| General information | |||
| TC009268 | 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: 2158244..2184055 | |
| 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 |
|
TC009268 | |
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_01518 | 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 |
||
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.
Metamorphosis and survival
Fertility
vitellarium shortened -
Number of animals showing the phenotype: 4
mature egg not present - Number of animals showing the phenotype: 4
vitellogenic egg chamber number decreased - Number of animals showing the phenotype: 4
mature egg not present - Number of animals showing the phenotype: 4
vitellogenic egg chamber number decreased - Number of animals showing the phenotype: 4
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: 100.0% (includes death as larva, prepupa, pupa)
Lethalities 22 days after larval injection: 100.0% (includes death as larva, prepupa, pupa, adult)
Days post injection
- >TC009268 RA
MIGPHHVCV LYVATAGLQ GSLMGSDEQ EVVLLIYVI VDVVQNKIV GVQQYPIRP QTADINENI LSEQLTSES VLTDELIKS SGRQLEDAL NEFDAYCNS LHIDPHSAG FRLLTDGQL PLRQCLYPE ACRKEIDLP RYYSTFHDL RKEVARFTR KTEEVPQSV ADMLEYLQI KPEVENEFY KREAKDMVN IVQRLISEG YRFEALETI NLTLEPGIC SKDDEIDSN CVVRARGLP WQSSDQDIA KFFSGLNVA KGGVALCLS AQGRRNGEA LIKFESQEH RDMALKRHK HHIGPRYIE VYRASGEDF LSVAGGKSS EAQAFLSRG AAVIVRMRG LPYDCTAKQ VLDFFASGE NSCTVLDGT EGVLFVRKP DGRATGDAF VLFASESDA PKALSKHRE CIGSRYIEL FRSTTAEVQ QVLNRSLEA RTYEPEPLP PLLPHMPLL PQHYITSGT KKECIRLRG LPYEAQVEH ILEFLGEYA KNIIFQGVH MVYNAQGQP SGEAFIQMD SEQSACITA QQKHHRYMT FGKKQRYIE VFQCSGEDM NLVLTGGIP APVSPAKAT PALLSPGML PSIAPLPPT NIPPPPPPN TSVNQISPN ITNWDNQAL FAQQQAQII AQQNLIARQ NQAQAQNEM LLMNQIAQH NLAVLNQNV GPVSPNSPN SVLKNTPMM PHLIAQPPF VMLPPRMAP TLGLPRPQL AYPQHLLPG GAHGVFPVM PHVAVKRSY GDAFNEQAA PAKRAFHAQ GALPVYQHF YPNM
- >TC009268 RB
MIGPHHVCV LYVATAGLQ GSLMGSDEQ EVVLLIYVI VDVVQNKIV GVQQYPIRP QTADINENI LSEQLTSES VLTDELIKS SGRQLEDAL NEFDAYCNS LHIDPHSAG FRLLTDGQL PLRQCLYPE ACRKEIDLP RYYSTFHDL RKEVARFTR KTEEVPQSV ADMLEFRTR RTLQFCEYL RLTYKPDLQ IKPEVENEF YKREAKDMV NIVQRLISE GYRFEALET INLTLEPGI CSKDDEIDS NCVVRARGL PWQSSDQDI AKFFSGLNV AKGGVALCL SAQGRRNGE ALIKFESQE HRDMALKRH KHHIGPRYI EVYRASGED FLSVAGGKS SEAQAFLSR GAAVIVRMR GLPYDCTAK QVLDFFASG ENSCTVLDG TEGVLFVRK PDGRATGDA FVLFASESD APKALSKHR ECIGSRYIE LFRSTTAEV QQVLNRSLE ARTYEPEPL PPLLPHMPL LPQHYITSG TKKECIRLR GLPYEAQVE HILEFLGEY AKNIIFQGV HMVYNAQGQ PSGEAFIQM DSEQSACIT AQQKHHRYM TFGKKQRYI EVFQCSGED MNLVLTGGI PAPVSPAKA TPALLSPGM LPSIAPLPP TNIPPPPPP NTSVNQISP NITNWDNQA LFAQQQAQI IAQQNLIAR QNQAQAQNE MLLMNQIAQ HNLAVLNQN VGPVSPNSP NSVLKNTPM MPHLIAQPP FVMLPPRMA PTLGLPRPQ LAYPQHLLP GGAHGVFPV MPHVAVKRS YGDAFNEQA APAKRAFHA QGALPVYQH FYPNM
- >TC009268 RA
atgatcggg ccccaccat gtttgcgtc ctgtacgtg gccaccgcg gggctccag ggctccctc atgggctcc gacgagcag gaggtggtg ctcctcatc tatgtcatc gtcgacgtc gtccagaat aagattgtg ggcgtgcag cagtaccca atcaggccg caaaccgcc gacatcaac gaaaatatc ctatcggaa cagctcaca tctgagtcg gttttgacc gacgaactc ataaaatca tctggaagg caattggag gacgccctc aacgaattc gatgcctac tgcaatagt ttacacata gacccgcac agtgccggc ttcaggctc ctaacagac ggccaatta ccgttaagg cagtgtctg tatccggaa gcatgtagg aaggaaatt gatctgccg cgttattat agcacgttc catgacctc aggaaagaa gttgccagg tttacgaga aagacggaa gaagtaccc caaagtgtt gccgatatg ttagaatat ttacaaatt aaacccgaa gttgagaat gaattctac aagagggag gctaaggat atggtcaat attgtacaa aggctgatt tcggaagga tacaggttc gaggcgctc gaaacgata aacctgacc ctggagccc ggaatatgt tccaaagac gatgagatt gacagcaac tgcgtggtg agggcgaga ggacttccc tggcagagc agcgatcag gacattgcc aagtttttc agcggactc aatgtcgcc aagggcggc gtagctctg tgcttgtcg gcacaagga cgacggaac ggcgaagct ttaataaaa ttcgagagt caagagcac cgcgacatg gctttgaag aggcacaag caccacatc ggcccccga tacatcgaa gtgtatcgc gcttccggc gaggacttc ctctcggtg gctgggggc aagagttcc gaagcccag gccttcctg tcccggggg gctgccgtc atcgtgcgc atgcggggg ctgccctac gactgcacc gccaaacaa gtcttggat tttttcgcg agtggcgag aactcgtgc accgttttg gacggcact gaaggcgtc ctgttcgtg cggaagccc gacggccga gccactggc gacgcgttc gttctgttt gcgagcgaa tcggacgcg cctaaagct ctctcgaaa catagggaa tgtataggt tctaggtat atcgaattg ttcaggtcg acgacggcc gaggtgcaa caggtgctt aacaggtcg ctggaagcg cggacttac gagccggaa ccgttgccc ccgttatta ccgcacatg cctttattg cctcaacat tatattacg tctgggacc aaaaaggaa tgcattagg ttaagagga ctgccttac gaagctcaa gtcgaacac atcctagag tttctcgga gaatatgcc aaaaatatt atattccaa ggggtgcat atggtctac aatgcacag ggccagccc agcggcgag gccttcatc cagatggac tccgagcag tccgcctgc ataaccgcc cagcagaag caccaccgc tacatgaca ttcggcaaa aagcagcgc tacatcgaa gtattccaa tgctccggc gaggacatg aatctcgta ctgaccggc ggcattccc gcccccgtc tcgcccgca aaggccacc cccgcgcta ctcagtccc ggtatgtta ccttcaatt gcgcctctt ccacctact aatatcccc ccgccgccc ccgccaaac acctcggtc aaccaaata tccccaaac atcacaaac tgggacaac caggccctt ttcgcccaa cagcaggcc cagatcata gcccagcaa aacctgatc gcgcgccaa aaccaagcg caagcccag aacgaaatg ctcctaatg aaccagatt gcgcaacac aatctggcc gtactcaac caaaatgtg ggcccagtg tccccaaac agccccaat tcggtgctg aaaaacacg ccgatgatg ccgcatttg atcgcgcag cccccgttt gtcatgcta cccccgagg atggccccc actttgggg ctccccagg ccccagttg gcctacccc cagcatttg ctgcccggg ggcgcgcat ggggttttt ccggtcatg ccccacgtt gcggttaaa cgcagctat ggggacgcg ttcaatgag caagccgcc ccagccaaa agggccttc catgcacaa ggggccttg cccgtctac caacacttc taccctaac atgtag
- >TC009268 RB
atgatcggg ccccaccat gtttgcgtc ctgtacgtg gccaccgcg gggctccag ggctccctc atgggctcc gacgagcag gaggtggtg ctcctcatc tatgtcatc gtcgacgtc gtccagaat aagattgtg ggcgtgcag cagtaccca atcaggccg caaaccgcc gacatcaac gaaaatatc ctatcggaa cagctcaca tctgagtcg gttttgacc gacgaactc ataaaatca tctggaagg caattggag gacgccctc aacgaattc gatgcctac tgcaatagt ttacacata gacccgcac agtgccggc ttcaggctc ctaacagac ggccaatta ccgttaagg cagtgtctg tatccggaa gcatgtagg aaggaaatt gatctgccg cgttattat agcacgttc catgacctc aggaaagaa gttgccagg tttacgaga aagacggaa gaagtaccc caaagtgtt gccgatatg ttagaattc cgcactcgt cgtacgctt cagttttgc gaatattta cggctaacc tacaaacca gatttacaa attaaaccc gaagttgag aatgaattc tacaagagg gaggctaag gatatggtc aatattgta caaaggctg atttcggaa ggatacagg ttcgaggcg ctcgaaacg ataaacctg accctggag cccggaata tgttccaaa gacgatgag attgacagc aactgcgtg gtgagggcg agaggactt ccctggcag agcagcgat caggacatt gccaagttt ttcagcgga ctcaatgtc gccaagggc ggcgtagct ctgtgcttg tcggcacaa ggacgacgg aacggcgaa gctttaata aaattcgag agtcaagag caccgcgac atggctttg aagaggcac aagcaccac atcggcccc cgatacatc gaagtgtat cgcgcttcc ggcgaggac ttcctctcg gtggctggg ggcaagagt tccgaagcc caggccttc ctgtcccgg ggggctgcc gtcatcgtg cgcatgcgg gggctgccc tacgactgc accgccaaa caagtcttg gattttttc gcgagtggc gagaactcg tgcaccgtt ttggacggc actgaaggc gtcctgttc gtgcggaag cccgacggc cgagccact ggcgacgcg ttcgttctg tttgcgagc gaatcggac gcgcctaaa gctctctcg aaacatagg gaatgtata ggttctagg tatatcgaa ttgttcagg tcgacgacg gccgaggtg caacaggtg cttaacagg tcgctggaa gcgcggact tacgagccg gaaccgttg cccccgtta ttaccgcac atgccttta ttgcctcaa cattatatt acgtctggg accaaaaag gaatgcatt aggttaaga ggactgcct tacgaagct caagtcgaa cacatccta gagtttctc ggagaatat gccaaaaat attatattc caaggggtg catatggtc tacaatgca cagggccag cccagcggc gaggccttc atccagatg gactccgag cagtccgcc tgcataacc gcccagcag aagcaccac cgctacatg acattcggc aaaaagcag cgctacatc gaagtattc caatgctcc ggcgaggac atgaatctc gtactgacc ggcggcatt cccgccccc gtctcgccc gcaaaggcc acccccgcg ctactcagt cccggtatg ttaccttca attgcgcct cttccacct actaatatc cccccgccg cccccgcca aacacctcg gtcaaccaa atatcccca aacatcaca aactgggac aaccaggcc cttttcgcc caacagcag gcccagatc atagcccag caaaacctg atcgcgcgc caaaaccaa gcgcaagcc cagaacgaa atgctccta atgaaccag attgcgcaa cacaatctg gccgtactc aaccaaaat gtgggccca gtgtcccca aacagcccc aattcggtg ctgaaaaac acgccgatg atgccgcat ttgatcgcg cagcccccg tttgtcatg ctacccccg aggatggcc cccactttg gggctcccc aggccccag ttggcctac ccccagcat ttgctgccc gggggcgcg catggggtt tttccggtc atgccccac gttgcggtt aaacgcagc tatggggac gcgttcaat gagcaagcc gccccagcc aaaagggcc ttccatgca caaggggcc ttgcccgtc taccaacac ttctaccct aacatgtag
- >TC009268 RA
tagctaaaa aagtgctgc aatacaagt ggctatttt tgaaccagt ttagtgatt tccaatgat ctttcgcga aatacttaa ctaaacacc gacctgaaa gcctgaaaa agaaaaata gaaataggt attttaagg cttgccaac ctgaaaaca aaaaagaca accaattgc agccaccag tcattctga tgtgcactt tagaacatt ttgcggaaa aactgatgg gaatttatg agtggcata taaacgcag ctttgtgat gctctaggt gctcagtct ttatgacat ccataaact aggtgatgc cttgaaaag tcaacacca atgcaccaa taaatagac tgcggtcag tttggcacc ggagtggag atttcgatg ctcgatcac atcacatta aatgaattt ttgcttagt atttgtcga attaggtta tgttcgctt ccccattcg ccatatgct ttcgccagt cgacttgat aaaacgcta atcactcgt cttgtactc gcagcatca attcttgtt gcctgaaag tgttgctca tcacaatga ccattggca caagcaagc gaaaatgtg cagaacatg cggcaaccg accaaagtt atctaatta tataaccat agaatatgc ggctcgaat ccagttacc ttgcgttag actgtacat acccttctc cacgattgg ctcacggct caggtagaa ccagtctac gcggcccct tcatacgta cacgggaaa attgaggct acgagttac ctgcacgac aaaattact cgaaacttc caaagttcg gcgactgtt aaaccaaac ccgtcgtca attgcgtag aaggctgtg aacgcgttt tgtcattaa tttttcaaa caattgccg atcgtcgac aacctgttg atcgcagtg gccaggtgt gtagtgaca gtgactgtt ttagtgacg ccaactgcg aaatcgcca aaaatcgca ctgtgatcg aaatcccta ggttagagc gaatgtctg tgatagcat ctccggaga taagatagc cccaggtgc cccacgttc cccgaaatc caacaggtg tcacgtggc aacatgtcg ggctggtga tggtgtgct tcagagcca ggatgaggt gctgaggcc cctttggtg gacaaatga gcattaaag gcctcaaca tgatcgggc cccaccatg tttgcgtcc tgtacgtgg ccaccgcgg ggctccagg gctccctca tgggctccg acgagcagg aggtggtgc tcctcatct atgtcatcg tcgacgtcg tccagaata agattgtgg gcgtgcagc agtacccaa tcaggccgc aaaccgccg acatcaacg aaaatatcc tatcggaac agctcacat ctgagtcgg ttttgaccg acgaactca taaaatcat ctggaaggc aattggagg acgccctca acgaattcg atgcctact gcaatagtt tacacatag acccgcaca gtgccggct tcaggctcc taacagacg gccaattac cgttaaggc agtgtctgt atccggaag catgtagga aggaaattg atctgccgc gttattata gcacgttcc atgacctca ggaaagaag ttgccaggt ttacgagaa agacggaag aagtacccc aaagtgttg ccgatatgt tagaatatt tacaaatta aacccgaag ttgagaatg aattctaca agagggagg ctaaggata tggtcaata ttgtacaaa ggctgattt cggaaggat acaggttcg aggcgctcg aaacgataa acctgaccc tggagcccg gaatatgtt ccaaagacg atgagattg acagcaact gcgtggtga gggcgagag gacttccct ggcagagca gcgatcagg acattgcca agtttttca gcggactca atgtcgcca agggcggcg tagctctgt gcttgtcgg cacaaggac gacggaacg gcgaagctt taataaaat tcgagagtc aagagcacc gcgacatgg ctttgaaga ggcacaagc accacatcg gcccccgat acatcgaag tgtatcgcg cttccggcg aggacttcc tctcggtgg ctgggggca agagttccg aagcccagg ccttcctgt cccgggggg ctgccgtca tcgtgcgca tgcgggggc tgccctacg actgcaccg ccaaacaag tcttggatt ttttcgcga gtggcgaga actcgtgca ccgttttgg acggcactg aaggcgtcc tgttcgtgc ggaagcccg acggccgag ccactggcg acgcgttcg ttctgtttg cgagcgaat cggacgcgc ctaaagctc tctcgaaac atagggaat gtataggtt ctaggtata tcgaattgt tcaggtcga cgacggccg aggtgcaac aggtgctta acaggtcgc tggaagcgc ggacttacg agccggaac cgttgcccc cgttattac cgcacatgc ctttattgc ctcaacatt atattacgt ctgggacca aaaaggaat gcattaggt taagaggac tgccttacg aagctcaag tcgaacaca tcctagagt ttctcggag aatatgcca aaaatatta tattccaag gggtgcata tggtctaca atgcacagg gccagccca gcggcgagg ccttcatcc agatggact ccgagcagt ccgcctgca taaccgccc agcagaagc accaccgct acatgacat tcggcaaaa agcagcgct acatcgaag tattccaat gctccggcg aggacatga atctcgtac tgaccggcg gcattcccg cccccgtct cgcccgcaa aggccaccc ccgcgctac tcagtcccg gtatgttac cttcaattg cgcctcttc cacctacta atatccccc cgccgcccc cgccaaaca cctcggtca accaaatat ccccaaaca tcacaaact gggacaacc aggcccttt tcgcccaac agcaggccc agatcatag cccagcaaa acctgatcg cgcgccaaa accaagcgc aagcccaga acgaaatgc tcctaatga accagattg cgcaacaca atctggccg tactcaacc aaaatgtgg gcccagtgt ccccaaaca gccccaatt cggtgctga aaaacacgc cgatgatgc cgcatttga tcgcgcagc ccccgtttg tcatgctac ccccgagga tggccccca ctttggggc tccccaggc cccagttgg cctaccccc agcatttgc tgcccgggg gcgcgcatg gggtttttc cggtcatgc cccacgttg cggttaaac gcagctatg gggacgcgt tcaatgagc aagccgccc cagccaaaa gggccttcc atgcacaag gggccttgc ccgtctacc aacacttct accctaaca tgtagcaag tacgggcgg aaagttttc gcaacttaa agtgcctta aagtgacaa ataatttga aaaaaaaaa ggatatttt ttacggttt attatactt tttttttct aaaacacga ttttgaaaa tttttttta cctcgtcca gtggtactt tttgatttt cacattcga actactgta tatcagtga tatttattg gttatttta ttgttagtt taacagtta agaataata gataaggaa atgacctaa ttttgacga tttatagag ttagtacca aaaatcact ctcataaac tttttgtac atttgtata tattacgaa ttggtactc tgataaata aaaagtgcc tcaaaaaga catgtaatt gataattac tggctttat tccgaaatc gggttttgc cacaaagag taccatttt ctgtggtac tatttttcg aatttctga acattatcg ttaatgtag gaatcgtga ataattgat aagagaaaa acgaagctc acatttggt tgtaagttt tttattctt gcgtatttg tataattac ggattgaaa ctaaaatcg gataataaa ttatttgtt ttattttta ataaataaa gcgtttcca tacaa
- >TC009268 RB
tagctaaaa aagtgctgc aatacaagt ggctatttt tgaaccagt ttagtgatt tccaatgat ctttcgcga aatacttaa ctaaacacc gacctgaaa gcctgaaaa agaaaaata gaaataggt attttaagg cttgccaac ctgaaaaca aaaaagaca accaattgc agccaccag tcattctga tgtgcactt tagaacatt ttgcggaaa aactgatgg gaatttatg agtggcata taaacgcag ctttgtgat gctctaggt gctcagtct ttatgacat ccataaact aggtgatgc cttgaaaag tcaacacca atgcaccaa taaatagac tgcggtcag tttggcacc ggagtggag atttcgatg ctcgatcac atcacatta aatgaattt ttgcttagt atttgtcga attaggtta tgttcgctt ccccattcg ccatatgct ttcgccagt cgacttgat aaaacgcta atcactcgt cttgtactc gcagcatca attcttgtt gcctgaaag tgttgctca tcacaatga ccattggca caagcaagc gaaaatgtg cagaacatg cggcaaccg accaaagtt atctaatta tataaccat agaatatgc ggctcgaat ccagttacc ttgcgttag actgtacat acccttctc cacgattgg ctcacggct caggtagaa ccagtctac gcggcccct tcatacgta cacgggaaa attgaggct acgagttac ctgcacgac aaaattact cgaaacttc caaagttcg gcgactgtt aaaccaaac ccgtcgtca attgcgtag aaggctgtg aacgcgttt tgtcattaa tttttcaaa caattgccg atcgtcgac aacctgttg atcgcagtg gccaggtgt gtagtgaca gtgactgtt ttagtgacg ccaactgcg aaatcgcca aaaatcgca ctgtgatcg aaatcccta ggttagagc gaatgtctg tgatagcat ctccggaga taagatagc cccaggtgc cccacgttc cccgaaatc caacaggtg tcacgtggc aacatgtcg ggctggtga tggtgtgct tcagagcca ggatgaggt gctgaggcc cctttggtg gacaaatga gcattaaag gcctcaaca tgatcgggc cccaccatg tttgcgtcc tgtacgtgg ccaccgcgg ggctccagg gctccctca tgggctccg acgagcagg aggtggtgc tcctcatct atgtcatcg tcgacgtcg tccagaata agattgtgg gcgtgcagc agtacccaa tcaggccgc aaaccgccg acatcaacg aaaatatcc tatcggaac agctcacat ctgagtcgg ttttgaccg acgaactca taaaatcat ctggaaggc aattggagg acgccctca acgaattcg atgcctact gcaatagtt tacacatag acccgcaca gtgccggct tcaggctcc taacagacg gccaattac cgttaaggc agtgtctgt atccggaag catgtagga aggaaattg atctgccgc gttattata gcacgttcc atgacctca ggaaagaag ttgccaggt ttacgagaa agacggaag aagtacccc aaagtgttg ccgatatgt tagaattcc gcactcgtc gtacgcttc agttttgcg aatatttac ggctaacct acaaaccag atttacaaa ttaaacccg aagttgaga atgaattct acaagaggg aggctaagg atatggtca atattgtac aaaggctga tttcggaag gatacaggt tcgaggcgc tcgaaacga taaacctga ccctggagc ccggaatat gttccaaag acgatgaga ttgacagca actgcgtgg tgagggcga gaggacttc cctggcaga gcagcgatc aggacattg ccaagtttt tcagcggac tcaatgtcg ccaagggcg gcgtagctc tgtgcttgt cggcacaag gacgacgga acggcgaag ctttaataa aattcgaga gtcaagagc accgcgaca tggctttga agaggcaca agcaccaca tcggccccc gatacatcg aagtgtatc gcgcttccg gcgaggact tcctctcgg tggctgggg gcaagagtt ccgaagccc aggccttcc tgtcccggg gggctgccg tcatcgtgc gcatgcggg ggctgccct acgactgca ccgccaaac aagtcttgg attttttcg cgagtggcg agaactcgt gcaccgttt tggacggca ctgaaggcg tcctgttcg tgcggaagc ccgacggcc gagccactg gcgacgcgt tcgttctgt ttgcgagcg aatcggacg cgcctaaag ctctctcga aacataggg aatgtatag gttctaggt atatcgaat tgttcaggt cgacgacgg ccgaggtgc aacaggtgc ttaacaggt cgctggaag cgcggactt acgagccgg aaccgttgc ccccgttat taccgcaca tgcctttat tgcctcaac attatatta cgtctggga ccaaaaagg aatgcatta ggttaagag gactgcctt acgaagctc aagtcgaac acatcctag agtttctcg gagaatatg ccaaaaata ttatattcc aaggggtgc atatggtct acaatgcac agggccagc ccagcggcg aggccttca tccagatgg actccgagc agtccgcct gcataaccg cccagcaga agcaccacc gctacatga cattcggca aaaagcagc gctacatcg aagtattcc aatgctccg gcgaggaca tgaatctcg tactgaccg gcggcattc ccgcccccg tctcgcccg caaaggcca cccccgcgc tactcagtc ccggtatgt taccttcaa ttgcgcctc ttccaccta ctaatatcc ccccgccgc ccccgccaa acacctcgg tcaaccaaa tatccccaa acatcacaa actgggaca accaggccc ttttcgccc aacagcagg cccagatca tagcccagc aaaacctga tcgcgcgcc aaaaccaag cgcaagccc agaacgaaa tgctcctaa tgaaccaga ttgcgcaac acaatctgg ccgtactca accaaaatg tgggcccag tgtccccaa acagcccca attcggtgc tgaaaaaca cgccgatga tgccgcatt tgatcgcgc agcccccgt ttgtcatgc tacccccga ggatggccc ccactttgg ggctcccca ggccccagt tggcctacc cccagcatt tgctgcccg ggggcgcgc atggggttt ttccggtca tgccccacg ttgcggtta aacgcagct atggggacg cgttcaatg agcaagccg ccccagcca aaagggcct tccatgcac aaggggcct tgcccgtct accaacact tctacccta acatgtagc aagtacggg cggaaagtt ttcgcaact taaagtgcc ttaaagtga caaataatt tgaaaaaaa aaaggatat tttttacgg tttattata ctttttttt tctaaaaca cgattttga aaatttttt ttacctcgt ccagtggta ctttttgat tttcacatt cgaactact gtatatcag tgatattta ttggttatt ttattgtta gtttaacag ttaagaata atagataag gaaatgacc taattttga cgatttata gagttagta ccaaaaatc actctcata aactttttg tacatttgt atatattac gaattggta ctctgataa ataaaaagt gcctcaaaa agacatgta attgataat tactggctt tattccgaa atcgggttt tgccacaaa gagtaccat tttctgtgg tactatttt tcgaatttc tgaacatta tcgttaatg taggaatcg tgaataatt gataagaga aaaacgaag ctcacattt ggttgtaag ttttttatt cttgcgtat ttgtataat tacggattg aaactaaaa tcggataat aaattattt gttttattt ttaataaat aaagcgttt ccatacaa
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