| General information | |||
| TC009935 | 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_007422.5: 11593137..11604510 | |
| 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. | |||
| Closest fly homologs |
|
TC009935 | |
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.
| General information | ||||
| iB_08764 | iB fragment
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. |
Get FastA Get primers |
||
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.
Lethality 11 days after pupal injection: 30.0% (includes death as pupa, adult)
eclosion not fulfilled (with pupal and adult features) - Number of animals showing the phenotype: 2 (3 dpi)adult lethality - Number of animals showing the phenotype: 1 (9 dpi)
adult nutritional state bad (cachectic) - Number of animals showing the phenotype: 1 (11 dpi)
vitellogenic egg chamber late degenerated - Number of animals showing the phenotype: 3
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.
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)
- >TC009935 RA
MTTLELYTK GAKVWIPHP EKVWEGAEL EEDYKTNKK SLEVVTENN EKKTLTIKS ESDLPCLRN PSILVGEND LTSLSYLHE PAVLYNLQV RFCQNRYIY TYCGIVLVA INPYDDLPI YDVDTIQTY RGQAMGDLD PHIFAVAEE AYTKLEREQ RDQSIIVSG ESGAGKTVS AKYAMRYFA TVGGSATET QVEKKVLAS SPIMEAIGN AKTTRNDNS SRFGKFIEL QFNKQFHIS GASMRTYLL EKSRVVFQA PDERNYHIF YQLCSARDQ FPHLHLAHQ DSFYYLNQG ESPNVDGVD DLQTFEETL YALNLLGFK KSDLDDMFK IIASVLHLG NIKFVESII STENEQDQE GCSILGTDS HLKILAELL EIDSKEMQQ WLCTRKIVS MREVFLKPM SVEDAQAAR DALAKHIYA ELFNWIVLV INKALESDI PRHKFIGVL DIYGFETFE TNSFEQFCI NYANEKLQQ QFNLHVFKL EQEEYIKEG IEWKMIDFY DNQPCIDLI ETKLGILDL LDEECRMPR GTDSSWTEK LYSKCAKYS HFGKARFGT SAFTINHFA DKVQYESNG FLEKNRDTV IEEQISVVK RSKNDLVRR LFGDDSQKL AVPGAKLKV ISAKPAPTS QKTHKKSVG SQFRDSLNM LMTTLNATT PHYVRCIKP NDSKKPFEY NPKRAVQQL RACGVLETI RISAAGFPS RWTYVDFFY RYRVLCKFK DIKRNDMQL TCQNILNQY IKNKDMFQF GKTKIFFRA GQVAYLEKL RADKLKACC IMMQKTIRA FIWRKKYLR IKRCTINIQ RYGRGYLAR KLAENLRRE KAAKTLQRY VRGWVKRVQ YQRLKACVT GIQRYARGY LARRRYMQL RYNAKALVI QRYVRGYLA RRSALAYKR KIVICQAAI RRFLARRLY KKLRIEARS IEHVKKLNK GLENKIISL QQKIDEITK QNAELMTYK AGVNELKNK LTAFKANEA EIKNLNARL IEKDKVIDK LKEDLKLER DEKIDLINE HEKYRQETQ KQRDLWTQE TNKLRKELD NINEIVKMN QKGAEENLK VRLEEEKML ILNEQDSDR EAYQKLLQE YHCLEQHCE ELEKQLNNQ SQNQSSHRR NVSDLSSID SFVINSDIA EDHGYGSVR STTTSSSQR EKLENIDWK CEGNVDSQT PSTSSSDTK HESENNKVD VGLVLKLQH KLAEVEREK LRMQKRLDE LDMSPRTEK AENAARDSI KISELELVN SNLKAQLME LQNSIIEGT GSSKIHEQM QILHSELER KTEEIIQLK SVLADQTNN MKSLVNSNA RLGAYINED GELALAYET QKTINKQLE LELQDEKSK YKAHEREYK LEIEKLRED NERQQRLLS ANLTNTPQT QSEAFMQHE ITRLTIENL DLHDKNDTL TETVRKLKK QIKLMAKKL KEAGLDMDD NFNEITDNE PNKTNVNKH TRALPSIRK KERDYLGMF SYSSGEENT IMRQLVIDL KPRTAVTLL PGLPAYIVF MCIRHTDYV NDEDKVKAL LSAFTNSVK KVIKKRHED FETMALWLS NTLRLVHNM KQYSGDRAF QAKNTPKQN EQCLRNFDL SEYRQVLSD IAVWIYQGL IRKFAEKIQ PLVIPAILE HEEIPGISG NKPSGFRGR SSSVATSPE PSQKPTTAV LLELTNHHK ILSFYGVDP EVISQIFKQ IFYFLCATS LNNLLLRQE LCHWSKGFQ IRHNLSHFE MWTREKGLD EASIQSTLQ PIIQAAHLL QARKTEEDV ASVCEMCSA LTPLQICKI LNLYKPVDE FEQHVPPSF ISKVKAKLQ ERNPTSEQQ TLLMDVKYH FPVRFPFNP SVICLEDIE IPEVLKLPQ LEKL
- >TC009935 RA
atgactacc ttggagcta tatacaaag ggtgccaag gtatggata cctcaccct gaaaaagta tgggaaggc gctgaactt gaagaagac tacaaaacg aacaaaaag tcacttgaa gtagtaaca gaaaacaac gaaaagaaa acccttacg atcaaatca gaatcggat ttaccctgt ctgaggaac ccttcaatt ttagtcggg gaaaatgac cttacatcc ctctcctac ttacacgag cccgccgta ctttacaat ttacaagta cggttctgc caaaatcga tacatttac acctattgt gggattgtt ttggtggca attaatccc tacgatgat cttcctatt tatgacgtc gacaccata caaacgtat agaggtcaa gcgatgggt gacctagat cctcacata ttcgccgta gccgaagaa gcgtacacg aaactagaa agagagcaa agggaccaa agcataatc gtatcagga gaaagcgga gccggtaaa acggtttcc gcgaaatac gcaatgaga tacttcgcg actgtagga ggaagtgcg accgaaacc caagtcgaa aaaaaagtc ctagctagt agtcctatc atggaagcc atcggaaac gcgaaaact acaagaaac gataacagt tcgcgattt ggaaagttt atagaactc cagtttaat aagcaattt catattagc ggagcgtcg atgagaacg tacctcttg gaaaaatcg cgagtcgtc tttcaggct cccgatgaa cggaattac catattttc taccaattg tgttccgcg agagaccaa tttccacac ttgcatctc gctcaccaa gatagtttt tattattta aatcaagga gaaagtcca aatgtggac ggagtcgac gatttacaa accttcgaa gagacgctc tatgcgttg aatctttta gggtttaag aaatcggat ctggacgat atgtttaaa attatagcc tccgtcttg catttggga aatatcaag tttgttgaa tcgatcatt tccacggaa aacgaacaa gaccaggag ggttgtagc atactgggc actgactca catttgaaa atcttagcg gaattactc gagatcgat tcaaaagaa atgcagcag tggctctgt acccgaaaa atcgtctct atgagagag gtcttcctg aaaccaatg tccgtcgaa gacgcgcaa gctgcccgt gatgcttta gcaaaacac atttatgca gaattgttc aactggatc gtgttagtt ataaacaaa gctttagaa tccgacatt cctagacat aaatttatc ggagtcctc gatatctac ggtttcgaa accttcgaa accaattcc tttgaacaa ttttgtatc aattacgcc aacgaaaaa ctccaacaa caattcaac ctccacgtg tttaaactg gagcaggag gaatacatt aaagaagga attgaatgg aaaatgatt gatttctac gacaaccaa ccctgcatc gacctcatc gaaacaaaa ttagggatt ttagatttg ctagacgag gagtgtcgg atgccgcga ggtactgac agttcctgg acggagaaa ctgtactca aagtgcgca aagtattcc cacttcggc aaggcgcgt ttcggcact tctgccttc accatcaat cactttgcc gataaagtc caatacgaa agcaacggt tttctggag aaaaaccgc gacaccgta atcgaggag caaatctcg gtggtgaaa aggagcaaa aacgatcta gtccgccgg cttttcggc gacgattct caaaaattg gccgttccg ggagccaaa ctcaaagtg atctcggcc aaacctgcc ccaacctcg caaaaaacc cacaagaaa tcggtgggg tcccagttt agagactcg cttaatatg ctaatgacc acgttgaac gccacaacc ccgcattac gtccggtgt attaagcca aacgattcg aaaaaaccg tttgagtat aacccgaaa cgggcggtg caacaactg cgagcttgt ggcgtgttg gagacgatc aggatttcg gcagccggg tttccgagt cggtggact tacgttgat ttcttctac cggtaccga gtcttgtgc aagtttaaa gatatcaaa agaaacgac atgcagctc acttgccag aatatactg aaccagtat attaaaaac aaggatatg tttcaattc gggaaaact aagattttt ttccgggcc ggacaagtc gcctatttg gagaaatta agagccgat aaactcaaa gcctgttgc atcatgatg caaaaaaca atccgggcg ttcatttgg aggaaaaag tacctccga atcaagcgc tgcactatc aatatacaa cggtatgga cggggttat ttggcccga aaactggcc gaaaatttg cggcgcgag aaagccgca aagactttg cagagatat gttcgagga tgggtgaaa cgtgtccaa tatcaaaga ctgaaggcc tgtgtgact ggcattcaa aggtatgcc agagggtat ttagcgaga cggcggtac atgcaatta cggtacaac gccaaagct ctggtaata cagcgatat gtaagaggg tatttagca cggagaagt gccctggct tacaagcgg aaaatcgtc atatgccaa gctgcgatt cgccgcttt ttggctaga agattatac aagaaactc agaatcgaa gcaagatca atcgaacac gtcaagaaa cttaacaaa ggcttggag aacaagatc atcagtttg cagcaaaag attgacgaa attacgaaa caaaacgcg gaattgatg acttataag gccggagtt aacgagctg aagaataaa ttgacggcg ttcaaagcg aatgaggct gaaattaag aacttgaac gctcgtctg attgagaaa gacaaagtc attgataaa ttgaaggag gatcttaag ttggaacgg gacgaaaaa attgacttg attaacgaa catgagaaa tatcgacaa gaaacgcaa aagcagcgt gatttatgg acgcaggaa acaaacaaa ttgcgtaaa gagctagac aatatcaac gagatcgtt aaaatgaac cagaaggga gccgaagaa aatctcaaa gtgcgactc gaagaggag aaaatgttg attttgaat gagcaagat agtgatagg gaagcctac cagaagctt ttacaagag taccattgc ttggaacaa cactgcgaa gaactagag aagcagttg aacaatcag agccaaaac caaagttcc caccgaaga aatgtgagc gatcttagc agcattgac agttttgtt atcaacagc gatattgct gaagaccac ggttacggt tccgtgaga tcgaccaca acttcgtcg tcacaaaga gaaaaactt gagaatatc gactggaaa tgtgaagga aatgtagat agtcaaaca ccaagtaca agtagcagt gacaccaaa catgaatcc gaaaacaat aaagttgac gtcggtttg gtccttaaa ctccagcac aaacttgcc gaagttgag cgtgaaaaa ctgcgaatg cagaaacgc ctagacgag ttggacatg tctccacga accgaaaag gctgaaaac gcggcacga gattccatt aaaatctcc gaacttgaa ctagttaac agcaaccta aaggcgcaa ctgatggaa ctgcaaaac agcattatt gaaggaact ggcagttcg aaaatacac gaacagatg caaatatta cacagcgaa ctggaacga aaaactgag gaaataatc caactcaag agtgtcttg gccgaccag actaacaac atgaaatca ctcgtgaac tcaaatgca cgattagga gcgtacatt aatgaagac ggagagtta gcactggct tacgaaacc caaaaaaca atcaataag caactcgaa ttggaatta caagacgag aaatcaaaa tacaaagct cacgaacgg gaatacaag ctagagatt gaaaaatta agggaagat aatgaacgc caacaacga cttttatcc gcaaatctc accaacacg ccacaaaca caaagcgaa gcttttatg caacacgaa attacaaga ttaactata gaaaatctt gacttacac gacaaaaac gatactttg accgagaca gtccgaaaa cttaagaaa caaatcaag ttaatggcc aagaaactt aaagaagca gggctggat atggacgat aattttaac gaaattacc gacaacgaa cctaacaaa acgaatgta aacaagcac actcgggcc ctgccgtcg atcagaaag aaagaaaga gattattta ggcatgttc tcgtattct agtggggag gaaaatacg atcatgcgg caacttgtt attgatttg aagcctagg actgcggtc actttgctt cctggcttg ccagcttac atagttttc atgtgtata agacacacg gattacgtc aacgacgaa gataaagtg aaagccttg ctctccgcg tttacaaac tctgtaaaa aaagtgata aagaagcga cacgaagat ttcgaaacc atggcttta tggctttca aacaccttg agattagtg cacaatatg aaacagtac agcggtgac cgagctttc caggctaaa aatacgcca aaacaaaac gagcagtgt ttgcgaaat ttcgattta tcggaatat cgacaagtg ctttcagat attgctgtg tggatttac cagggactg atccgaaag tttgccgaa aaaatccaa ccacttgtc atacccgcc attctggag cacgaggaa atcccggga attagcggg aataaacct agtggcttc agagggagg agcagcagt gtagcgaca agccctgaa cccagccaa aaaccaacg accgcggtt ttactggaa ttaacaaac caccacaag attttaagt ttctatggc gttgacccg gaagtaatt tcccaaatt ttcaaacaa atattttac tttttatgc gcgacaagt ctcaataat ttgctccta cggcaagag ttgtgccac tggtcgaaa ggtttccaa attcggcac aacttatcc cacttcgag atgtggaca cgagaaaaa ggccttgat gaggcttca atccagtca actttacag cccataatt caagcggcc catttatta caagcgcgt aaaaccgaa gaagacgtt gcaagtgtg tgtgaaatg tgtagcgct ttaacgcca ctacaaatt tgtaaaata ctgaatttg tataaacct gtagatgaa tttgaacaa cacgtgccc ccgtcattt attagcaaa gttaaagcc aagttacag gagcggaat cccacttcg gaacaacaa acgctgctc atggacgta aagtatcac ttcccagtc cgattcccg ttcaatcct tccgtcata tgtttggaa gacattgaa attccagaa gttttaaaa ttgcctcag ttggaaaaa ctataa
- >TC009935 RA
gtacctaac cggaattca ttcctagtg atccccaga gggcgctag caattcttt tgagcgcca aatcgccaa atccatact tttacacat gactcctag tgatcgaaa taagccacg cagtgatta gtgcgcatg gtgcgccgt gtaggtgca ggttgtgtg ttttactac cattataat acacgagag ggttttcta tttgttggc gggttgtgc tggctccgg attattcgt ttgattact ttttttttg atgatttat gaacaagat cagtgaaaa cggtgcaaa atgactacc ttggagcta tatacaaag ggtgccaag gtatggata cctcaccct gaaaaagta tgggaaggc gctgaactt gaagaagac tacaaaacg aacaaaaag tcacttgaa gtagtaaca gaaaacaac gaaaagaaa acccttacg atcaaatca gaatcggat ttaccctgt ctgaggaac ccttcaatt ttagtcggg gaaaatgac cttacatcc ctctcctac ttacacgag cccgccgta ctttacaat ttacaagta cggttctgc caaaatcga tacatttac acctattgt gggattgtt ttggtggca attaatccc tacgatgat cttcctatt tatgacgtc gacaccata caaacgtat agaggtcaa gcgatgggt gacctagat cctcacata ttcgccgta gccgaagaa gcgtacacg aaactagaa agagagcaa agggaccaa agcataatc gtatcagga gaaagcgga gccggtaaa acggtttcc gcgaaatac gcaatgaga tacttcgcg actgtagga ggaagtgcg accgaaacc caagtcgaa aaaaaagtc ctagctagt agtcctatc atggaagcc atcggaaac gcgaaaact acaagaaac gataacagt tcgcgattt ggaaagttt atagaactc cagtttaat aagcaattt catattagc ggagcgtcg atgagaacg tacctcttg gaaaaatcg cgagtcgtc tttcaggct cccgatgaa cggaattac catattttc taccaattg tgttccgcg agagaccaa tttccacac ttgcatctc gctcaccaa gatagtttt tattattta aatcaagga gaaagtcca aatgtggac ggagtcgac gatttacaa accttcgaa gagacgctc tatgcgttg aatctttta gggtttaag aaatcggat ctggacgat atgtttaaa attatagcc tccgtcttg catttggga aatatcaag tttgttgaa tcgatcatt tccacggaa aacgaacaa gaccaggag ggttgtagc atactgggc actgactca catttgaaa atcttagcg gaattactc gagatcgat tcaaaagaa atgcagcag tggctctgt acccgaaaa atcgtctct atgagagag gtcttcctg aaaccaatg tccgtcgaa gacgcgcaa gctgcccgt gatgcttta gcaaaacac atttatgca gaattgttc aactggatc gtgttagtt ataaacaaa gctttagaa tccgacatt cctagacat aaatttatc ggagtcctc gatatctac ggtttcgaa accttcgaa accaattcc tttgaacaa ttttgtatc aattacgcc aacgaaaaa ctccaacaa caattcaac ctccacgtg tttaaactg gagcaggag gaatacatt aaagaagga attgaatgg aaaatgatt gatttctac gacaaccaa ccctgcatc gacctcatc gaaacaaaa ttagggatt ttagatttg ctagacgag gagtgtcgg atgccgcga ggtactgac agttcctgg acggagaaa ctgtactca aagtgcgca aagtattcc cacttcggc aaggcgcgt ttcggcact tctgccttc accatcaat cactttgcc gataaagtc caatacgaa agcaacggt tttctggag aaaaaccgc gacaccgta atcgaggag caaatctcg gtggtgaaa aggagcaaa aacgatcta gtccgccgg cttttcggc gacgattct caaaaattg gccgttccg ggagccaaa ctcaaagtg atctcggcc aaacctgcc ccaacctcg caaaaaacc cacaagaaa tcggtgggg tcccagttt agagactcg cttaatatg ctaatgacc acgttgaac gccacaacc ccgcattac gtccggtgt attaagcca aacgattcg aaaaaaccg tttgagtat aacccgaaa cgggcggtg caacaactg cgagcttgt ggcgtgttg gagacgatc aggatttcg gcagccggg tttccgagt cggtggact tacgttgat ttcttctac cggtaccga gtcttgtgc aagtttaaa gatatcaaa agaaacgac atgcagctc acttgccag aatatactg aaccagtat attaaaaac aaggatatg tttcaattc gggaaaact aagattttt ttccgggcc ggacaagtc gcctatttg gagaaatta agagccgat aaactcaaa gcctgttgc atcatgatg caaaaaaca atccgggcg ttcatttgg aggaaaaag tacctccga atcaagcgc tgcactatc aatatacaa cggtatgga cggggttat ttggcccga aaactggcc gaaaatttg cggcgcgag aaagccgca aagactttg cagagatat gttcgagga tgggtgaaa cgtgtccaa tatcaaaga ctgaaggcc tgtgtgact ggcattcaa aggtatgcc agagggtat ttagcgaga cggcggtac atgcaatta cggtacaac gccaaagct ctggtaata cagcgatat gtaagaggg tatttagca cggagaagt gccctggct tacaagcgg aaaatcgtc atatgccaa gctgcgatt cgccgcttt ttggctaga agattatac aagaaactc agaatcgaa gcaagatca atcgaacac gtcaagaaa cttaacaaa ggcttggag aacaagatc atcagtttg cagcaaaag attgacgaa attacgaaa caaaacgcg gaattgatg acttataag gccggagtt aacgagctg aagaataaa ttgacggcg ttcaaagcg aatgaggct gaaattaag aacttgaac gctcgtctg attgagaaa gacaaagtc attgataaa ttgaaggag gatcttaag ttggaacgg gacgaaaaa attgacttg attaacgaa catgagaaa tatcgacaa gaaacgcaa aagcagcgt gatttatgg acgcaggaa acaaacaaa ttgcgtaaa gagctagac aatatcaac gagatcgtt aaaatgaac cagaaggga gccgaagaa aatctcaaa gtgcgactc gaagaggag aaaatgttg attttgaat gagcaagat agtgatagg gaagcctac cagaagctt ttacaagag taccattgc ttggaacaa cactgcgaa gaactagag aagcagttg aacaatcag agccaaaac caaagttcc caccgaaga aatgtgagc gatcttagc agcattgac agttttgtt atcaacagc gatattgct gaagaccac ggttacggt tccgtgaga tcgaccaca acttcgtcg tcacaaaga gaaaaactt gagaatatc gactggaaa tgtgaagga aatgtagat agtcaaaca ccaagtaca agtagcagt gacaccaaa catgaatcc gaaaacaat aaagttgac gtcggtttg gtccttaaa ctccagcac aaacttgcc gaagttgag cgtgaaaaa ctgcgaatg cagaaacgc ctagacgag ttggacatg tctccacga accgaaaag gctgaaaac gcggcacga gattccatt aaaatctcc gaacttgaa ctagttaac agcaaccta aaggcgcaa ctgatggaa ctgcaaaac agcattatt gaaggaact ggcagttcg aaaatacac gaacagatg caaatatta cacagcgaa ctggaacga aaaactgag gaaataatc caactcaag agtgtcttg gccgaccag actaacaac atgaaatca ctcgtgaac tcaaatgca cgattagga gcgtacatt aatgaagac ggagagtta gcactggct tacgaaacc caaaaaaca atcaataag caactcgaa ttggaatta caagacgag aaatcaaaa tacaaagct cacgaacgg gaatacaag ctagagatt gaaaaatta agggaagat aatgaacgc caacaacga cttttatcc gcaaatctc accaacacg ccacaaaca caaagcgaa gcttttatg caacacgaa attacaaga ttaactata gaaaatctt gacttacac gacaaaaac gatactttg accgagaca gtccgaaaa cttaagaaa caaatcaag ttaatggcc aagaaactt aaagaagca gggctggat atggacgat aattttaac gaaattacc gacaacgaa cctaacaaa acgaatgta aacaagcac actcgggcc ctgccgtcg atcagaaag aaagaaaga gattattta ggcatgttc tcgtattct agtggggag gaaaatacg atcatgcgg caacttgtt attgatttg aagcctagg actgcggtc actttgctt cctggcttg ccagcttac atagttttc atgtgtata agacacacg gattacgtc aacgacgaa gataaagtg aaagccttg ctctccgcg tttacaaac tctgtaaaa aaagtgata aagaagcga cacgaagat ttcgaaacc atggcttta tggctttca aacaccttg agattagtg cacaatatg aaacagtac agcggtgac cgagctttc caggctaaa aatacgcca aaacaaaac gagcagtgt ttgcgaaat ttcgattta tcggaatat cgacaagtg ctttcagat attgctgtg tggatttac cagggactg atccgaaag tttgccgaa aaaatccaa ccacttgtc atacccgcc attctggag cacgaggaa atcccggga attagcggg aataaacct agtggcttc agagggagg agcagcagt gtagcgaca agccctgaa cccagccaa aaaccaacg accgcggtt ttactggaa ttaacaaac caccacaag attttaagt ttctatggc gttgacccg gaagtaatt tcccaaatt ttcaaacaa atattttac tttttatgc gcgacaagt ctcaataat ttgctccta cggcaagag ttgtgccac tggtcgaaa ggtttccaa attcggcac aacttatcc cacttcgag atgtggaca cgagaaaaa ggccttgat gaggcttca atccagtca actttacag cccataatt caagcggcc catttatta caagcgcgt aaaaccgaa gaagacgtt gcaagtgtg tgtgaaatg tgtagcgct ttaacgcca ctacaaatt tgtaaaata ctgaatttg tataaacct gtagatgaa tttgaacaa cacgtgccc ccgtcattt attagcaaa gttaaagcc aagttacag gagcggaat cccacttcg gaacaacaa acgctgctc atggacgta aagtatcac ttcccagtc cgattcccg ttcaatcct tccgtcata tgtttggaa gacattgaa attccagaa gttttaaaa ttgcctcag ttggaaaaa ctataaata attctagca aatatttta accaaactc acgtttgca gtgtaaata atgtcaact tagtcac
