| General information | |||
| TC005855 | 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_007423.3: 6382217..6410632 | |
| 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. | |||
| Closest fly homologs |
|
TC005855 | |
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_00945 | 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: 3 (3 dpi)adult nutritional state bad (cachectic) - Number of animals showing the phenotype: 2 (11 dpi)
adult nutritional state potentially bad (cachectic) - Number of animals showing the phenotype: 2 (11 dpi)
affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): 50% - 80%
embryo/egg without visible larval cuticle in cuticle preparation - Penetrance (percentage of animals/eggs on slide): 50% - 80%
affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): 50% - 80%
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)
- >TC005855 RA
MEGKAGHKF VGRGAHKGK GIAVFTSGG DSQGMNAAV RAVVRMGIY LGCKVYFIK EGYQGMVDG GKYIVEANW SSVSSIIHK GGTVIGSAR CQDFRERAG RLKAAKNLV ENGITNLVV IGGDGSLTG ADLFRQEWS SLLEELATT GQITKEQQQ KYRTLQIAG LVGSIDNDF CGTDMTIGT DSALHRIIE ACDAIVSTA YSHQRTFIM EVMGRHCGY LALVAALTS EADYVFIPE SPPPDDWQK RLCDKLEQS TKDRNEGKL FKSCLGKPP MIMMNDFNW LLSSFFFQG APPRPLAWN PTLRQDCYL ALISALATE ADYAFIPEN PADVNWHEK LCHKLSQER HMGQRLNII IVAEGAIDR NGDPITAEM IKKVVVDNL QQDTRITVL GHVQRGGNP SAFDRILGC RMGAEAVMA LMEADENTE PCVISLDGN QAIRLPLMD CVKRTKAVS QAMADKKFD LAVELRGKS FMRNLETYK LLTRLKPPK DAFDEQGHG REGFTVAVM HIGAPACGM NAAVRSFVR NFIYRGDTV LGIQEGVEG LIAGNIKQM DWAEVAGWV GIGGANLGT KRTLPGNRM GLVAARLAE FKIQGLLII GGFEAFAAA VQFLDSRKD FPALNIPIV VIPGTISNN VPGTEFSLG CDTALNEIT EICDRIRQS AQGTKRRVF IIETMGGYC GYLATLAGL AGGADAAYI YEEKFTIKD LQQDVYHMA TKMSEGVQR GLILRNEKA SDNYNTDFI YRLYCEEGK GLFSARMNV LGHMQQGGS PTPFDRNVG TKMASKAVE WMINQLKSN MSSDGSVSC SSQDSVVLL GIVKRQYKF TPVAELKEQ TNFDLRMPK EQWWLKLRP LLRILAKHD STYEEEGMY MTIEDRSFI N
- >TC005855 RB
MEGKAGHKF VGRGAHKGK GIAVFTSGG DSQGMNAAV RAVVRMGIY LGCKVYFIK EGYQGMVDG GKYIVEANW SSVSSIIHK GGTVIGSAR CQDFRERAG RLKAAKNLV ENGITNLVV IGGDGSLTG ADLFRQEWS SLLEELATT GQITKEQQQ KYRTLQIAG LVGSIDNDF CGTDMTIGT DSALHRIIE ACDAIVSTA YSHQRTFIM EVMGRHCGY LALVAALTS EADYVFIPE SPPPDDWQK RLCDKLEQS TKDRNEGKL FKSCLGKPP MIMMNDFNW LLSSFFFQG APPRPLAWN PTLRQDCYL ALISALATE ADYAFIPEN PADVNWHEK LCHKLSQER HMGQRLNII IVAEGAIDR NGDPITAEM IKKVVVDNL QQDTRITVL GHVQRGGNP SAFDRILGC RMGAEAVMA LMEADENTE PCVISLDGN QAIRLPLMD CVKRTKAVS QAMADKKFD LAVELRGKS FMRNLETYK LLTRLKPPK DAFDEQGHG REGFTVAVM HIGAPACGM NAAVRSFVR NFIYRGDTV LGIQEGVEG LIAGNIKQM DWAEVAGWV GIGGANLGT KRTLPGNRM GLVAARLAE FKIQGLLII GGFEAFAAA VQFLDSRKD FPALNIPIV VIPGTISNN VPGTEFSLG CDTALNEIT EICDRIRQS AQGTKRRVF IIETMGGYC GYLATLAGL AGGADAAYI YEEKFTIKD LQQDVYHMA TKMSEGVQR GLILRNEKA SDNYNTDFI YRLYCEEGK GLFSARMNV LGHMQQGGS PTPFDRNVG TKMASKAVE WMINQLKSN MSSDGSVSC SSQDSVVLL GIVKRQYKF TPVAELKEQ TNFDLRMPK EQWWLKLRP LLRILAKHD STYEEEGMY MTIEDRSFI N
- >TC005855 RA
atggaagga aaggccggg cataagttt gtggggcgc ggagcccat aagggcaaa ggcatcgct gtgtttacc agtggagga gattcacag gggatgaat gcggctgtc agggctgtg gtcaggatg gggatttat ttggggtgc aaagtctat tttatcaag gaaggctac caaggcatg gttgatggc ggcaagtac atcgttgag gccaattgg tcctccgta agctctatt attcacaaa ggaggcact gtgattgga agcgctcgt tgccaagat ttccgtgaa cgcgctgga cgtctcaaa gctgcaaaa aacctcgta gaaaacggc attaccaac cttgtagta attggaggt gatggtagt ttaacaggt gctgacctc ttccgtcaa gaatggagc tctctattg gaggaattg gccaccaca ggtcaaatc acgaaagaa cagcaacag aaataccgc acgttgcaa attgccggt cttgtgggt tcgattgat aacgatttt tgcggtact gatatgacc attgggacg gacagtgcc cttcacaga attatcgaa gcttgcgat gctatagtc agtacggct tattcgcat caaaggact tttattatg gaggttatg ggccggcat tgcgggtat ttggctctt gtggccgca ctcacttcc gaggccgat tatgtgttc ataccagaa tcaccacca ccagacgat tggcaaaaa aggctatgc gataaatta gaacagagc accaaggac aggaacgaa ggaaaactc ttcaaatcc tgccttggt aaaccaccg atgataatg atgaatgat tttaactgg cttctttct tctttcttc ttccaaggg gccccaccc cgccccttg gcgtggaac ccgacctta aggcaggat tgttattta gcattgatc agcgcgttg gcaactgaa gcagattac gcttttata ccggaaaat cctgctgac gtgaactgg cacgaaaaa ctctgtcat aagttgtct caggaacgc cacatgggc caacgtctc aatattatt atcgtggct gaaggcgct atcgaccgg aatggagac ccaatcact gcagaaatg atcaaaaaa gtcgtcgtt gataatctc caacaggac accagaatc actgttttg gggcacgtg caacgtggg ggcaacccg tctgctttt gataggatt ctgggctgt cgtatgggg gctgaggcc gtgatggcg ctaatggag gctgatgag aatacagag ccgtgtgtg atttcctta gacggcaat caggcgata aggttgccg ttaatggat tgcgtgaag cggacaaaa gccgtctca caggcaatg gctgataag aaatttgat ttggcggtg gaattacga ggaaagtca tttatgcgc aatttggag acttataag ttgttgact cgactcaag cctccaaag gatgcattt gatgagcaa ggacatgga agggaaggt ttcacagtg gccgtaatg catattgga gctcctgct tgtggcatg aatgccgct gtacgttct tttgtacga aattttatc tacagagga gacacagtt ttgggcatt caggaagga gttgaaggt cttatcgct ggcaatata aaacaaatg gactgggcc gaagtggct ggttgggtt ggcatcggg ggtgcaaat ctgggcacc aaacgcacc cttcctggc aatcgtatg ggtttagta gccgcccga ttggccgaa tttaaaatt caaggttta ctcattatt ggcggtttt gaagcattt gcagccgct gtacaattt ctagacagc cgaaaagat ttcccagct ctcaatata ccaatcgtg gtaatcccc ggaactatc agtaacaat gtaccaggg acggaattt tcactaggt tgtgacaca gccttgaac gaaatcaca gaaatttgt gatagaatc agacagtcg gctcaaggc acgaaaagg agggttttt attattgaa accatgggc gggtattgc ggatatttg gccacactt gcaggtctc gctgggggt gcggatgct gcgtacatt tacgaggaa aaattcaca attaaagac ttgcaacaa gatgtctac cacatggct acgaaaatg tccgaaggg gtacaacgt ggtttaatt ttgcgtaat gaaaaggcg tcagataac tacaatacc gactttatc tacagattg tattgtgag gaaggaaaa ggcttgttt agtgccagg atgaacgtt ttaggtcac atgcaacaa gggggttcc cccacacct tttgaccgc aacgttggt accaaaatg gcgtccaaa gcggtcgaa tggatgatc aaccaatta aaaagcaat atgagttca gacggctca gtcagttgt tctagtcaa gacagcgtc gttctgtta ggaatcgtt aaaagacaa tacaagttt acaccagtc gctgagttg aaagagcaa accaacttt gacttgcgt atgcctaaa gagcaatgg tggttgaaa ctgcgccct ctattgcgt attttggca aaacatgat tcgacatat gaggaagaa gggatgtat atgacgatt gaggatcgc agttttatt aattaa
- >TC005855 RB
atggaagga aaggccggg cataagttt gtggggcgc ggagcccat aagggcaaa ggcatcgct gtgtttacc agtggagga gattcacag gggatgaat gcggctgtc agggctgtg gtcaggatg gggatttat ttggggtgc aaagtctat tttatcaag gaaggctac caaggcatg gttgatggc ggcaagtac atcgttgag gccaattgg tcctccgta agctctatt attcacaaa ggaggcact gtgattgga agcgctcgt tgccaagat ttccgtgaa cgcgctgga cgtctcaaa gctgcaaaa aacctcgta gaaaacggc attaccaac cttgtagta attggaggt gatggtagt ttaacaggt gctgacctc ttccgtcaa gaatggagc tctctattg gaggaattg gccaccaca ggtcaaatc acgaaagaa cagcaacag aaataccgc acgttgcaa attgccggt cttgtgggt tcgattgat aacgatttt tgcggtact gatatgacc attgggacg gacagtgcc cttcacaga attatcgaa gcttgcgat gctatagtc agtacggct tattcgcat caaaggact tttattatg gaggttatg ggccggcat tgcgggtat ttggctctt gtggccgca ctcacttcc gaggccgat tatgtgttc ataccagaa tcaccacca ccagacgat tggcaaaaa aggctatgc gataaatta gaacagagc accaaggac aggaacgaa ggaaaactc ttcaaatcc tgccttggt aaaccaccg atgataatg atgaatgat tttaactgg cttctttct tctttcttc ttccaaggg gccccaccc cgccccttg gcgtggaac ccgacctta aggcaggat tgttattta gcattgatc agcgcgttg gcaactgaa gcagattac gcttttata ccggaaaat cctgctgac gtgaactgg cacgaaaaa ctctgtcat aagttgtct caggaacgc cacatgggc caacgtctc aatattatt atcgtggct gaaggcgct atcgaccgg aatggagac ccaatcact gcagaaatg atcaaaaaa gtcgtcgtt gataatctc caacaggac accagaatc actgttttg gggcacgtg caacgtggg ggcaacccg tctgctttt gataggatt ctgggctgt cgtatgggg gctgaggcc gtgatggcg ctaatggag gctgatgag aatacagag ccgtgtgtg atttcctta gacggcaat caggcgata aggttgccg ttaatggat tgcgtgaag cggacaaaa gccgtctca caggcaatg gctgataag aaatttgat ttggcggtg gaattacga ggaaagtca tttatgcgc aatttggag acttataag ttgttgact cgactcaag cctccaaag gatgcattt gatgagcaa ggacatgga agggaaggt ttcacagtg gccgtaatg catattgga gctcctgct tgtggcatg aatgccgct gtacgttct tttgtacga aattttatc tacagagga gacacagtt ttgggcatt caggaagga gttgaaggt cttatcgct ggcaatata aaacaaatg gactgggcc gaagtggct ggttgggtt ggcatcggg ggtgcaaat ctgggcacc aaacgcacc cttcctggc aatcgtatg ggtttagta gccgcccga ttggccgaa tttaaaatt caaggttta ctcattatt ggcggtttt gaagcattt gcagccgct gtacaattt ctagacagc cgaaaagat ttcccagct ctcaatata ccaatcgtg gtaatcccc ggaactatc agtaacaat gtaccaggg acggaattt tcactaggt tgtgacaca gccttgaac gaaatcaca gaaatttgt gatagaatc agacagtcg gctcaaggc acgaaaagg agggttttt attattgaa accatgggc gggtattgc ggatatttg gccacactt gcaggtctc gctgggggt gcggatgct gcgtacatt tacgaggaa aaattcaca attaaagac ttgcaacaa gatgtctac cacatggct acgaaaatg tccgaaggg gtacaacgt ggtttaatt ttgcgtaat gaaaaggcg tcagataac tacaatacc gactttatc tacagattg tattgtgag gaaggaaaa ggcttgttt agtgccagg atgaacgtt ttaggtcac atgcaacaa gggggttcc cccacacct tttgaccgc aacgttggt accaaaatg gcgtccaaa gcggtcgaa tggatgatc aaccaatta aaaagcaat atgagttca gacggctca gtcagttgt tctagtcaa gacagcgtc gttctgtta ggaatcgtt aaaagacaa tacaagttt acaccagtc gctgagttg aaagagcaa accaacttt gacttgcgt atgcctaaa gagcaatgg tggttgaaa ctgcgccct ctattgcgt attttggca aaacatgat tcgacatat gaggaagaa gggatgtat atgacgatt gaggatcgc agttttatt aattaa
- >TC005855 RA
tgtcatatt ggactagag cagaggtca ccttgaatt aaaattgat tcctttttg aaactaagt ttgaattcg cgttctgaa atcccagtg aatgtcttg aagttaaag ccccctggt cagaataag agacaagaa gtcacgatg gaaggaaag gccgggcat aagtttgtg gggcgcgga gcccataag ggcaaaggc atcgctgtg tttaccagt ggaggagat tcacagggg atgaatgcg gctgtcagg gctgtggtc aggatgggg atttatttg gggtgcaaa gtctatttt atcaaggaa ggctaccaa ggcatggtt gatggcggc aagtacatc gttgaggcc aattggtcc tccgtaagc tctattatt cacaaagga ggcactgtg attggaagc gctcgttgc caagatttc cgtgaacgc gctggacgt ctcaaagct gcaaaaaac ctcgtagaa aacggcatt accaacctt gtagtaatt ggaggtgat ggtagttta acaggtgct gacctcttc cgtcaagaa tggagctct ctattggag gaattggcc accacaggt caaatcacg aaagaacag caacagaaa taccgcacg ttgcaaatt gccggtctt gtgggttcg attgataac gatttttgc ggtactgat atgaccatt gggacggac agtgccctt cacagaatt atcgaagct tgcgatgct atagtcagt acggcttat tcgcatcaa aggactttt attatggag gttatgggc cggcattgc gggtatttg gctcttgtg gccgcactc acttccgag gccgattat gtgttcata ccagaatca ccaccacca gacgattgg caaaaaagg ctatgcgat aaattagaa cagagcacc aaggacagg aacgaagga aaactcttc aaatcctgc cttggtaaa ccaccgatg ataatgatg aatgatttt aactggctt ctttcttct ttcttcttc caaggggcc ccaccccgc cccttggcg tggaacccg accttaagg caggattgt tatttagca ttgatcagc gcgttggca actgaagca gattacgct tttataccg gaaaatcct gctgacgtg aactggcac gaaaaactc tgtcataag ttgtctcag gaacgccac atgggccaa cgtctcaat attattatc gtggctgaa ggcgctatc gaccggaat ggagaccca atcactgca gaaatgatc aaaaaagtc gtcgttgat aatctccaa caggacacc agaatcact gttttgggg cacgtgcaa cgtgggggc aacccgtct gcttttgat aggattctg ggctgtcgt atgggggct gaggccgtg atggcgcta atggaggct gatgagaat acagagccg tgtgtgatt tccttagac ggcaatcag gcgataagg ttgccgtta atggattgc gtgaagcgg acaaaagcc gtctcacag gcaatggct gataagaaa tttgatttg gcggtggaa ttacgagga aagtcattt atgcgcaat ttggagact tataagttg ttgactcga ctcaagcct ccaaaggat gcatttgat gagcaagga catggaagg gaaggtttc acagtggcc gtaatgcat attggagct cctgcttgt ggcatgaat gccgctgta cgttctttt gtacgaaat tttatctac agaggagac acagttttg ggcattcag gaaggagtt gaaggtctt atcgctggc aatataaaa caaatggac tgggccgaa gtggctggt tgggttggc atcgggggt gcaaatctg ggcaccaaa cgcaccctt cctggcaat cgtatgggt ttagtagcc gcccgattg gccgaattt aaaattcaa ggtttactc attattggc ggttttgaa gcatttgca gccgctgta caatttcta gacagccga aaagatttc ccagctctc aatatacca atcgtggta atccccgga actatcagt aacaatgta ccagggacg gaattttca ctaggttgt gacacagcc ttgaacgaa atcacagaa atttgtgat agaatcaga cagtcggct caaggcacg aaaaggagg gtttttatt attgaaacc atgggcggg tattgcgga tatttggcc acacttgca ggtctcgct gggggtgcg gatgctgcg tacatttac gaggaaaaa ttcacaatt aaagacttg caacaagat gtctaccac atggctacg aaaatgtcc gaaggggta caacgtggt ttaattttg cgtaatgaa aaggcgtca gataactac aataccgac tttatctac agattgtat tgtgaggaa ggaaaaggc ttgtttagt gccaggatg aacgtttta ggtcacatg caacaaggg ggttccccc acacctttt gaccgcaac gttggtacc aaaatggcg tccaaagcg gtcgaatgg atgatcaac caattaaaa agcaatatg agttcagac ggctcagtc agttgttct agtcaagac agcgtcgtt ctgttagga atcgttaaa agacaatac aagtttaca ccagtcgct gagttgaaa gagcaaacc aactttgac ttgcgtatg cctaaagag caatggtgg ttgaaactg cgccctcta ttgcgtatt ttggcaaaa catgattcg acatatgag gaagaaggg atgtatatg acgattgag gatcgcagt tttattaat taaaggcga ccgtagcca tttatagtg aatttgtat tttttagct tgttttatt tattttatt gtgtagttt attttatta aatgagata ttgtaaaca aattttggc catattgtg atgacaata tcgatattt gaagacggg tcaagtgtt acaaagtac gattcgagc tgggtatgt ggaatggac gtattatat ttcaataaa attttaata aagtatcaa attcttg
- >TC005855 RB
tgtcatatt ggactagag cagaggtca ccttgaatt aaaattgat tcctttttg aaactaagt ttgaattcg cgttctgaa atcccagtg aatgtcttg aagttaaag ccccctggt cagaataag agacaagaa gtcacgatg gaaggaaag gccgggcat aagtttgtg gggcgcgga gcccataag ggcaaaggc atcgctgtg tttaccagt ggaggagat tcacagggg atgaatgcg gctgtcagg gctgtggtc aggatgggg atttatttg gggtgcaaa gtctatttt atcaaggaa ggctaccaa ggcatggtt gatggcggc aagtacatc gttgaggcc aattggtcc tccgtaagc tctattatt cacaaagga ggcactgtg attggaagc gctcgttgc caagatttc cgtgaacgc gctggacgt ctcaaagct gcaaaaaac ctcgtagaa aacggcatt accaacctt gtagtaatt ggaggtgat ggtagttta acaggtgct gacctcttc cgtcaagaa tggagctct ctattggag gaattggcc accacaggt caaatcacg aaagaacag caacagaaa taccgcacg ttgcaaatt gccggtctt gtgggttcg attgataac gatttttgc ggtactgat atgaccatt gggacggac agtgccctt cacagaatt atcgaagct tgcgatgct atagtcagt acggcttat tcgcatcaa aggactttt attatggag gttatgggc cggcattgc gggtatttg gctcttgtg gccgcactc acttccgag gccgattat gtgttcata ccagaatca ccaccacca gacgattgg caaaaaagg ctatgcgat aaattagaa cagagcacc aaggacagg aacgaagga aaactcttc aaatcctgc cttggtaaa ccaccgatg ataatgatg aatgatttt aactggctt ctttcttct ttcttcttc caaggggcc ccaccccgc cccttggcg tggaacccg accttaagg caggattgt tatttagca ttgatcagc gcgttggca actgaagca gattacgct tttataccg gaaaatcct gctgacgtg aactggcac gaaaaactc tgtcataag ttgtctcag gaacgccac atgggccaa cgtctcaat attattatc gtggctgaa ggcgctatc gaccggaat ggagaccca atcactgca gaaatgatc aaaaaagtc gtcgttgat aatctccaa caggacacc agaatcact gttttgggg cacgtgcaa cgtgggggc aacccgtct gcttttgat aggattctg ggctgtcgt atgggggct gaggccgtg atggcgcta atggaggct gatgagaat acagagccg tgtgtgatt tccttagac ggcaatcag gcgataagg ttgccgtta atggattgc gtgaagcgg acaaaagcc gtctcacag gcaatggct gataagaaa tttgatttg gcggtggaa ttacgagga aagtcattt atgcgcaat ttggagact tataagttg ttgactcga ctcaagcct ccaaaggat gcatttgat gagcaagga catggaagg gaaggtttc acagtggcc gtaatgcat attggagct cctgcttgt ggcatgaat gccgctgta cgttctttt gtacgaaat tttatctac agaggagac acagttttg ggcattcag gaaggagtt gaaggtctt atcgctggc aatataaaa caaatggac tgggccgaa gtggctggt tgggttggc atcgggggt gcaaatctg ggcaccaaa cgcaccctt cctggcaat cgtatgggt ttagtagcc gcccgattg gccgaattt aaaattcaa ggtttactc attattggc ggttttgaa gcatttgca gccgctgta caatttcta gacagccga aaagatttc ccagctctc aatatacca atcgtggta atccccgga actatcagt aacaatgta ccagggacg gaattttca ctaggttgt gacacagcc ttgaacgaa atcacagaa atttgtgat agaatcaga cagtcggct caaggcacg aaaaggagg gtttttatt attgaaacc atgggcggg tattgcgga tatttggcc acacttgca ggtctcgct gggggtgcg gatgctgcg tacatttac gaggaaaaa ttcacaatt aaagacttg caacaagat gtctaccac atggctacg aaaatgtcc gaaggggta caacgtggt ttaattttg cgtaatgaa aaggcgtca gataactac aataccgac tttatctac agattgtat tgtgaggaa ggaaaaggc ttgtttagt gccaggatg aacgtttta ggtcacatg caacaaggg ggttccccc acacctttt gaccgcaac gttggtacc aaaatggcg tccaaagcg gtcgaatgg atgatcaac caattaaaa agcaatatg agttcagac ggctcagtc agttgttct agtcaagac agcgtcgtt ctgttagga atcgttaaa agacaatac aagtttaca ccagtcgct gagttgaaa gagcaaacc aactttgac ttgcgtatg cctaaagag caatggtgg ttgaaactg cgccctcta ttgcgtatt ttggcaaaa catgattcg acatatgag gaagaaggg atgtatatg acgattgag gatcgcagt tttattaat taaaggcga ccgtagcca tttatagtg aatttgtat tttttagct tgttttatt tattttatt gtgtagttt attttatta aatgagata ttgtaaaca aattttggc catattgtg atgacaata tcgatattt gaagacggg tcaagtgtt acaaagtac gattcgagc tgggtatgt ggaatggac gtattatat ttcaataaa attttaata aagtatcaa attcttg
