| General information | |||
| TC034166
Former TC numbers If one gene is splitted or genes are merged, the original TC numbers are listed. One listed former TC number indicates the previous gene has be splitted, serveral former TC number represents a merged gene. |
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: 12899927..12932652 | |
| Sequence information | |||
| Transcripts / Proteins (1) | Get mRNA sequence
Get CDS sequence Get protein sequence |
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| 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 |
|
TC034166 | |
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_09401 | 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 |
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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
Lethality 11 days after pupal injection: 50.0% (includes death as pupa, adult)
adult lethality - Number of animals showing the phenotype: 5 (9 dpi)Days post injection
Analysis of embryonic musculature and early development
number of eggs/embryos/larvae on slide: > 50
embryo/egg with muscle fluorescence/pattern, but not hatched (pot. late embryonic lethality) -
Penetrance (percentage of animals/eggs on slide): 50% - 80%
larval musculature pattern irregular - Penetrance (percentage of animals/eggs on slide): 30% - 50%
affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): more than 80%
larval musculature pattern irregular - Penetrance (percentage of animals/eggs on slide): 30% - 50%
affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): more than 80%
Analysis of larval stage 1 cuticle
number of eggs/embryos/larvae on slide: > 50
mesothoracic tracheal opening not present - Penetrance (percentage of animals/eggs on slide): more than 80%
abdomen tracheal opening size decreased - Penetrance (percentage of animals/eggs on slide): more than 80%
affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): more than 80%
mesothoracic tracheal opening not present - Penetrance (percentage of animals/eggs on slide): more than 80%
abdomen tracheal opening size decreased - Penetrance (percentage of animals/eggs on slide): more than 80%
affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): more than 80%
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)
- >TC034166 RA
MTHPNFQTN GASLEDCHT NFFALTELS GIKWRKLVY CELGGGGAA SGHCSGGGG GGEPLEDPV LRSYARCLA ADILCVWRR VSTPRSTDM FEPGPPPPP PLSLSTSKE LWIFWYGEE PDLGVLIAP ELNTPDSEQ GSWESGLSY ECRSLLFKA LHNLIERCL LSRDFIRLG KWFVQPYEG PKKNDFTMT HLSFSLAFF VHGESSVCA SVDVRQHPP VRRLTRLHL QQAQSSTSG LKVILAPYG LAGTLTGQS FKSSDQTSR LLEDWSHFY PLDKSSSLT DSSVVEVIV GGVKMKYPS CYVLVTDLD DNVNSAMNC TGVNPPNGL VKNGTSSPI LTQTPPPSP IQINSRVPQ PGPYSVSVE HLLSMSRDE TAATVLPER AWQECIHGG PPGCTPQPT SNTPHEFTG QWDFVDPIK RANCTCSKC RKYVGSMTS NKWSTGATT KTTNSPRPS KAVRGRVPF HRRSPVPAD SDRCSGSYT PQGGPPSYP RTQESVAIP SVGSPPSVA PSPLPNPHS QPASVPPGD QTMPMLSPH PPTLLPSPV EKTHTPDHP PKSVESTAN SPSCPTDAK NEQVTNVTS VPVLKRPVL SVKEYESSL GDEEQTLEM LYDYSTLEA WLNHPVKRF KPDGKENKR NRLGKNDIY SMYNHNGVT VNNNDVTNN FNTKNQFIK QEIKQEMTE SESTTPLGP MQGHGVKRP GDPYEFDEE GAGSACIDG FKRGQTVVK EEPKDSKKI STGNLFTSE GLQPSYKDL EQIFDNNSD DTSSDEAIQ VQTPPGSTT LNHHEDNKR LSGHGGCPT MGCLRPEEL SKMFPTPPS LEHNPIASP CGQMDLALP DYTDLGIVR TKQEIYPNM GSPPEEHID DWTYVFKPP VLCKMVGSS KYAPLTNLP SQSLPVVTL PSYCVYKPS WIQHAEKPL QQLPPPSTT QPSIPNNLH QNSFPPSPL HGGFRQPPP PYELPSPAT STASSYLNK NLNSVEAVP TPSVQRTPE ANSLALNIL LTDTASNIF RDHNFDSCT LCVCNAGGR VVGNIRGAD AGVYLISSS CDKTQSSVQ QPSSPFPGA CLGQPPPYG MLSPPHHHQ QQLLVDEDG IRCSCGFSA VVNRRLAHG SGLFYEDEM EITGIAEDP GEKKKPSLL SFLIASNSI KLDPSIDRD QVDVLPHGL LELIREQCV VIPNTANAL CRAAKYVCT KQTFNNIHV LEYSDSNEV TTIALEQSK SSYDLAVCK VEDKAKPAV GYAVHRWPF LRANGPQCN QDIVRVMRS LQPLLQEAI QKKCQTRLW EAPSAVKGP LTWRQFHRL AGRGTDDRC EPQPIPSVV VGHEKDWLS VSPYAIQHW ESLLLEPYS FARDVAYIV VAPDNDAIL PRVRTFFKE LSTAYEMCK LGRHSPITK VLRDGILRV GKTAKAKLG NEPVEEWFT LLGEGEITD MLKLYAQVC KHHLAPHLQ QVPMDKTLL DPPQESGDK PAPSPMPPP STPDGGQAA EKAPSTPKS DQDGDGMKE STPQANTNA ENSYDEDER EVPSVVVYL VEPFSLGSD QPELQRLAC LALLRCFQS VLAAVPENI RNNISVQII SLESIVELG RARDRTRLS DHMRALSLN IFSQCRRLL VHTNNVKSL TGFGTAAMA DLFLKSKDD KNRAAYKLY TPPYILAPM HEKIEAVES FGKGSGDQA SVLYLSYCL SEDQSWLLA VATDERGEI FETVTINID IPNRSRRKK ASARRIGLE KLMNFILGV MSQSVRPWR LVVGRLGRI GHGELKGWS WLLSRKNLL NASKNLKEI CNQCSLMYP TSVPCILSA CLVSLEPDS VLRLMPDQF TPDERFSQV SVNSQLSTP QDVSCTHIL VFPTSATTQ SSQTTFQEQ HITPELGDD ELFSAFTAE DMPEGIDSM NDFSDIFNW TDTGPGGGQ SPTGSPRRE ESGPGSPNC GVNNGRPGS PFHCGATNS KGNEQVEEV GTLLQQPLA LGYYVSTAP TGRMPKWFW ASCPHLEGI CPAFLKNAL HLHSPSIQQ SSDDLFQQA AVTSHPLDS QYTTDVLRY VLEGYNALS WLALDSNTK DRLSCLPVH MQVLLQLYH TTAALL
- >TC034166 RA
atgacgcac ccaaatttt caaaccaac ggggccagt ttagaggat tgccacact aacttcttc gctctgacg gaactttcg ggcatcaaa tggcgtaag ctggtgtat tgcgagttg gggggtggc ggtgccgcc tccggccat tgcagcggc ggcggagga ggcggcgaa ccccttgaa gatccggtg ttacgcagc tacgcccgt tgtctcgcc gccgatata ctttgcgtg tggcggcgg gtttcaacg ccacgttcg accgatatg ttcgaaccg ggaccacca ccgccgcca ccacttagt ctttcgaca tcgaaagaa ctatggatt ttctggtat ggagaggaa cccgatctg ggggttctt atcgcaccg gaacttaac actcctgat tcggagcag ggttcctgg gagagcggc ctgtcctac gagtgtcgg tctctcctc ttcaaagcc ttgcacaac ctgatcgag cgttgtctc ctctctcgc gattttatc aggttggga aaatggttc gttcagccg tacgaaggt ccaaagaag aacgacttc accatgact catttatcg ttctctttg gcgtttttc gtgcacggc gagagttcc gtatgtgca agcgtcgac gtacgccaa catccacca gttcgaaga ctcaccaga ttgcatttg caacaagct caatcctca acttctggt ctcaaagtg attttggct ccgtatgga cttgcgggg actttaacc ggtcaaagt ttcaagtct tcggaccaa acctcacgt ttgctggaa gattggagc cacttttat ccattggat aagagtagc agtttgacg gatagcagt gtggtggaa gtcattgtc ggaggtgtt aaaatgaag tatccgtcg tgctatgtt cttgtcacc gatctggac gataacgtc aattcggca atgaattgt acgggggtt aatccgcct aatgggctc gtcaagaac ggaacgagt tcaccgata ctgacccaa acaccaccc ccatctccg atccaaatc aacagtagg gtgccccaa ccagggccg tattcggtc tcggtggaa cacctctta tccatgtca cgcgacgaa actgccgcc actgttctg cccgaaagg gcatggcag gagtgcatc catgggggg cccccaggc tgcaccccc caaccaact tccaataca ccgcacgaa ttcaccggc caatgggat ttcgtcgat ccaatcaaa cgcgccaat tgtacctgt tctaagtgt agaaagtac gtgggcagc atgaccagc aataaatgg agcaccggg gccaccacc aagacaacc aactcgccc cggccttcg aaggcagta cggggacgt gtacctttc cacaggcgc tcccccgta cctgcagac tccgacaga tgctctggc tcctacacg cctcaaggg ggcccaccg tcgtaccct cgcacgcag gaatcggtg gccatccca tcggtgggg agcccccca tcggtggcc ccctcgcca ctacccaac ccccactcg caaccggct tcggtgccg cctggagac caaaccatg ccgatgttg tcgccccat ccgcccacg ttgcttccg tcaccagtt gaaaagaca cacaccccc gaccatccc cccaaaagc gttgaaagt acggcaaac agccccagt tgtccgact gatgcaaaa aacgaacaa gtgacgaac gtaacgagt gttcccgtg ttaaaaaga ccggttttg agtgtgaaa gaatacgaa agttcgtta ggggatgag gaacagact ttggaaatg ctttatgat tattcgacg ttagaagct tggttgaat catccggtt aagcggttt aagccggac ggtaaggaa aacaagagg aatagactc gggaaaaat gatatttac tcaatgtac aaccacaat ggtgtcact gtaaataac aacgacgta acgaacaat ttcaatacc aaaaatcaa tttattaag caggaaatt aagcaagaa atgactgaa agtgaatca acgacgccg ctcgggccg atgcaaggt catggggtt aagaggcct ggagatccg tacgagttc gatgaagag ggcgccggt tcagcctgc attgatggg ttcaaacgc gggcaaacg gtggtcaaa gaggaaccc aaagattcg aaaaaaatc agtactgga aatttgttt acaagcgaa ggcttgcaa ccgagctac aaagatctc gaacagatt tttgataac aattctgac gatacgagt agcgatgaa gctattcag gttcaaacg ccgccaggt tcgacgact ttgaaccat cacgaggat aataaaaga ctttcggga catggaggg tgtcccaca atggggtgt ttaaggccc gaagaattg agtaaaatg tttccaaca cctccaagt ctcgaacat aatccgatc gcgtcgcct tgcggacaa atggattta gcactgcca gattatacg gatttaggg atagtcagg accaaacag gagatttat cccaacatg gggagtccc ccggaggaa cacatagac gactggacg tacgttttc aagccgccc gtcttgtgt aaaatggtg ggttctagc aaatacgcc cctttgacg aacctgccc agtcaaagc cttcccgtg gtgaccctc ccatcgtac tgcgtctac aaaccatca tggatacaa cacgccgaa aaacccctc caacagctc ccaccaccg tccaccacc cagccttca atccccaac aacctccac caaaactcc ttccctccc agtccccta catggcggc ttccggcaa ccaccacct ccctacgag ctcccaagt ccggcaacg tcaaccgcc agttcctac cttaacaaa aacctgaat tcggtcgaa gctgttccc acaccgtca gtgcaaagg acacccgaa gcgaattcg ctcgctcta aacatcctc ctaaccgac accgcatcg aatattttc cgtgatcat aacttcgat agttgtacg ttgtgtgtg tgcaatgct ggcggccgt gtcgtcggt aacattcgt ggcgccgac gccggtgtt tatttgata agttcgtcc tgtgataaa acgcaaagt agtgttcaa caaccgagc agtccattc cccggggcc tgtttggga caaccgcct ccttacggc atgctctcg ccgccccac caccaccag cagcaactg ctcgtcgac gaggacggc attcggtgc agttgcggc ttctcggcc gtcgtcaac cgccggctg gcccacgga agtggactt ttctacgaa gacgagatg gaaatcacc ggcattgcc gaagacccc ggcgagaag aaaaaaccc tcgttgttg tcgttcctc atcgcgagc aattcgatt aaactcgac ccgagcatc gaccgggac caggtcgat gttttaccg cacggtctc ctggaattg atccgggaa cagtgcgtt gtgatcccg aatacggcg aacgccctc tgtcgggca gccaaatac gtctgcacc aagcaaact ttcaacaat attcacgtc ctcgaatac tccgatagt aacgaagtt acgacaata gccctggaa caaagtaaa agtagctac gacttggcc gtctgtaag gtcgaggat aaggcgaag ccggccgtt ggttacgcc gtccaccgg tggccgttc ctgcgggcc aacggcccg cagtgcaac caggacatc gtccgggtc atgcgctcc ctgcaaccg ctcctccag gaagccatc cagaagaaa tgccagacg cgcctctgg gaagccccc agtgcggtc aaaggcccg ctcacctgg cgccagttc caccggctt gccgggaga ggcaccgac gaccgctgc gagccccag cccataccg tccgtggtc gtcggccac gagaaggac tggctgtcg gtgtcgcct tatgccatc cagcactgg gaaagcttg ctgcttgag ccgtattcg ttcgcccgt gatgtggcg tatattgtg gtggcgccc gataacgat gcgatactt ccacgtgtt cggaccttt ttcaaggag ctgtccacg gcgtacgag atgtgcaag ttggggcgg catagtccc atcaccaaa gtgctgcgc gatgggatt ttaagggtg gggaagacg gccaaagct aaattaggg aatgaacct gtcgaggag tggtttacg ctgctgggg gaaggggaa attacggat atgttgaag ttgtacgcg caggtgtgt aagcatcat ctagcgcca catttgcaa caagtgccg atggataag acgttgctt gatccgcca caagagagt ggggataag ccggcaccg agtcccatg ccaccgccg agcacgccc gatggggga caggcggcg gagaaggcg cccagtacg ccgaagagt gaccaagac ggagacggc atgaaagaa tccacacct caggcaaac acaaacgcc gaaaattcg tacgatgag gacgaacgc gaggttccc tccgttgta gtttacctc gttgagcct ttctcacta ggttccgac caaccggaa ctccaaaga ctagcctgt ctggcactc cttcgttgt tttcagtcg gttcttgcc gccgtaccc gaaaacatc cgaaacaac atcagtgtc cagatcatt tctttggaa agtatcgtc gaattgggc cgtgccaga gatcggacg cgtctcagc gaccacatg cgcgccctc tccttgaac attttcagc caatgtcgg cgcctttta gtccacacg aacaacgtc aagtcgttg acaggcttt ggaacggct gctatggcc gatctgttt ttaaagagc aaagatgac aaaaatcgc gctgcctac aaactgtac actccgccg tacattctc gccccaatg cacgaaaaa atcgaagcg gtggagtcg ttcggcaaa ggcagcgga gaccaagcc tccgtcctc tatctcagt tactgtctc tcggaagat caaagctgg ctattggcc gtcgccacc gacgaacgt ggcgaaatt ttcgaaact gtcacaatc aacatagac ataccgaat cgtagcagg cggaaaaaa gcgtcagca cgtcgtatt ggccttgag aaattgatg aactttatc ttgggtgtg atgtcacaa agtgtgcgt ccatggcga ctggttgtt ggtcgcttg ggacggatc ggtcatggg gaattgaag ggttggagt tggttgttg agtcgaaaa aatctgcta aatgcgtcg aaaaatctc aaagagatt tgtaatcag tgctcgttg atgtatccg acttcggtt ccgtgtatt ttgagtgct tgtttagtg agtcttgag ccggattcg gtgttgaga cttatgcca gatcagttc acaccagat gaaaggttt agtcaagtc agtgttaat tcgcaattg tcgacaccg caagatgtg agctgtacg catattttg gtctttccg accagtgcc accacgcag tcgtcgcaa accacgttt caggaacag cacatcacg cctgagctc ggcgacgac gagctcttc agcgccttc accgccgaa gacatgccc gaaggcatc gacagcatg aacgacttc agcgacatc ttcaactgg acggacacg ggtcccgga ggcggccaa agtcccacg ggaagtcca agacgtgaa gaatcaggt cctggaagt cccaactgt ggggtgaac aacggccga ccaggcagt ccgttccac tgcggtgcc acgaactcc aaaggaaac gaacaagtc gaagaagtc ggcacgtta ttacagcaa cctctggcc ttaggctac tacgtctcg acggcaccc accggccgc atgccgaag tggttctgg gcgtcgtgc ccacacctc gaagggatt tgtccggcg tttctgaaa aacgccttg catttgcac agtccgagc atccagcag agttccgac gacttgttc cagcaggcg gccgtcacg tcgcatccg ctggactcg cagtacacc accgatgtt ttaagatac gtacttgaa ggctacaac gctctgtct tggctagcg ctggattcg aacacgaag gaccgattg tcttgtttg cctgtgcac atgcaggtc ttactgcag ttatatcac actacagca gcgcttttg taa
- >TC034166 RA
tgtctcggc tcttgttga attagtgtg gcttgttgt ttcctatcc gccattttg cattgcgtt cgaaaggtg aaaatcgag gtaaatggt ggcttaatt ggggcccca acgcgccag cgagtgcag tggaaagtt caagtgtcg tgggtgcta aatcgccga gaatggagc aggcatgac gcacccaaa ttttcaaac caacggggc cagtttaga ggattgcca cactaactt cttcgctct gacggaact ttcgggcat caaatggcg taagctggt gtattgcga gttgggggg tggcggtgc cgcctccgg ccattgcag cggcggcgg aggaggcgg cgaacccct tgaagatcc ggtgttacg cagctacgc ccgttgtct cgccgccga tatactttg cgtgtggcg gcgggtttc aacgccacg ttcgaccga tatgttcga accgggacc accaccgcc gccaccact tagtctttc gacatcgaa agaactatg gattttctg gtatggaga ggaacccga tctgggggt tcttatcgc accggaact taacactcc tgattcgga gcagggttc ctgggagag cggcctgtc ctacgagtg tcggtctct cctcttcaa agccttgca caacctgat cgagcgttg tctcctctc tcgcgattt tatcaggtt gggaaaatg gttcgttca gccgtacga aggtccaaa gaagaacga cttcaccat gactcattt atcgttctc tttggcgtt tttcgtgca cggcgagag ttccgtatg tgcaagcgt cgacgtacg ccaacatcc accagttcg aagactcac cagattgca tttgcaaca agctcaatc ctcaacttc tggtctcaa agtgatttt ggctccgta tggacttgc ggggacttt aaccggtca aagtttcaa gtcttcgga ccaaacctc acgtttgct ggaagattg gagccactt ttatccatt ggataagag tagcagttt gacggatag cagtgtggt ggaagtcat tgtcggagg tgttaaaat gaagtatcc gtcgtgcta tgttcttgt caccgatct ggacgataa cgtcaattc ggcaatgaa ttgtacggg ggttaatcc gcctaatgg gctcgtcaa gaacggaac gagttcacc gatactgac ccaaacacc acccccatc tccgatcca aatcaacag tagggtgcc ccaaccagg gccgtattc ggtctcggt ggaacacct cttatccat gtcacgcga cgaaactgc cgccactgt tctgcccga aagggcatg gcaggagtg catccatgg ggggccccc aggctgcac cccccaacc aacttccaa tacaccgca cgaattcac cggccaatg ggatttcgt cgatccaat caaacgcgc caattgtac ctgttctaa gtgtagaaa gtacgtggg cagcatgac cagcaataa atggagcac cggggccac caccaagac aaccaactc gccccggcc ttcgaaggc agtacgggg acgtgtacc tttccacag gcgctcccc cgtacctgc agactccga cagatgctc tggctccta cacgcctca agggggccc accgtcgta ccctcgcac gcaggaatc ggtggccat cccatcggt ggggagccc cccatcggt ggccccctc gccactacc caaccccca ctcgcaacc ggcttcggt gccgcctgg agaccaaac catgccgat gttgtcgcc ccatccgcc cacgttgct tccgtcacc agttgaaaa gacacacac ccccgacca tccccccaa aagcgttga aagtacggc aaacagccc cagttgtcc gactgatgc aaaaaacga acaagtgac gaacgtaac gagtgttcc cgtgttaaa aagaccggt tttgagtgt gaaagaata cgaaagttc gttagggga tgaggaaca gactttgga aatgcttta tgattattc gacgttaga agcttggtt gaatcatcc ggttaagcg gtttaagcc ggacggtaa ggaaaacaa gaggaatag actcgggaa aaatgatat ttactcaat gtacaacca caatggtgt cactgtaaa taacaacga cgtaacgaa caatttcaa taccaaaaa tcaatttat taagcagga aattaagca agaaatgac tgaaagtga atcaacgac gccgctcgg gccgatgca aggtcatgg ggttaagag gcctggaga tccgtacga gttcgatga agagggcgc cggttcagc ctgcattga tgggttcaa acgcgggca aacggtggt caaagagga acccaaaga ttcgaaaaa aatcagtac tggaaattt gtttacaag cgaaggctt gcaaccgag ctacaaaga tctcgaaca gatttttga taacaattc tgacgatac gagtagcga tgaagctat tcaggttca aacgccgcc aggttcgac gactttgaa ccatcacga ggataataa aagactttc gggacatgg agggtgtcc cacaatggg gtgtttaag gcccgaaga attgagtaa aatgtttcc aacacctcc aagtctcga acataatcc gatcgcgtc gccttgcgg acaaatgga tttagcact gccagatta tacggattt agggatagt caggaccaa acaggagat ttatcccaa catggggag tcccccgga ggaacacat agacgactg gacgtacgt tttcaagcc gcccgtctt gtgtaaaat ggtgggttc tagcaaata cgccccttt gacgaacct gcccagtca aagccttcc cgtggtgac cctcccatc gtactgcgt ctacaaacc atcatggat acaacacgc cgaaaaacc cctccaaca gctcccacc accgtccac cacccagcc ttcaatccc caacaacct ccaccaaaa ctccttccc tcccagtcc cctacatgg cggcttccg gcaaccacc acctcccta cgagctccc aagtccggc aacgtcaac cgccagttc ctaccttaa caaaaacct gaattcggt cgaagctgt tcccacacc gtcagtgca aaggacacc cgaagcgaa ttcgctcgc tctaaacat cctcctaac cgacaccgc atcgaatat tttccgtga tcataactt cgatagttg tacgttgtg tgtgtgcaa tgctggcgg ccgtgtcgt cggtaacat tcgtggcgc cgacgccgg tgtttattt gataagttc gtcctgtga taaaacgca aagtagtgt tcaacaacc gagcagtcc attccccgg ggcctgttt gggacaacc gcctcctta cggcatgct ctcgccgcc ccaccacca ccagcagca actgctcgt cgacgagga cggcattcg gtgcagttg cggcttctc ggccgtcgt caaccgccg gctggccca cggaagtgg acttttcta cgaagacga gatggaaat caccggcat tgccgaaga ccccggcga gaagaaaaa accctcgtt gttgtcgtt cctcatcgc gagcaattc gattaaact cgacccgag catcgaccg ggaccaggt cgatgtttt accgcacgg tctcctgga attgatccg ggaacagtg cgttgtgat cccgaatac ggcgaacgc cctctgtcg ggcagccaa atacgtctg caccaagca aactttcaa caatattca cgtcctcga atactccga tagtaacga agttacgac aatagccct ggaacaaag taaaagtag ctacgactt ggccgtctg taaggtcga ggataaggc gaagccggc cgttggtta cgccgtcca ccggtggcc gttcctgcg ggccaacgg cccgcagtg caaccagga catcgtccg ggtcatgcg ctccctgca accgctcct ccaggaagc catccagaa gaaatgcca gacgcgcct ctgggaagc ccccagtgc ggtcaaagg cccgctcac ctggcgcca gttccaccg gcttgccgg gagaggcac cgacgaccg ctgcgagcc ccagcccat accgtccgt ggtcgtcgg ccacgagaa ggactggct gtcggtgtc gccttatgc catccagca ctgggaaag cttgctgct tgagccgta ttcgttcgc ccgtgatgt ggcgtatat tgtggtggc gcccgataa cgatgcgat acttccacg tgttcggac ctttttcaa ggagctgtc cacggcgta cgagatgtg caagttggg gcggcatag tcccatcac caaagtgct gcgcgatgg gattttaag ggtggggaa gacggccaa agctaaatt agggaatga acctgtcga ggagtggtt tacgctgct gggggaagg ggaaattac ggatatgtt gaagttgta cgcgcaggt gtgtaagca tcatctagc gccacattt gcaacaagt gccgatgga taagacgtt gcttgatcc gccacaaga gagtgggga taagccggc accgagtcc catgccacc gccgagcac gcccgatgg gggacaggc ggcggagaa ggcgcccag tacgccgaa gagtgacca agacggaga cggcatgaa agaatccac acctcaggc aaacacaaa cgccgaaaa ttcgtacga tgaggacga acgcgaggt tccctccgt tgtagttta cctcgttga gcctttctc actaggttc cgaccaacc ggaactcca aagactagc ctgtctggc actccttcg ttgttttca gtcggttct tgccgccgt acccgaaaa catccgaaa caacatcag tgtccagat catttcttt ggaaagtat cgtcgaatt gggccgtgc cagagatcg gacgcgtct cagcgacca catgcgcgc cctctcctt gaacatttt cagccaatg tcggcgcct tttagtcca cacgaacaa cgtcaagtc gttgacagg ctttggaac ggctgctat ggccgatct gtttttaaa gagcaaaga tgacaaaaa tcgcgctgc ctacaaact gtacactcc gccgtacat tctcgcccc aatgcacga aaaaatcga agcggtgga gtcgttcgg caaaggcag cggagacca agcctccgt cctctatct cagttactg tctctcgga agatcaaag ctggctatt ggccgtcgc caccgacga acgtggcga aattttcga aactgtcac aatcaacat agacatacc gaatcgtag caggcggaa aaaagcgtc agcacgtcg tattggcct tgagaaatt gatgaactt tatcttggg tgtgatgtc acaaagtgt gcgtccatg gcgactggt tgttggtcg cttgggacg gatcggtca tggggaatt gaagggttg gagttggtt gttgagtcg aaaaaatct gctaaatgc gtcgaaaaa tctcaaaga gatttgtaa tcagtgctc gttgatgta tccgacttc ggttccgtg tattttgag tgcttgttt agtgagtct tgagccgga ttcggtgtt gagacttat gccagatca gttcacacc agatgaaag gtttagtca agtcagtgt taattcgca attgtcgac accgcaaga tgtgagctg tacgcatat tttggtctt tccgaccag tgccaccac gcagtcgtc gcaaaccac gtttcagga acagcacat cacgcctga gctcggcga cgacgagct cttcagcgc cttcaccgc cgaagacat gcccgaagg catcgacag catgaacga cttcagcga catcttcaa ctggacgga cacgggtcc cggaggcgg ccaaagtcc cacgggaag tccaagacg tgaagaatc aggtcctgg aagtcccaa ctgtggggt gaacaacgg ccgaccagg cagtccgtt ccactgcgg tgccacgaa ctccaaagg aaacgaaca agtcgaaga agtcggcac gttattaca gcaacctct ggccttagg ctactacgt ctcgacggc acccaccgg ccgcatgcc gaagtggtt ctgggcgtc gtgcccaca cctcgaagg gatttgtcc ggcgtttct gaaaaacgc cttgcattt gcacagtcc gagcatcca gcagagttc cgacgactt gttccagca ggcggccgt cacgtcgca tccgctgga ctcgcagta caccaccga tgttttaag atacgtact tgaaggcta caacgctct gtcttggct agcgctgga ttcgaacac gaaggaccg attgtcttg tttgcctgt gcacatgca ggtcttact gcagttata tcacactac agcagcgct tttgtaaac cactgaact agttggggc tttaattta tttcgattg ggcgagttg ttgactctg aatatcggc ttttgttgt taagtttta ctagtggtc gtgacgcgg caaagaacg aattgaata gaatactca tcgtgatgt aatagatat tttgttttg ttgtgtaac aatgccaat cccttttaa ttaagtgta caaacaagg aaattgtga tatttagag gttaattta agcgtaaat ccgaacgcc gaaagtttc attaactgc cagttactt cagtgtttt gcaaagtgt gttattgtt attttattt gttgttact ttgcggttt aaattttga atattgtaa agttgtaaa ttgtgtaca agaaataat ttatttttc tgttgttgt atataaaca tgtacaaaa gtgaaaccg agatactta gactgaact aggcaaatt actatcaaa attttatgt agaccactc atcgacatt tgttggtaa ataatcaaa ccaagtttg ccaaagaca gatggagat tggactttg ggtacaatc gtgtctatg tacataaac caacgtaag acattttag gacaatttg tacataagg aaatactta atcaaaagt ttctgtcaa tttgctacg ttttacaat agtcctagc aaatgttat aaccgatgt acgtatatt gaatgtaat aaatatgta ttaaatact ggggccaat ggtttaaac gtatcttat aaaacttgc ctcagaggt ttatcactc gggtgtttc agtggcatg ttgacggcc tgtgcatga tttcgttac ctatttagt aataatgga aagtcttac caacgtact taacaaaga tctcatcaa ccgtatgtt ggtaacgat gtttgtttc cctctttca ccatactgt gattttggt tgctctgtg gagtcggta atctatgca aaacaacta actcttaac agtatttta tatcttgta acgtctggc gattcgctc cttacttac ctgctatca ttatttata tacttattt aatttctac ataatatat ttttattat ggacggtat aggactgtt attgttcaa atagagaga cgttcaaca tttgtactg tacatacat agctactgt tgtataaga atatgtcaa ttttaggga attgggatt tatagttaa ataaatttg taaatattt tattgacat tagttgaaa tagtaaata cacataaaa tgttaat
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