Status
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Gene information
General information
TC015859 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_007421.3: 9695734..9716518
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
    TC015859
    iBeetle screen iB_05916

    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 embryonic musculature and early development
    number of eggs/embryos/larvae on slide: > 50 embryo/egg not developed/not fertilized - Penetrance (percentage of animals/eggs on slide): 30% - 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): 30% - 50%
    remarks: muscle pattern potentially obscured by segmentation defect,
    Analysis of larval stage 1 cuticle
    number of eggs/embryos/larvae on slide: < 20
    embryo/egg without visible larval cuticle in cuticle preparation - Penetrance (percentage of animals/eggs on slide): 50% - 80%
    head size decreased - Penetrance (percentage of animals/eggs on slide): less than 30%
    antenna not present - Penetrance (percentage of animals/eggs on slide): less than 30%

    mandible not present - Penetrance (percentage of animals/eggs on slide): less than 30%
    head number decreased - Penetrance (percentage of animals/eggs on slide): less than 30%
    thorax segment number decreased -

    thorax number decreased - Penetrance (percentage of animals/eggs on slide): less than 30%
    abdomen segment number decreased - Penetrance (percentage of animals/eggs on slide): less than 30%

    abdomen number decreased - Penetrance (percentage of animals/eggs on slide): less than 30%
    affected eggs/embryos/larvae - 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: % (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
    • >TC015859 RA
      MGETRGSIY FAQLKAMLK RNILLKKRE KRKTTAEVL 
      LPLYSLGVL IVMKVMIPN PNFPAIDTP RGEVHLFSY 
      FQKLKEHTV AVVPNTNET QDFLKRVND LWHTIDHNT 
      SGFHPIKWL QFPTEEDLL TAYWSQPDS IPIAVIFEE 
      PRPITGPLK YKIRSNPSE FETPSTTVL YSSPASCRE 
      SSDFWSGAN VLPIETGNS CPVNRYYYS GFVALQALL 
      DYTKIKIDT DRDFHVPKI SLEMFPKHA HTGGWMVAF 
      RLVIPLYMV MALSQFITY LLILIVGEK ENKIKEGMK 
      IMGLKDSVF WLSWFIIYG IFVLFLSIV CCVLLFTLQ 
      VFQNTNFLL IFLLVLLYT LSIIMFGFM ITPFFDKSR 
      TAGILGNFA VNIMSLFYF IQVFIDSSS SVAFWVVSL 
      ISSSGFALA MDKALVMEL KGEGVNFDN LWSGPGMPF 
      GGSLIMMAL DIVLYGLLA YYLDCVIPS EHGIRRSPF 
      FCFKPFFWF NKKPVQRIP LANGGSAGS LTTAEDGHN 
      DVEPVSREM KGREAIRIV DMFKTFHHC RKPEIKAIN 
      GINLTIYEG QITAILGHN GAGKTTLFN ILTGLTAPT 
      SGTAYIFGY DVRDPNDMD EIRRMTGVC PQHDILFDN 
      LTPKEHLEF FAAVKGIPP NLREFEVMK TLRDIDLTD 
      KANASAKHL SGGQKRKLS IGIAVIGDP KIIILDEPT 
      AGVDPYSRR HMWSVLQNR RHGKVILLT THFMDEADI 
      LADRKAVVS KGNIRCCGS SLFLKNKFG IGYHLTLVL 
      DGICREHAI TRLVTSHVP KAEKARRHG RELSFILPH 
      NAVDNFASL FSAIEQEIN NKSSKLGIS SYGVSMTTL 
      EEVFLHLER DEETEGTMD NLSKKMVRN RALSRSLSL 
      QSKSTSYQS LQNEGNNAT NQDGKGGGD TAQIGGLEI 
      ISETKSPIT GIGLEKIEC HPNVFQTLI SLLRLRILR 
      MIRDIQKLY FMVLLPLGL AALVLYFNS MQTIEVKLT 
      PLALNGSTY ADKTTFAIH NASGVVLDD FYDQLLEVG 
      VQRIDPYDG NFSLLLDVA PHMAALNVN VFTNDLSIT 
      VIYNDTAQH SLPIILNLI SNGLYRLMS PERVIRGQW 
      EPIEVLTLP FQQTAQPEE FDIGIFSAS TFIGMIFVL 
      VPVSLAIDM VYDREIKAK NQLRVNGLS FCMYFVTYF 
      VVLGGLMIL ICAALVLII LLFNIPAFK GWPALVTLG 
      TLTLLYCPS SILFSTCVS YIFDKTDSA QSILPNIAT 
      FVGCIPFFL VTALDMLNI GGKAAFVLH ILFSLLNAM 
      YIPYAIIYY IQRVYIMCK LNSTCNTLT LTDYMTSEI 
      IVMLVGIIV NIPFWFFVL MIIDIKKSG GRISDAFKF 
      FKRNTDENT EDINELSDI GENEDQDVK AERQKVKNL 
      MDCHVVNPP VVMVQNLHK EYTKSEIKC ECCCKHDDE 
      PDTTKIAIR SLSLAVDAG EVFGLLGHN GAGKTTTMK 
      IITAEEAPT RGRVQIGGE NITSNMNNA FQLLGYCPQ 
      HDALWKNIT VREHLECYA AIRGVPSGD IPQIVDLYL 
      TGLQIHEHA DKQAQQCSG GTRRKLSFA MAMVGNPKV 
      VLLDEPSTG MDPRSKRFL WDTILASFQ GSRGAILTT 
      HSMEEADAL CSRVGIMVK GELRCLGST QHLKNLYGA 
      GYTLEMKLR GGDRTPTST SCDRPNELR EFVTGLFHD 
      ATLQESFAD RLVFSVPQQ SVPSLANCF MQLEKVKTE 
      LDIEEYSFS QTTLEQVFL KFAQYDEVN GE
    CDS sequence
    • >TC015859 RA
      atgggggag acgcgaggc tccatctac tttgctcag 
      ctgaaggcc atgctgaag cggaatatt ctgctgaag 
      aagagggaa aagaggaaa acgactgcg gaggtcttg 
      ttgcctttg tactcgctt ggcgtcctc atcgtgatg 
      aaagtaatg attccaaac ccaaatttc cccgcaata 
      gacacgcca cgaggtgaa gtccatctc ttctcctat 
      ttccaaaaa ctgaaagaa cacaccgtc gccgtagtt 
      cccaacact aacgaaaca caagacttt ttaaaacgc 
      gtcaacgac ttgtggcac accatcgac cacaacacg 
      tcaggtttt catccaatc aaatggcta cagttccca 
      accgaagag gatctactt actgcatat tggtcacaa 
      ccggacagt ataccaatc gctgtgatt ttcgaagaa 
      ccgaggccg ataacagga cctttgaag tataaaatc 
      aggtcgaat ccgtcggag tttgaaact ccttcgacc 
      acggtcttg tacagttcg ccagcttcg tgtcgcgaa 
      tcgtccgat ttttggtca ggggccaat gttctgccg 
      attgagacc gggaacagc tgtccggtt aatcggtat 
      tattactcc gggtttgtg gccctgcag gcgctcttg 
      gactatacg aagataaag atcgatacg gaccgggac 
      ttccacgtg cccaaaatc agtctggag atgttccca 
      aagcacgcc cacaccggt ggttggatg gtcgctttc 
      cggctagtg atcccctta tacatggtg atggcctta 
      tctcagttc atcacctat ttactcatc ctcattgtg 
      ggcgagaaa gaaaacaag atcaaagaa gggatgaaa 
      atcatgggt ttgaaagac tccgtcttt tggctctcc 
      tggttcatt atttacggc attttcgtc ctcttctta 
      tccatcgtc tgttgcgtc ctcctcttc accctccaa 
      gtgttccaa aacaccaat ttcctcctt atcttcctt 
      ctagtcctt ttatacaca ctttcaatc atcatgttc 
      ggattcatg atcacgcca ttcttcgat aaatcgagg 
      actgccggc atcctcgga aactttgct gtgaatatc 
      atgagtttg ttttacttc atccaagtt tttatcgac 
      agttcgagt tcagtcgcg ttttgggtc gtttcgttg 
      ataagttcg tccggattc gcgttggct atggataag 
      gctttagtc atggaattg aagggtgaa ggagtcaat 
      tttgacaat ctctggtca ggacctggc atgccattc 
      ggtggtagt ttaatcatg atggctttg gacattgtc 
      ctttatgga ctcctggcc tactacctc gactgtgtg 
      atacccagc gaacatggt ataagacgt tcgcccttc 
      ttctgcttc aaacccttc ttctggttt aataagaaa 
      ccggttcaa aggatacca ctggccaat ggtgggtca 
      gctgggtcc ttaaccacc gccgaggat ggccataac 
      gacgtagag ccggtttcg agagaaatg aaaggtcgc 
      gaagccatc cgaatagtg gatatgttc aaaacgttc 
      catcattgc cggaagccc gaaattaag gcaatcaac 
      ggaatcaac ttgacgatc tacgaaggc caaatcacg 
      gccattttg gggcacaat ggtgccggc aaaaccaca 
      ctttttaac atattgaca ggtctgaca gcaccgact 
      tcgggaacg gcttacatt ttcgggtac gatgttaga 
      gaccctaac gacatggac gagatcagg agaatgaca 
      ggggtttgc ccccaacat gatatactg ttcgataac 
      ttaacccca aaagagcat ttggagttt ttcgccgcg 
      gttaaaggt attccaccg aatctgcgc gagtttgaa 
      gtgatgaaa acgttgcgt gatattgac ctgacggac 
      aaagccaac gcgtcagcc aaacattta agtggcggc 
      caaaaacgc aaactctcg attggcatt gcagtgatc 
      ggggaccca aagattata attttggac gagcccacg 
      gccggagtg gacccgtac tcgcgccgc catatgtgg 
      tcagtgctc cagaaccgt cgccatggc aaagtgatc 
      ctccttact acccatttt atggacgaa gccgatatt 
      ttggccgat aggaaagct gtggtgtcc aaaggcaac 
      atccggtgc tgtggcagc tctctcttc cttaaaaac 
      aagttcggg atcggttat cacctgacc ttggtcctt 
      gacgggatt tgtcgggaa catgccatc acccgattg 
      gtcacttca catgtcccc aaagccgaa aaagcacgt 
      cgtcatggt cgcgaactg agtttcatt ttgccacat 
      aatgctgtt gataatttc gcatcgtta ttttcggca 
      attgaacaa gaaattaac aacaagtcg agtaaactg 
      gggatttcg agctatggc gtgtcgatg acgacgctc 
      gaggaggtg ttcctgcac ttggagcgt gacgaagaa 
      actgaggga acgatggat aacttgagc aagaagatg 
      gtgaggaat cgggcgctg agtcgcagt ttgagcctc 
      cagagcaaa agtacaagt taccaatca ttacaaaac 
      gaagggaac aatgcgaca aatcaggat ggtaaaggt 
      ggcggagac acggcacag atcggcggc ttggaaatc 
      atcagtgaa accaaatcg ccaatcacc ggaattggg 
      cttgaaaaa atcgaatgc caccccaac gttttccaa 
      accttgatt tccctcctg cgtttgcgc atccttcgc 
      atgatccga gacatccag aaactctat ttcatggtg 
      ctcctgcca ctaggactc gctgctctc gttctctac 
      ttcaacagc atgcaaacc attgaagtt aaactcaca 
      ccattggcc ctcaacggc tcgacatac gccgataaa 
      acgactttt gcgatacat aacgcctca ggagtggtt 
      ttggacgac ttctacgac cagttattg gaagtaggg 
      gtccagagg attgatcca tacgatggc aatttctcg 
      ctccttctg gacgttgct ccgcacatg gctgcgtta 
      aatgtgaac gttttcaca aatgatttg agtattact 
      gtgatttac aacgatacg gcccaacac agtttgccc 
      attattttg aatttgatc agtaatggg ttgtataga 
      cttatgagt ccagagcgg gtgattaga ggacagtgg 
      gaacctatt gaagtgttg acgttaccg tttcaacaa 
      acggcacaa cctgaagag ttcgatatt gggattttt 
      tcagcgtct accttcatt gggatgatt tttgtcctg 
      gtgccagtc agtttggcc atcgatatg gtctacgat 
      cgtgagatc aaggcgaaa aaccagctg cgcgtcaac 
      ggactctcc ttctgtatg tacttcgtg acgtatttt 
      gtggtcctt ggtggcctc atgatcctg atttgtgcc 
      gcccttgtc ctaattatc cttctgttt aacattccg 
      gcgttcaaa ggctggcct gccttggtc actctcgga 
      accctaact ctcttatat tgcccttcg tccatcctt 
      ttcagtact tgtgtcagt tatatcttc gataagacg 
      gattcggct cagtcaatt ttgccaaac attgcgact 
      ttcgtcggc tgtatccct tttttcctc gtcacagct 
      ttggatatg ttgaatatt gggggaaag gcggccttt 
      gtgttacat attttattt tcgctactc aacgccatg 
      tacataccg tatgccatt atctactac atacagagg 
      gtctacata atgtgtaaa ctcaactcg acttgcaac 
      actttaacg ttgacggac tacatgacc agtgagatt 
      attgtaatg ctggtgggg atcatcgta aatattcct 
      ttctggttc ttcgtgttg atgataatt gatattaag 
      aaaagtggc ggacgtatc agtgatgcc ttcaagttt 
      ttcaaacgt aacaccgac gaaaacacc gaagacatc 
      aacgaactc agcgacatc ggcgagaac gaagaccaa 
      gacgtgaag gcggaacgt caaaaagtg aaaaacctg 
      atggactgt catgtggtg aacccaccc gttgtcatg 
      gtccaaaac ctccacaag gagtacacg aaaagcgaa 
      atcaagtgc gagtgttgt tgcaagcat gacgatgaa 
      cccgacaca accaaaatc gcaattcga agtctttcg 
      ttggctgtg gacgctggt gaagttttc ggtcttttg 
      ggacacaat ggtgctggg aaaaccacc accatgaaa 
      ataatcaca gcggaagag gccccaaca agggggcgc 
      gtacaaatc gggggcgaa aatatcacc tcgaatatg 
      aacaacgcg tttcagttg ttgggttat tgtccacaa 
      catgacgcc ttgtggaag aatatcaca gtgcgagag 
      catctagag tgctatgcg gcgattcga ggcgttccc 
      tcgggcgat attccgcaa atcgtcgat ttgtacctc 
      actgggctc caaatccac gaacatgcc gataagcag 
      gcgcagcaa tgttctggg ggcacccgc cgcaagctc 
      tccttcgcc atggcaatg gtggggaat ccgaaagtt 
      gtgcttttg gacgagccg agcactgga atggaccca 
      cggagcaaa cgctttttg tgggacacc atcttggcc 
      agtttccag ggttcacga ggcgccatc ttgaccacg 
      cactccatg gaagaagcc gatgctcta tgttcaaga 
      gttggaatt atggttaag ggggaattg aggtgtttg 
      ggatccact caacacttg aagaacctt tatggggcg 
      ggctatacg cttgaaatg aagttgagg ggtggggat 
      agaacgcca acttcgact tcttgtgac cgccccaat 
      gaactccgg gagtttgta accggattg tttcatgat 
      gcgacactg caggagagt tttgcggac agactcgtc 
      ttcagtgtt ccacaacaa agtgttcct tctttagct 
      aactgtttc atgcaactt gagaaagtt aaaacggag 
      ctggacatt gaggagtac agcttcagc caaacgacg 
      ttggagcaa gtgtttttg aaattcgcg caatacgat 
      gaagtcaac ggtgaataa
    mRNA sequence
    • >TC015859 RA
      tcagtgccg cgatcgcat cgcggccac agctcccgt 
      tctggtgcc gttttcgct tgataatat cataaaaaa 
      atcgccaaa atcgtctcg gacccgtgc aaaatgtcc 
      gcgaaaacc ggccccagt gtcagtccg ccgctgcga 
      taaaatcca atatcggtg tgtggtctt gttcggcgg 
      cgatcgacc gcccccacg ggaaaatgg cggcggtgg 
      tggcggtgt aaacgcaat gtgcaactt ttgacatag 
      ggcagtgat atttcggcg aaaaatggc atcatggtt 
      ttgattttt ccagaaatg cgtaaaaac ttgtgagcg 
      ctcgaggag tttgagaca tgggggaga cgcgaggct 
      ccatctact ttgctcagc tgaaggcca tgctgaagc 
      ggaatattc tgctgaaga agagggaaa agaggaaaa 
      cgactgcgg aggtcttgt tgcctttgt actcgcttg 
      gcgtcctca tcgtgatga aagtaatga ttccaaacc 
      caaatttcc ccgcaatag acacgccac gaggtgaag 
      tccatctct tctcctatt tccaaaaac tgaaagaac 
      acaccgtcg ccgtagttc ccaacacta acgaaacac 
      aagactttt taaaacgcg tcaacgact tgtggcaca 
      ccatcgacc acaacacgt caggttttc atccaatca 
      aatggctac agttcccaa ccgaagagg atctactta 
      ctgcatatt ggtcacaac cggacagta taccaatcg 
      ctgtgattt tcgaagaac cgaggccga taacaggac 
      ctttgaagt ataaaatca ggtcgaatc cgtcggagt 
      ttgaaactc cttcgacca cggtcttgt acagttcgc 
      cagcttcgt gtcgcgaat cgtccgatt tttggtcag 
      gggccaatg ttctgccga ttgagaccg ggaacagct 
      gtccggtta atcggtatt attactccg ggtttgtgg 
      ccctgcagg cgctcttgg actatacga agataaaga 
      tcgatacgg accgggact tccacgtgc ccaaaatca 
      gtctggaga tgttcccaa agcacgccc acaccggtg 
      gttggatgg tcgctttcc ggctagtga tccccttat 
      acatggtga tggccttat ctcagttca tcacctatt 
      tactcatcc tcattgtgg gcgagaaag aaaacaaga 
      tcaaagaag ggatgaaaa tcatgggtt tgaaagact 
      ccgtctttt ggctctcct ggttcatta tttacggca 
      ttttcgtcc tcttcttat ccatcgtct gttgcgtcc 
      tcctcttca ccctccaag tgttccaaa acaccaatt 
      tcctcctta tcttccttc tagtccttt tatacacac 
      tttcaatca tcatgttcg gattcatga tcacgccat 
      tcttcgata aatcgagga ctgccggca tcctcggaa 
      actttgctg tgaatatca tgagtttgt tttacttca 
      tccaagttt ttatcgaca gttcgagtt cagtcgcgt 
      tttgggtcg tttcgttga taagttcgt ccggattcg 
      cgttggcta tggataagg ctttagtca tggaattga 
      agggtgaag gagtcaatt ttgacaatc tctggtcag 
      gacctggca tgccattcg gtggtagtt taatcatga 
      tggctttgg acattgtcc tttatggac tcctggcct 
      actacctcg actgtgtga tacccagcg aacatggta 
      taagacgtt cgcccttct tctgcttca aacccttct 
      tctggttta ataagaaac cggttcaaa ggataccac 
      tggccaatg gtgggtcag ctgggtcct taaccaccg 
      ccgaggatg gccataacg acgtagagc cggtttcga 
      gagaaatga aaggtcgcg aagccatcc gaatagtgg 
      atatgttca aaacgttcc atcattgcc ggaagcccg 
      aaattaagg caatcaacg gaatcaact tgacgatct 
      acgaaggcc aaatcacgg ccattttgg ggcacaatg 
      gtgccggca aaaccacac tttttaaca tattgacag 
      gtctgacag caccgactt cgggaacgg cttacattt 
      tcgggtacg atgttagag accctaacg acatggacg 
      agatcagga gaatgacag gggtttgcc cccaacatg 
      atatactgt tcgataact taaccccaa aagagcatt 
      tggagtttt tcgccgcgg ttaaaggta ttccaccga 
      atctgcgcg agtttgaag tgatgaaaa cgttgcgtg 
      atattgacc tgacggaca aagccaacg cgtcagcca 
      aacatttaa gtggcggcc aaaaacgca aactctcga 
      ttggcattg cagtgatcg gggacccaa agattataa 
      ttttggacg agcccacgg ccggagtgg acccgtact 
      cgcgccgcc atatgtggt cagtgctcc agaaccgtc 
      gccatggca aagtgatcc tccttacta cccatttta 
      tggacgaag ccgatattt tggccgata ggaaagctg 
      tggtgtcca aaggcaaca tccggtgct gtggcagct 
      ctctcttcc ttaaaaaca agttcggga tcggttatc 
      acctgacct tggtccttg acgggattt gtcgggaac 
      atgccatca cccgattgg tcacttcac atgtcccca 
      aagccgaaa aagcacgtc gtcatggtc gcgaactga 
      gtttcattt tgccacata atgctgttg ataatttcg 
      catcgttat tttcggcaa ttgaacaag aaattaaca 
      acaagtcga gtaaactgg ggatttcga gctatggcg 
      tgtcgatga cgacgctcg aggaggtgt tcctgcact 
      tggagcgtg acgaagaaa ctgagggaa cgatggata 
      acttgagca agaagatgg tgaggaatc gggcgctga 
      gtcgcagtt tgagcctcc agagcaaaa gtacaagtt 
      accaatcat tacaaaacg aagggaaca atgcgacaa 
      atcaggatg gtaaaggtg gcggagaca cggcacaga 
      tcggcggct tggaaatca tcagtgaaa ccaaatcgc 
      caatcaccg gaattgggc ttgaaaaaa tcgaatgcc 
      accccaacg ttttccaaa ccttgattt ccctcctgc 
      gtttgcgca tccttcgca tgatccgag acatccaga 
      aactctatt tcatggtgc tcctgccac taggactcg 
      ctgctctcg ttctctact tcaacagca tgcaaacca 
      ttgaagtta aactcacac cattggccc tcaacggct 
      cgacatacg ccgataaaa cgacttttg cgatacata 
      acgcctcag gagtggttt tggacgact tctacgacc 
      agttattgg aagtagggg tccagagga ttgatccat 
      acgatggca atttctcgc tccttctgg acgttgctc 
      cgcacatgg ctgcgttaa atgtgaacg ttttcacaa 
      atgatttga gtattactg tgatttaca acgatacgg 
      cccaacaca gtttgccca ttattttga atttgatca 
      gtaatgggt tgtatagac ttatgagtc cagagcggg 
      tgattagag gacagtggg aacctattg aagtgttga 
      cgttaccgt ttcaacaaa cggcacaac ctgaagagt 
      tcgatattg ggatttttt cagcgtcta ccttcattg 
      ggatgattt ttgtcctgg tgccagtca gtttggcca 
      tcgatatgg tctacgatc gtgagatca aggcgaaaa 
      accagctgc gcgtcaacg gactctcct tctgtatgt 
      acttcgtga cgtattttg tggtccttg gtggcctca 
      tgatcctga tttgtgccg cccttgtcc taattatcc 
      ttctgttta acattccgg cgttcaaag gctggcctg 
      ccttggtca ctctcggaa ccctaactc tcttatatt 
      gcccttcgt ccatccttt tcagtactt gtgtcagtt 
      atatcttcg ataagacgg attcggctc agtcaattt 
      tgccaaaca ttgcgactt tcgtcggct gtatccctt 
      ttttcctcg tcacagctt tggatatgt tgaatattg 
      ggggaaagg cggcctttg tgttacata ttttatttt 
      cgctactca acgccatgt acataccgt atgccatta 
      tctactaca tacagaggg tctacataa tgtgtaaac 
      tcaactcga cttgcaaca ctttaacgt tgacggact 
      acatgacca gtgagatta ttgtaatgc tggtgggga 
      tcatcgtaa atattcctt tctggttct tcgtgttga 
      tgataattg atattaaga aaagtggcg gacgtatca 
      gtgatgcct tcaagtttt tcaaacgta acaccgacg 
      aaaacaccg aagacatca acgaactca gcgacatcg 
      gcgagaacg aagaccaag acgtgaagg cggaacgtc 
      aaaaagtga aaaacctga tggactgtc atgtggtga 
      acccacccg ttgtcatgg tccaaaacc tccacaagg 
      agtacacga aaagcgaaa tcaagtgcg agtgttgtt 
      gcaagcatg acgatgaac ccgacacaa ccaaaatcg 
      caattcgaa gtctttcgt tggctgtgg acgctggtg 
      aagttttcg gtcttttgg gacacaatg gtgctggga 
      aaaccacca ccatgaaaa taatcacag cggaagagg 
      ccccaacaa gggggcgcg tacaaatcg ggggcgaaa 
      atatcacct cgaatatga acaacgcgt ttcagttgt 
      tgggttatt gtccacaac atgacgcct tgtggaaga 
      atatcacag tgcgagagc atctagagt gctatgcgg 
      cgattcgag gcgttccct cgggcgata ttccgcaaa 
      tcgtcgatt tgtacctca ctgggctcc aaatccacg 
      aacatgccg ataagcagg cgcagcaat gttctgggg 
      gcacccgcc gcaagctct ccttcgcca tggcaatgg 
      tggggaatc cgaaagttg tgcttttgg acgagccga 
      gcactggaa tggacccac ggagcaaac gctttttgt 
      gggacacca tcttggcca gtttccagg gttcacgag 
      gcgccatct tgaccacgc actccatgg aagaagccg 
      atgctctat gttcaagag ttggaatta tggttaagg 
      gggaattga ggtgtttgg gatccactc aacacttga 
      agaaccttt atggggcgg gctatacgc ttgaaatga 
      agttgaggg gtggggata gaacgccaa cttcgactt 
      cttgtgacc gccccaatg aactccggg agtttgtaa 
      ccggattgt ttcatgatg cgacactgc aggagagtt 
      ttgcggaca gactcgtct tcagtgttc cacaacaaa 
      gtgttcctt ctttagcta actgtttca tgcaacttg 
      agaaagtta aaacggagc tggacattg aggagtaca 
      gcttcagcc aaacgacgt tggagcaag tgtttttga 
      aattcgcgc aatacgatg aagtcaacg gtgaataaa 
      gccgtattg tatcatgtg atttagata agttgaaat 
      atctcattt gaaagaatc aaaattgat tatttttta 
      actcactaa aatgttgag tagtttaaa tgtgttgac 
      ttaagttga aattgtaca gttttaaac ttataacgt 
      aattaaaca gaatgtata gtttgtgta tgcatgact 
      gttgtatga aaattgaat ttaattcgg ttgtcttgt 
      tggcctaaa ctttttttc cattttttt taacttgat 
      gataatagt gcctatcgt atgttatgt aaatagtcg 
      cttgcacag acttgtcgc ttgcagttt gtttcatta 
      gtgaaaatt tttaaataa agtgttcag ttattatgg 
      caacggtta cgatctgtt tgcccattt ggacgacaa 
      tttttactt tctccgcaa taatgcaaa tgtatcaat 
      taaattgga ataaaattt tattgcgaa a
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