| General information | |||
| TC015405 | 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: 442708..452354 | |
| 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 | TC015405 | ||
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_05812 | 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: 0.0% (includes death as pupa, adult)
| Many background defects. None of them is really penetrant. |
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: -1.0% (includes death as larva, prepupa, pupa)
Lethalities 22 days after larval injection: -1.0% (includes death as larva, prepupa, pupa, adult)
- >TC015405 RA
MDPENKPKT NEKLDSTLK KFLSAQPFD SDDITESAK QIFQELNQN YDHFKLVTI SDDLDEILP KKRDPKTPI YAILEKLKQ RWVVLCLWG DKAYYKDSS GEKIPHRIE KKFDTLTIF ENFDQDHDA DAVENYGPF SLRNLQILL KLGQEIGEK FPSQKFCQL ADVDEEKVN IIVKNGQEF AQKNDFKNA LRQFIETLP LDLGARMLK YKQILEEEL DKDETLVDH ERIKLARLE YLTDENVTK LQENYKVND SSVEIEVVK QFPEEMDKL GQIFATLDH ITDQKLREN ALEQLAILL NLDANKLKS FYASKGPKT PLKKPQLTL DDISKIVDN LPKLDKTTK DAETTPFDQ LLDQVTLGQ DLTPSSPAQ FTPNKSQLE QNYYQVKEK YQLWRGSTL HDINTWARS QKGKPGNFC EIVAVCDRV FALLTGGHR LRDTQILAV LLFNDVEKR GRLCQVQTG EGKTTIVSI LAVVGALQG HLIDVVTTS PVLASDGVD KNRDFYAVF GLTVGTTDT PGGYTADVL YGSIASFQF DYLRESFEG LNVRNDRRF GLVILDEVD SLLVDNGGH IAKIATPYP GMESLRYVF IKIWAELAK AEERFIEEN QSKLESIVT NDSAEQLFD EFWAQDERQ VIRDLIKSA MPATSPLIP QHLSHYVAR NLDDWIESA LVAKYDCHE LEQYRIVNG VVTPVDNLN TGVTLTNTV WSKGLQQFL QLKHNLHLT CSTLSSIFI SNFGYIQKY AKIFGVTGT LGSTTEQDL LSHVYNVTF AKIPTHKPK SFTEEPPIV TKDNSWDLE VTSEIMRKI DEKRAVLVI FATERDLNT VGRNLEIIR HDDFEIRYI RHEGEVKDA SLVVKASSV VLATNLAGR GCDFKTDQS VEERGGLHV IVAFLPANQ RVEDQAFGR TSRNGNRGS GQLIVRESE ASGGGLKQT RDLQERGRL KHVRDVLIK EIVFKEDLI GRFADLYLK LKRAKVLEG EFCYFLQDL KEFWAFWLD SRNFHMITQ SVDEVFGEF LGEAEYIIK GEVKHNPFY SIALADHYL ENNKKAKAR SELLKVVSS GANHTELLA GAHLKMFEL AIASGNQTW HKIRKALAK VLFIDVQKN TLYKKNALK QLGHANAAI TTEIDFIES WLQSPDFER ILPKDNFLK QVNSRLYCL KIQKGNIEE LVKQIEEAG DDGVDFGGK AQSTVGNYR KMDKSSKEI VGETELSEL QAVGIDALY TLRTVKDVP DYVIHAAQG QIGAGIAAL AAGLFFPPI FPIMSPLGV TLISEGIID IVFALFEDN PEFKDFMKG KAVSYGIAL VTFGLSAIA QSIKIITKA LSFFRKLAT FLRKSSYLK GLSAKLAKV VDKLANKME RLLDVAKFN KLTRSQQLS QLKELQKSG KAFDHLGGL SKLQSLEKL QKLGKLKEL TRSQLLVQF AKEVAWKTA EEGGKRLLE KFVIKPFLG RLYGDVIEK IKAFLVKSL KQSIQDNAN LREKLRTTD KKQIDETLE RFIKEDVVI EKLRDYLFD SLGKFKSTK FHVLIHGVK MAVNVGKMV HICTKFAAN LEKQLKGGD ADNNVDKII DKICTKLVD EVLTICWDV LKDNVKFVW KHKDVFASK PTDLVLKPA EKTGNSWKV FGLEPGASL DEVKTHYRK MALVLHPDK NLNDPTATE KFQEFQNAY EDILKTF
- >TC015405 RA
atggatcca gaaaacaag ccaaaaacc aacgaaaaa cttgactca actctcaaa aaattcctc tcggctcaa cctttcgac tcagacgac atcaccgaa tccgccaaa caaatcttc caagaattg aaccaaaac tacgaccac ttcaaacta gtgacaatc tccgacgat ttggacgaa attctccca aaaaaacgc gaccccaaa accccaatt tacgcaatt ttggaaaaa ttgaaacaa cgatgggtc gttttgtgc ctttggggg gacaaagcc tactacaag gactcatca ggcgaaaaa attccccat cgaattgaa aaaaaattc gacactttg acaattttc gaaaatttt gaccaagac cacgacgct gacgcggta gagaattac gggcctttc agtcttcgg aatttacaa attttgcta aagttgggc caagaaatc ggcgaaaaa ttcccaagc caaaaattt tgccaattg gctgatgtt gacgaagag aaagtcaat ataattgtc aaaaatggg caagaattt gctcaaaaa aatgatttt aaaaacgcc ctacgacag tttattgag acactaccg ctagatttg ggggcacgt atgctaaag tacaagcaa atccttgag gaagagctg gataaggac gaaactttg gttgaccac gagcggatc aagttggct cgtctggag tacctaacc gacgaaaat gtcactaaa ttgcaagaa aattacaaa gtcaatgat tcaagtgtt gaaatcgaa gtcgtcaaa caatttccg gaggaaatg gacaagttg ggccaaatt tttgcaact cttgaccac atcactgat cagaaactg cgtgaaaac gcattagaa cagctcgcg attttgcta aatttagac gccaacaag ctcaagtca ttttatgcg tccaaaggc ccgaaaacg ccactcaaa aaaccgcaa ctaaccctt gacgatatc agcaaaatt gtcgacaat ttgccaaaa cttgacaaa acgaccaag gacgcggaa accaccccc tttgaccaa cttttggat caagtgacc cttggccaa gacctgacc ccttcatcc cccgcccag ttcaccccc aacaagagt caactcgaa caaaactac taccaggtc aaggaaaaa taccaactt tggcgaggt agcacttta cacgacatt aacacttgg gctaggagt caaaagggc aaacctggc aatttttgc gaaattgtg gcggtttgt gaccgcgtt ttcgccctt ttgacaggt gggcaccgc ctacgtgat acccagatt ttggccgtt ttgctcttc aatgacgtt gaaaaacgc ggtcgcctg tgccaggtc caaacaggt gaaggcaaa acaacaata gtgtcaatt ctggcagtt gtgggggcc ttgcagggc cacttgatt gacgtggtc accactagt ccagtgttg gcaagtgat ggtgttgac aaaaatcgg gatttttac gccgttttt gggctcacg gttggcacc actgacacg ccgggtggc tacactgcc gatgtgctg tacgggagt attgccagt ttccagttt gattatttg agggaaagt tttgagggg ttaaatgtt cgaaatgat cggcggttt ggtctagtt atcttggac gaggttgat agtcttttg gttgacaat ggcggacat attgccaaa attgcaacc ccgtacccg ggaatggag agtctaagg tacgttttt atcaaaatt tgggctgag ttggccaag gctgaggaa cgttttatt gaggaaaac cagtcaaaa ctagagtca attgtgaca aatgacagt gcggagcaa ttgtttgac gagttttgg gcgcaagac gaacgccaa gtcattcgg gacctaatc aagagtgcc atgccggca accagccca ctaatcccc cagcacttg tcacactat gttgcgcgc aatttggac gattggatc gagagtgcc ttggtcgcc aagtacgat tgtcacgag ttggaacaa tatcgcatc gtcaacggt gttgtgact cctgttgac aacctaaac accggcgtg accttgact aatacagtg tggtccaaa ggactccaa cagttcctc caattgaag cacaatctt catctgacc tgttctacc ctaagtagc atctttatt tccaacttt ggttacatc cagaagtat gccaaaatt tttggggtc actggaacc ttgggctcc accaccgag caagacctg ctatcccac gtctataat gtcactttt gccaaaata ccaacccac aaacccaaa agcttcacc gaagaaccg ccaattgtc acaaaagac aatagttgg gatttggaa gtaacgtcc gaaatcatg cgaaaaatt gacgaaaag cgggccgtt ttggtgatt tttgccacc gaacgtgac cttaacacc gttggacgt aaccttgaa atcatccgc cacgatgac ttcgaaatc cgctacatt cggcacgaa ggtgaagtc aaggacgcc agcctagta gtaaaggcc tctagcgtt gtcttggct acgaacctc gcggggcgt ggctgcgac ttcaaaacg gaccaatcg gtggaggag aggggcggt ttgcacgtc attgtggcc ttcctaccg gccaatcag cgtgtggaa gaccaggcc ttcgggcgg acgtcacgc aacggaaat aggggaagt gggcaattg atcgtgcgc gagagtgag gcgagtggt gggggtttg aaacaaaca cgtgatcta caagagagg gggcgacta aaacacgtt cgtgacgtt ttaatcaaa gaaatcgtc ttcaaggag gacttgatc gggcgtttt gctgacttg tacctgaaa ctgaaacgt gccaaagtg ctggaaggg gagttttgc tactttttg caggatttg aaggagttt tgggccttt tggttggat tcccgcaac ttccacatg atcacacaa agtgtggac gaagttttt ggcgagttt ttgggcgaa gctgagtat attattaag ggagaagtt aaacataat cccttttat tccatcgca ttggctgac cattactta gaaaacaat aaaaaagct aaggctaga agtgagttg ctcaaggtg gtgtcaagt ggtgccaac catacggaa ttgttggct ggggcccac ttgaaaatg ttcgaactg gcgatagct agtgggaac cagacgtgg cacaagatc aggaaggcg ttggctaag gttttgttt attgacgtt cagaagaac acactgtat aagaaaaac gccttaaaa cagttgggg cacgctaac gcggccata actaccgag attgatttt atcgaaagt tggctacaa agtcctgat tttgagcga atccttcca aaggataat tttctaaaa caagtgaat tcaaggttg tattgtttg aaaattcag aaagggaat attgaggag ttggtgaaa cagatcgaa gaggctggg gacgacggg gttgatttt ggggggaag gcgcaatcg acggttggg aattatcga aagatggac aaaagttct aaagaaatc gtaggtgag acagagttg agtgaactc caagccgtt ggtatcgac gctctttac acccttcga actgtcaaa gacgtccct gattacgtc atccatgcg gctcaaggt caaatcggt gcaggaatt gcagctttg gccgcagga ctttttttc ccgcccatt tttccaatc atgagccca cttggtgta accctaatt tccgaaggg ataatagac attgttttc gcccttttc gaggacaat cccgagttc aaggacttt atgaagggc aaagcggtg tcatacggc attgccttg gttactttc ggcctcagc gccatcgcc caatcaatc aaaatcata accaaagct ttgagcttt ttccgaaag ttggcaact ttcctacgc aaaagctcc tacttgaag ggcttaagt gccaagttg gccaaagtt gtggacaag ttagcgaac aaaatggag cgcctacta gatgtcgcc aaatttaac aaactgaca aggagtcaa caactaagc caactgaaa gagttgcaa aaaagcggc aaagctttc gaccatttg ggcggttta tccaagtta cagagtttg gaaaagttg caaaaactg ggcaaattg aaggaactg acccgcagc caactcctg gtgcagttc gccaaagag gtggcgtgg aaaacggca gaagaaggc ggcaaacgt ttattggaa aaatttgtt attaaaccg tttctgggc cgactttac ggcgacgtg atcgaaaaa atcaaggcg tttctggtc aaaagtctg aaacaatcg attcaagac aacgccaac ttgcgcgaa aaactccga acgactgat aaaaaacaa attgacgaa actttggag cgctttatt aaagaagac gtcgttatt gaaaaattg cgtgactat ctttttgac agtttggga aaattcaaa agtacaaag tttcacgtt ttgatccac ggagtcaag atggccgtg aatgtgggc aagatggtt cacatctgt acgaaattt gcggcaaat cttgaaaaa caactgaaa ggtggggac gctgacaac aacgttgat aagataatt gataaaatt tgcacaaag ttggtcgat gaggtcttg acgatttgt tgggacgtc ctgaaggat aatgtcaaa tttgtatgg aaacataag gacgttttt gcaagtaaa ccgactgat cttgtcctg aaaccggct gaaaaaact ggaaattcc tggaaagtt tttgggctt gaaccagga gcgtctctt gatgaggtt aagacgcat tacaggaag atggcgttg gttttgcac cccgataag aatttgaac gatccgacg gccactgaa aagtttcag gagtttcag aatgcttac gaagacatt ctgaagaca ttttga
- >TC015405 RA
caggtagta aaatctaga ggcacgtgt gagtgcaaa agttaattt tttcactga cctgacagt agaaaagcc ttggctttg tttcccgat taataattc attgatttt catagcatt tttgcggta actgcactg agcgcacct ttgccacca actgatgaa agacataaa gcccaagag gatatttat agttcggac gccatttcc gagcgctta tttcgacac cgtgtaaac gaactttaa ttattactc ggcggaatg aaatttttt cgtttaaat tgtaaaaaa tggatccag aaaacaagc caaaaacca acgaaaaac ttgactcaa ctctcaaaa aattcctct cggctcaac ctttcgact cagacgaca tcaccgaat ccgccaaac aaatcttcc aagaattga accaaaact acgaccact tcaaactag tgacaatct ccgacgatt tggacgaaa ttctcccaa aaaaacgcg accccaaaa ccccaattt acgcaattt tggaaaaat tgaaacaac gatgggtcg ttttgtgcc tttgggggg acaaagcct actacaagg actcatcag gcgaaaaaa ttccccatc gaattgaaa aaaaattcg acactttga caattttcg aaaattttg accaagacc acgacgctg acgcggtag agaattacg ggcctttca gtcttcgga atttacaaa ttttgctaa agttgggcc aagaaatcg gcgaaaaat tcccaagcc aaaaatttt gccaattgg ctgatgttg acgaagaga aagtcaata taattgtca aaaatgggc aagaatttg ctcaaaaaa atgatttta aaaacgccc tacgacagt ttattgaga cactaccgc tagatttgg gggcacgta tgctaaagt acaagcaaa tccttgagg aagagctgg ataaggacg aaactttgg ttgaccacg agcggatca agttggctc gtctggagt acctaaccg acgaaaatg tcactaaat tgcaagaaa attacaaag tcaatgatt caagtgttg aaatcgaag tcgtcaaac aatttccgg aggaaatgg acaagttgg gccaaattt ttgcaactc ttgaccaca tcactgatc agaaactgc gtgaaaacg cattagaac agctcgcga ttttgctaa atttagacg ccaacaagc tcaagtcat tttatgcgt ccaaaggcc cgaaaacgc cactcaaaa aaccgcaac taacccttg acgatatca gcaaaattg tcgacaatt tgccaaaac ttgacaaaa cgaccaagg acgcggaaa ccaccccct ttgaccaac ttttggatc aagtgaccc ttggccaag acctgaccc cttcatccc ccgcccagt tcaccccca acaagagtc aactcgaac aaaactact accaggtca aggaaaaat accaacttt ggcgaggta gcactttac acgacatta acacttggg ctaggagtc aaaagggca aacctggca atttttgcg aaattgtgg cggtttgtg accgcgttt tcgcccttt tgacaggtg ggcaccgcc tacgtgata cccagattt tggccgttt tgctcttca atgacgttg aaaaacgcg gtcgcctgt gccaggtcc aaacaggtg aaggcaaaa caacaatag tgtcaattc tggcagttg tgggggcct tgcagggcc acttgattg acgtggtca ccactagtc cagtgttgg caagtgatg gtgttgaca aaaatcggg atttttacg ccgtttttg ggctcacgg ttggcacca ctgacacgc cgggtggct acactgccg atgtgctgt acgggagta ttgccagtt tccagtttg attatttga gggaaagtt ttgaggggt taaatgttc gaaatgatc ggcggtttg gtctagtta tcttggacg aggttgata gtcttttgg ttgacaatg gcggacata ttgccaaaa ttgcaaccc cgtacccgg gaatggaga gtctaaggt acgttttta tcaaaattt gggctgagt tggccaagg ctgaggaac gttttattg aggaaaacc agtcaaaac tagagtcaa ttgtgacaa atgacagtg cggagcaat tgtttgacg agttttggg cgcaagacg aacgccaag tcattcggg acctaatca agagtgcca tgccggcaa ccagcccac taatccccc agcacttgt cacactatg ttgcgcgca atttggacg attggatcg agagtgcct tggtcgcca agtacgatt gtcacgagt tggaacaat atcgcatcg tcaacggtg ttgtgactc ctgttgaca acctaaaca ccggcgtga ccttgacta atacagtgt ggtccaaag gactccaac agttcctcc aattgaagc acaatcttc atctgacct gttctaccc taagtagca tctttattt ccaactttg gttacatcc agaagtatg ccaaaattt ttggggtca ctggaacct tgggctcca ccaccgagc aagacctgc tatcccacg tctataatg tcacttttg ccaaaatac caacccaca aacccaaaa gcttcaccg aagaaccgc caattgtca caaaagaca atagttggg atttggaag taacgtccg aaatcatgc gaaaaattg acgaaaagc gggccgttt tggtgattt ttgccaccg aacgtgacc ttaacaccg ttggacgta accttgaaa tcatccgcc acgatgact tcgaaatcc gctacattc ggcacgaag gtgaagtca aggacgcca gcctagtag taaaggcct ctagcgttg tcttggcta cgaacctcg cggggcgtg gctgcgact tcaaaacgg accaatcgg tggaggaga ggggcggtt tgcacgtca ttgtggcct tcctaccgg ccaatcagc gtgtggaag accaggcct tcgggcgga cgtcacgca acggaaata ggggaagtg ggcaattga tcgtgcgcg agagtgagg cgagtggtg ggggtttga aacaaacac gtgatctac aagagaggg ggcgactaa aacacgttc gtgacgttt taatcaaag aaatcgtct tcaaggagg acttgatcg ggcgttttg ctgacttgt acctgaaac tgaaacgtg ccaaagtgc tggaagggg agttttgct actttttgc aggatttga aggagtttt gggcctttt ggttggatt cccgcaact tccacatga tcacacaaa gtgtggacg aagtttttg gcgagtttt tgggcgaag ctgagtata ttattaagg gagaagtta aacataatc ccttttatt ccatcgcat tggctgacc attacttag aaaacaata aaaaagcta aggctagaa gtgagttgc tcaaggtgg tgtcaagtg gtgccaacc atacggaat tgttggctg gggcccact tgaaaatgt tcgaactgg cgatagcta gtgggaacc agacgtggc acaagatca ggaaggcgt tggctaagg ttttgttta ttgacgttc agaagaaca cactgtata agaaaaacg ccttaaaac agttggggc acgctaacg cggccataa ctaccgaga ttgatttta tcgaaagtt ggctacaaa gtcctgatt ttgagcgaa tccttccaa aggataatt ttctaaaac aagtgaatt caaggttgt attgtttga aaattcaga aagggaata ttgaggagt tggtgaaac agatcgaag aggctgggg acgacgggg ttgattttg gggggaagg cgcaatcga cggttggga attatcgaa agatggaca aaagttcta aagaaatcg taggtgaga cagagttga gtgaactcc aagccgttg gtatcgacg ctctttaca cccttcgaa ctgtcaaag acgtccctg attacgtca tccatgcgg ctcaaggtc aaatcggtg caggaattg cagctttgg ccgcaggac tttttttcc cgcccattt ttccaatca tgagcccac ttggtgtaa ccctaattt ccgaaggga taatagaca ttgttttcg cccttttcg aggacaatc ccgagttca aggacttta tgaagggca aagcggtgt catacggca ttgccttgg ttactttcg gcctcagcg ccatcgccc aatcaatca aaatcataa ccaaagctt tgagctttt tccgaaagt tggcaactt tcctacgca aaagctcct acttgaagg gcttaagtg ccaagttgg ccaaagttg tggacaagt tagcgaaca aaatggagc gcctactag atgtcgcca aatttaaca aactgacaa ggagtcaac aactaagcc aactgaaag agttgcaaa aaagcggca aagctttcg accatttgg gcggtttat ccaagttac agagtttgg aaaagttgc aaaaactgg gcaaattga aggaactga cccgcagcc aactcctgg tgcagttcg ccaaagagg tggcgtgga aaacggcag aagaaggcg gcaaacgtt tattggaaa aatttgtta ttaaaccgt ttctgggcc gactttacg gcgacgtga tcgaaaaaa tcaaggcgt ttctggtca aaagtctga aacaatcga ttcaagaca acgccaact tgcgcgaaa aactccgaa cgactgata aaaaacaaa ttgacgaaa ctttggagc gctttatta aagaagacg tcgttattg aaaaattgc gtgactatc tttttgaca gtttgggaa aattcaaaa gtacaaagt ttcacgttt tgatccacg gagtcaaga tggccgtga atgtgggca agatggttc acatctgta cgaaatttg cggcaaatc ttgaaaaac aactgaaag gtggggacg ctgacaaca acgttgata agataattg ataaaattt gcacaaagt tggtcgatg aggtcttga cgatttgtt gggacgtcc tgaaggata atgtcaaat ttgtatgga aacataagg acgtttttg caagtaaac cgactgatc ttgtcctga aaccggctg aaaaaactg gaaattcct ggaaagttt ttgggcttg aaccaggag cgtctcttg atgaggtta agacgcatt acaggaaga tggcgttgg ttttgcacc ccgataaga atttgaacg atccgacgg ccactgaaa agtttcagg agtttcaga atgcttacg aagacattc tgaagacat tttgatttg tgggaaaat agttttttt tggtgattt caacaagcg acgaaccct gtaatttac gtaatcaat aaagtgttc aagtcagt
