
      gacagcgcc tacgaatcg cgccgccac tcgcaatac 
      accggaatg agtatcgac aacttcaaa ctgatcagc 
      gtcttaggt cgcggccac ttcggcaaa gtcatcctc 
      tcgcagtac cgcaacacc ggcgagtac ttcgccatc 
      aaagccctc aaaaagggc gacatcatc gcccgcgac 
      gaggtcgaa tccctcctc tccgagaaa cgcatcttc 
      gaagtggcc aactccatc cgacacccc ttccttgtc 
      aacctcttc gcctgtttc caaaccgaa gcgcacgtc 
      tgtttcgtc atggagtac gccgcagga ggcgacctg 
      atgatgcac atccatgcg gatgtcttc agcgagccc 
      cgagcggtc ttttacgcc gcttgtgtg gtgctgggg 
      ctgcagtat ttgcacgag aataagatc atttatcgc 
      gatttgaag ctggataat ctcctgctg gacacggag 
      ggttacgtg aagattgcc gatttcggg ctctgtaag 
      gaagggatg ggattcggg gataggacg gggacgttt 
      tgcgggacg cccgagttt ctggcgccg gaggtgttg 
      acggagacg tcgtatacg cgagcggtg gattggtgg 
      gggttgggg gttttgatt tttgagatg ttggtaggg 
      gagtcaccg ttcccgggg gatgatgag gaggaagtt 
      ttcgattcg attgttaat gatgaagtt cgttatccg 
      aggttccta tcgttagag tcaatagcg atcatgagg 
      agattgttg cgcaagatt ccagaccga cggttaggt 
      tccagcgaa agggatgcc gaagacgtt aagaaacaa 
      gccttcttt agacacatt caatgggac gagttgttg 
      catcggagg gttccgccc ccgttcgta cccactgtg 
      cattcgatg gaagacgtc agcaacttt gatgaggag 
      ttcacctcc gagaagcca cacttgacg ccgccaaaa 
      gagccgcgg cccttgacc gaacaagac caatacctg 
      tttagggac ttcacctac atagccgac tggtgctga 
      cattagctt taggccgat ttgatgaac aacgtgctg 
      ctttaattt caactacat gaacaaatt tatgtatat 
      aagatctca gtattgaaa tcacgattc atgcaatta 
      ttgtgtaac atatcattt gtcatggtt aatttgata 
      agtcgctcg cgacggatt tttgtaatt tagtttatt 
      ggcagttct gtatatatt ttattcata ctcttgtgg 
      taaatttac tcaaagtgt tgcattttt ccccgagtt 
      ggagtcaag tcgctcttc cagtatgta tcaaagact 
      gttgcaatt cgagaatag ctgaacttg gaagtgcct 
      gtttttaaa agatcacgc tcagtgtac atagttact 
      aatcctagt gtaatctat tattttttg atgctttaa 
      atatatacc tatataaat tatgtaagt tttaataca 
      aacatgttt aagctgact tcagtaaat aatatttta 
      tttatattc tttacatca ctaaatatt tacaacgtt 
      atgaataaa gtcgtgttt atctaaata atttaattc 
      gtactttcc cttaacgtg acaggaatt ccacacagg 
      gcggcttcc gctatcacg ccccgattt ttacttatt 
      tccgcttcc agtttttcc gtcgagtgt ttgttggcg 
      cttacatca gccggcatt tgcaggcct tcgcccaac 
      ttgtaaaaa acacaagtt gagtcagcg ctgactaat 
      cctaaataa atgctgctc ttctat
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