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Gene information
General information
TC003483 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: 5844476..5858713
Sequence information
Transcripts / Proteins (2) 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
    TC003483
    iBeetle screen iB_08892

    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: < 20
    Phenotype after larval injection
    Usually 10 injected animals

    Larval injection procedure

    Day 0: 10 female L5-6 larvae of the D17Xhom strain (females express RFP in the eyes and in the CNS; pupae express GFP in the thorax) were injected with dsRNA.

    11 days post injection (dpi): Pupal morphology Documentation of larval/ pupal death and defects of metamorphosis (eclosion, delayed development). Screening for alterations of pupal head and appendages, pupal thorax, pupal thoracic musculature, pupal legs, pupal wings, pupal abdomen and pupal genital lobes.

    16 dpi: Adult morphology Documentation of larval/ pupal and adult death and defects of metamorphosis (eclosion, delayed development). Screening for alterations of adult head and appendages, adult thorax, adult elytra and hindwings, adult legs and cuticle properties. For mating 4 males of the black strain were added.

    19 dpi: Sieving and egg-lay Documentation of adult lethality and enduring metamorphosis defects.

    22 dpi: Ovary analysis (Fertility) In case of a reduction of egg production 4 females were dissected and the gross morphology of the ovaries were analysed.

    41 dpi: Fertility and stink gland analysis Documentation of defects in abdominal and thoracic stink glands development (colour, size, content) of the injected females. Check for fertility of injected larvae (no or decreased offspring).

    Terms used in the larval injection procedure

    pupal/ adult pantagmatic defects
    At least two tagmata (head, thorax, abdomen or terminus) show similar/ comparable defects.
    pupal molt not fulfilled
    The hatching from the larva to the pupa is interrupted. Either it dies as prepupa or the phenotype shows larval as well as pupal features.
    eclosion not fulfilled
    The emergence of the adult from the pupa stage is interrupted. This phenotype shows pupal as well as adult features.
    starved
    Dissected females show a strong resorption of fatbody predominately in the abdomen and the oogenesis is blocked. These are considered as eventually lethal phenotypes and the blocked oogenesis probably a secondary defect due to starvation.
    Metamorphosis and survival

    Lethalities 11 days after larval injection: % (includes death as larva, prepupa, pupa)
    Lethalities 22 days after larval injection: % (includes death as larva, prepupa, pupa, adult)

    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC003483 RA
      MKKQNVRTL SLIVCTFTY LLIGAAVFD ALESKTENQ 
      RKVVLQEIQ AIVRRKYNI SDEDFRIME TVVLKSEPH 
      RAGQQWKFT GAFYYATTV LTTIGYGHS TPSTIAGKL 
      FTMCYAMIG IPLGLVMFQ SIGERVNKL SSVIIRSVK 
      SSLHCRQTA ASELDLICV VTTLSSLTI AGGAAAFSR 
      YEGWSYFDS VYYCFITLT TIGFGDMVA LQKDNALSK 
      KPEYVMFAL IFILFGLAI VAASLNLLV LRFVTMNTE 
      DERRDEAQA MQALAGAVR LEGDVITAN GSILSGQMG 
      HHCTETTSL DADDTSVCS CHCGCFTSN SRHRSLSRF 
      TVRRSPTQI GHLLPVLPM HELHLPQGK ILPPPAIYP 
      NHRASI
    • >TC003483 RB
      MKKQNVRTL SLIVCTFTY LLIGAAVFD ALESKTENQ 
      RKVVLQEIQ AIVRRKYNI SDEDFRIME TVVLKSEPH 
      RAGQQWKFT GAFYYATTV LTTIGYGHS TPSTIAGKL 
      FTMCYAMIG IPLGLVMFQ SIGERVNKL SSVIIRSVK 
      SSLHCRQTA ASELDLICV VTTLSSLTI AGGAAAFSR 
      YEGWSYFDS VYYCFITLT TIGFGDMVA LQKDNALSK 
      KPEYVMFAL IFILFGLAI VAASLNLLV LRFVTMNTE 
      DERRDEAQA MQALAGAVR LEGDVITAN GSILSGQMG 
      HHCTETTSL DADDTSVCS CHCGCFTSN SRHRFTVRR 
      SPTQIGHLL PVLPMHELH LPQGKILPP PAIYPNHRA 
      SI
    CDS sequence
    • >TC003483 RA
      atgaagaag caaaacgtt cgcacgcta tctcttatc 
      gtttgcacc ttcacgtac ctgctgatc ggcgccgcc 
      gtcttcgac gctcttgaa tctaaaacc gaaaatcaa 
      cgcaaagtc gtcctgcaa gaaatccaa gcaatagtt 
      agaagaaaa tacaacata agcgacgaa gatttcagg 
      ataatggaa actgtcgtc ctgaaatct gagccgcat 
      agagccggc caacaatgg aaattcact ggagccttc 
      tactacgcg actactgtt ctaaccacg ataggttac 
      ggccactct acacccagc accattgcg gggaagctg 
      ttcaccatg tgttacgct atgataggg ataccgcta 
      ggactcgtc atgttccag agcataggc gagagggtc 
      aacaagcta agcagtgtg ataattcgg tccgtgaag 
      tcttccttg cactgccga caaacagca gcctccgag 
      ctggacctc atctgtgtc gtaacaacg ctgagttcg 
      ttgacgata gctggtggt gctgccgcc ttctcgcgt 
      tacgaaggc tggagctac ttcgactcc gtttattat 
      tgcttcatc acactcact actatagga ttcggtgac 
      atggtagca ctccaaaaa gacaatgct ctcagtaaa 
      aaaccggag tatgtcatg tttgcactc atcttcatc 
      ctctttggc ttggccatt gtggcggct tcactgaac 
      ttgctcgtt cttcgcttc gtcacgatg aacacggaa 
      gacgagagg cgggacgag gcgcaagcc atgcaggct 
      ctggctggc gccgtgaga ctcgaaggc gacgtcatc 
      acggccaat ggctcgatt ctgagcggc caaatgggc 
      catcactgt acagaaacc acctcccta gacgctgat 
      gatacctcc gtgtgttct tgtcattgt ggctgtttt 
      acttctaat agtaggcac aggtcgctt tccaggttc 
      acagtacga aggtctccc acgcaaatc gggcatttg 
      ctccccgtt ctgccaatg catgagctt catcttccc 
      caagggaaa atattgccc ccaccggcg atatacccc 
      aaccatagg gcttcgata tga
    • >TC003483 RB
      atgaagaag caaaacgtt cgcacgcta tctcttatc 
      gtttgcacc ttcacgtac ctgctgatc ggcgccgcc 
      gtcttcgac gctcttgaa tctaaaacc gaaaatcaa 
      cgcaaagtc gtcctgcaa gaaatccaa gcaatagtt 
      agaagaaaa tacaacata agcgacgaa gatttcagg 
      ataatggaa actgtcgtc ctgaaatct gagccgcat 
      agagccggc caacaatgg aaattcact ggagccttc 
      tactacgcg actactgtt ctaaccacg ataggttac 
      ggccactct acacccagc accattgcg gggaagctg 
      ttcaccatg tgttacgct atgataggg ataccgcta 
      ggactcgtc atgttccag agcataggc gagagggtc 
      aacaagcta agcagtgtg ataattcgg tccgtgaag 
      tcttccttg cactgccga caaacagca gcctccgag 
      ctggacctc atctgtgtc gtaacaacg ctgagttcg 
      ttgacgata gctggtggt gctgccgcc ttctcgcgt 
      tacgaaggc tggagctac ttcgactcc gtttattat 
      tgcttcatc acactcact actatagga ttcggtgac 
      atggtagca ctccaaaaa gacaatgct ctcagtaaa 
      aaaccggag tatgtcatg tttgcactc atcttcatc 
      ctctttggc ttggccatt gtggcggct tcactgaac 
      ttgctcgtt cttcgcttc gtcacgatg aacacggaa 
      gacgagagg cgggacgag gcgcaagcc atgcaggct 
      ctggctggc gccgtgaga ctcgaaggc gacgtcatc 
      acggccaat ggctcgatt ctgagcggc caaatgggc 
      catcactgt acagaaacc acctcccta gacgctgat 
      gatacctcc gtgtgttct tgtcattgt ggctgtttt 
      acttctaat agtaggcac aggttcaca gtacgaagg 
      tctcccacg caaatcggg catttgctc cccgttctg 
      ccaatgcat gagcttcat cttccccaa gggaaaata 
      ttgccccca ccggcgata taccccaac catagggct 
      tcgatatga
    mRNA sequence
    • >TC003483 RA
      gctggcgtc tccacatcc agaagccga gtggattcg 
      gagcaccgt cgtgacgtc actgcgacg tcgttatcg 
      atctcttaa ccgcgtgtt acaacagaa agccccgaa 
      aagctctag tacacggtc ggcggtggt gtttgtgcc 
      aaaatttgc aaagtcgtc ttctgcact gcagcgcac 
      tcgacaatg gttgctgtc cgctcggaa aagagcgta 
      tgtgtctcg tcataaaaa tcgctttgg acgttcctc 
      ctcggtttg cagattacc tcgctattt tagtgcggg 
      gttgtgatg tggttgact ggagggggg tgcgcgggg 
      ggcagctag cggcgtggg agccccccg gccccagca 
      tgacgcgcg ctagagaat gaagaagca aaacgttcg 
      cacgctatc tcttatcgt ttgcacctt cacgtacct 
      gctgatcgg cgccgccgt cttcgacgc tcttgaatc 
      taaaaccga aaatcaacg caaagtcgt cctgcaaga 
      aatccaagc aatagttag aagaaaata caacataag 
      cgacgaaga tttcaggat aatggaaac tgtcgtcct 
      gaaatctga gccgcatag agccggcca acaatggaa 
      attcactgg agccttcta ctacgcgac tactgttct 
      aaccacgat aggttacgg ccactctac acccagcac 
      cattgcggg gaagctgtt caccatgtg ttacgctat 
      gatagggat accgctagg actcgtcat gttccagag 
      cataggcga gagggtcaa caagctaag cagtgtgat 
      aattcggtc cgtgaagtc ttccttgca ctgccgaca 
      aacagcagc ctccgagct ggacctcat ctgtgtcgt 
      aacaacgct gagttcgtt gacgatagc tggtggtgc 
      tgccgcctt ctcgcgtta cgaaggctg gagctactt 
      cgactccgt ttattattg cttcatcac actcactac 
      tataggatt cggtgacat ggtagcact ccaaaaaga 
      caatgctct cagtaaaaa accggagta tgtcatgtt 
      tgcactcat cttcatcct ctttggctt ggccattgt 
      ggcggcttc actgaactt gctcgttct tcgcttcgt 
      cacgatgaa cacggaaga cgagaggcg ggacgaggc 
      gcaagccat gcaggctct ggctggcgc cgtgagact 
      cgaaggcga cgtcatcac ggccaatgg ctcgattct 
      gagcggcca aatgggcca tcactgtac agaaaccac 
      ctccctaga cgctgatga tacctccgt gtgttcttg 
      tcattgtgg ctgttttac ttctaatag taggcacag 
      gtcgctttc caggttcac agtacgaag gtctcccac 
      gcaaatcgg gcatttgct ccccgttct gccaatgca 
      tgagcttca tcttcccca agggaaaat attgccccc 
      accggcgat ataccccaa ccatagggc ttcgatatg 
      atttcctgg ggcataagc ctacatgtt caatgtgat 
      acgtaaaaa ctagataaa agtaatcaa ttcgaacga 
      aacgttagt ggtttttaa tcatcgatt gtgtgtaaa 
      gatgagagt cgtctcatt ttatgaaac aataaccat 
      ataatcgca ctaactgaa acgtcttat gaaacaagt 
      atgaaagca caaatcgat attttttct taaggatta 
      tttctatga tttacgtgt ttctcacaa tcagtgaat 
      ttgttggaa gcgcagttt gtatacaaa ttgatgtga 
      tttcgatgt aaatgccag ctcctagtt tcatccccg 
      agaaggtat ccgttgtct tttccatta ttgtagcgt 
      gatactttg gacacacac aaagttatt atacgggtt 
      gagagtcac ccacgaaat ttgggatca aagtaacgt 
      agttgccaa atttatctc gtttcttat acacttttg 
      aaaacttcc aatatctaa taatatcga gttactaag 
      ccctcaaat cttccatga tttatcaaa tgttaaatg 
      tccacgttt catatctga cggcgaatg caacttttg 
      actgttgtt aagcggaaa ttaaattac agttcgtta 
      tcatttgct tcgtcgaga attcttaca tctgcttct 
      atcaaactt tcgttgact cgaagggaa cataagtag 
      cgttactgt tactaaact aaaacgttt ctaaatagt 
      tttcaaaag tgtgaacaa taagtaaat gttcttccg 
      tttccttga aacagttgc ttctattac ttgccaaaa 
      accgacact agataagtg gtgcttcct gaacaatgc 
      tatttaccg cactgagct taagggcct ctgaaacac 
      ccttaataa atgttaaaa atctgctca agtaccatt 
      gcagtattg tttggttcg aatactcag tattagcat 
      cgcaagatg gactaacgt aaggacaca ctgggcaga 
      ttttcaaag tgcattgta aataaccag actgcattg 
      tacataatc cgagtactg atttagtca tttttgaaa 
      acttagtct tgatttgag attgtacgt agacttttg 
      ataaaaatt gttttaaag aaagaacat gtctaatcg 
      ataaatttc catgatggt tatgaattt taggcctaa 
      cattaacat aacagtatt gcgtagatt ttacacttt 
      taccacaaa acgttgaat atgatatag acatagagt 
      acatttcta aattttaat cgaggtcat tttaataaa 
      gagttgttc tttctgtta aaacttatg taagtaatg 
      tataatttt tgaatcata gtatgtaat tatacttaa 
      aaaatgtgt caaccattt gaaatgtta cctattaat 
      taagcttgt ttttattga aaaattgta actgctgaa 
      cggtttaga taggttcaa gttgctgta aataccaca 
      taaaaatga caaagaata ggttgtaaa ggtgtacgt 
      atacatagt ctttccgcc aaatcactg tcgataatt 
      atttttcga attatatca ccgaacact ttactaaat 
      acccatagg ctaacattt ccacacttc tactgatga 
      aatagattt atgtaaaaa ttgtcatct agtttttta 
      atgtcagtg tttgcatgt tgaacagat agatttacc 
      tcacagagc aagataaaa ctgcgcttt tcttcattt 
      cttgtctta ttatttttc gactgtcat tgtgatgct 
      aaaaacgag cgttaaaaa ggaccatgt gtaaataaa 
      cttttacta aacca
    • >TC003483 RB
      gctggcgtc tccacatcc agaagccga gtggattcg 
      gagcaccgt cgtgacgtc actgcgacg tcgttatcg 
      atctcttaa ccgcgtgtt acaacagaa agccccgaa 
      aagctctag tacacggtc ggcggtggt gtttgtgcc 
      aaaatttgc aaagtcgtc ttctgcact gcagcgcac 
      tcgacaatg gttgctgtc cgctcggaa aagagcgta 
      tgtgtctcg tcataaaaa tcgctttgg acgttcctc 
      ctcggtttg cagattacc tcgctattt tagtgcggg 
      gttgtgatg tggttgact ggagggggg tgcgcgggg 
      ggcagctag cggcgtggg agccccccg gccccagca 
      tgacgcgcg ctagagaat gaagaagca aaacgttcg 
      cacgctatc tcttatcgt ttgcacctt cacgtacct 
      gctgatcgg cgccgccgt cttcgacgc tcttgaatc 
      taaaaccga aaatcaacg caaagtcgt cctgcaaga 
      aatccaagc aatagttag aagaaaata caacataag 
      cgacgaaga tttcaggat aatggaaac tgtcgtcct 
      gaaatctga gccgcatag agccggcca acaatggaa 
      attcactgg agccttcta ctacgcgac tactgttct 
      aaccacgat aggttacgg ccactctac acccagcac 
      cattgcggg gaagctgtt caccatgtg ttacgctat 
      gatagggat accgctagg actcgtcat gttccagag 
      cataggcga gagggtcaa caagctaag cagtgtgat 
      aattcggtc cgtgaagtc ttccttgca ctgccgaca 
      aacagcagc ctccgagct ggacctcat ctgtgtcgt 
      aacaacgct gagttcgtt gacgatagc tggtggtgc 
      tgccgcctt ctcgcgtta cgaaggctg gagctactt 
      cgactccgt ttattattg cttcatcac actcactac 
      tataggatt cggtgacat ggtagcact ccaaaaaga 
      caatgctct cagtaaaaa accggagta tgtcatgtt 
      tgcactcat cttcatcct ctttggctt ggccattgt 
      ggcggcttc actgaactt gctcgttct tcgcttcgt 
      cacgatgaa cacggaaga cgagaggcg ggacgaggc 
      gcaagccat gcaggctct ggctggcgc cgtgagact 
      cgaaggcga cgtcatcac ggccaatgg ctcgattct 
      gagcggcca aatgggcca tcactgtac agaaaccac 
      ctccctaga cgctgatga tacctccgt gtgttcttg 
      tcattgtgg ctgttttac ttctaatag taggcacag 
      gttcacagt acgaaggtc tcccacgca aatcgggca 
      tttgctccc cgttctgcc aatgcatga gcttcatct 
      tccccaagg gaaaatatt gcccccacc ggcgatata 
      ccccaacca tagggcttc gatatgatt tcctggggc 
      ataagccta catgttcaa tgtgatacg taaaaacta 
      gataaaagt aatcaattc gaacgaaac gttagtggt 
      ttttaatca tcgattgtg tgtaaagat gagagtcgt 
      ctcatttta tgaaacaat aaccatata atcgcacta 
      actgaaacg tcttatgaa acaagtatg aaagcacaa 
      atcgatatt ttttcttaa ggattattt ctatgattt 
      acgtgtttc tcacaatca gtgaatttg ttggaagcg 
      cagtttgta tacaaattg atgtgattt cgatgtaaa 
      tgccagctc ctagtttca tccccgaga aggtatccg 
      ttgtctttt ccattattg tagcgtgat actttggac 
      acacacaaa gttattata cgggttgag agtcaccca 
      cgaaatttg ggatcaaag taacgtagt tgccaaatt 
      tatctcgtt tcttataca cttttgaaa acttccaat 
      atctaataa tatcgagtt actaagccc tcaaatctt 
      ccatgattt atcaaatgt taaatgtcc acgtttcat 
      atctgacgg cgaatgcaa cttttgact gttgttaag 
      cggaaatta aattacagt tcgttatca tttgcttcg 
      tcgagaatt cttacatct gcttctatc aaactttcg 
      ttgactcga agggaacat aagtagcgt tactgttac 
      taaactaaa acgtttcta aatagtttt caaaagtgt 
      gaacaataa gtaaatgtt cttccgttt ccttgaaac 
      agttgcttc tattacttg ccaaaaacc gacactaga 
      taagtggtg cttcctgaa caatgctat ttaccgcac 
      tgagcttaa gggcctctg aaacaccct taataaatg 
      ttaaaaatc tgctcaagt accattgca gtattgttt 
      ggttcgaat actcagtat tagcatcgc aagatggac 
      taacgtaag gacacactg ggcagattt tcaaagtgc 
      attgtaaat aaccagact gcattgtac ataatccga 
      gtactgatt tagtcattt ttgaaaact tagtcttga 
      tttgagatt gtacgtaga cttttgata aaaattgtt 
      ttaaagaaa gaacatgtc taatcgata aatttccat 
      gatggttat gaattttag gcctaacat taacataac 
      agtattgcg tagatttta cacttttac cacaaaacg 
      ttgaatatg atatagaca tagagtaca tttctaaat 
      tttaatcga ggtcatttt aataaagag ttgttcttt 
      ctgttaaaa cttatgtaa gtaatgtat aatttttga 
      atcatagta tgtaattat acttaaaaa atgtgtcaa 
      ccatttgaa atgttacct attaattaa gcttgtttt 
      tattgaaaa attgtaact gctgaacgg tttagatag 
      gttcaagtt gctgtaaat accacataa aaatgacaa 
      agaataggt tgtaaaggt gtacgtata catagtctt 
      tccgccaaa tcactgtcg ataattatt tttcgaatt 
      atatcaccg aacacttta ctaaatacc cataggcta 
      acatttcca cacttctac tgatgaaat agatttatg 
      taaaaattg tcatctagt tttttaatg tcagtgttt 
      gcatgttga acagataga tttacctca cagagcaag 
      ataaaactg cgcttttct tcatttctt gtcttatta 
      tttttcgac tgtcattgt gatgctaaa aacgagcgt 
      taaaaagga ccatgtgta aataaactt ttactaaac 
      ca
    The data is licensed under a Creative Commons Attribution 3.0 Unported License. (CC-BY) CC-BY