| General information | |||
| TC034589 | 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_007418.3: 6626560..6647684 | |
| 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 |
|
TC034589 | |
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_00581 | 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
Lethality 11 days after pupal injection: 40.0% (includes death as pupa, adult)
eclosion not fulfilled (with pupal and adult features) - Number of animals showing the phenotype: 4 (3 dpi)Days post injection
Analysis of larval stage 1 cuticle
number of eggs/embryos/larvae on slide: < 20
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: 0.0% (includes death as larva, prepupa, pupa)
Lethalities 22 days after larval injection: 30.0% (includes death as larva, prepupa, pupa, adult)
Days post injection
eclosion delayed (still pupa) - Number of animals showing the phenotype: 1 (16 dpi)
Days post injection
adult lethality - Number of animals showing the phenotype: 3 (19 dpi)
Days post injection
- >TC034589 RA
MSEGQDSRA ERIARYKEQ RRKELAAQF NKIHIDSPT SQRRNSKDS NSSSSEGPR TTRTSRLRA AISSQENCS SPQKTKLGR EKTPPVTPT SNETGVSGS RRIERDKSS KRKSNLNRS LNSEEVPNL EGDDVKNRR RRRRFFPQE VLDQTADSS NDDKVKSDT SHTASFLSQ SSTSPNTAS PRLTPRKSE LSIHMENAR KGIAPSYSE MGRTKPAVK RPMVSSSPK DKFDNSMSD KEISRRASQ TKLNGENKT TSSRSLSDN DSSKNYKSI SKKMEELTA LTKETLAQV ERLASKNEE QPKQIKRPL SSSPKRIDK VSRNLATDR LHQPFSILK KKIAEEPVI VESVPNTAP VSILKRKVS QDDHKTEGA SSTHTPPVT FSPSVIEPA TTNRKQGIL KKRRSLDES QVMRHRSCS PDVANKAPD SRSILKNQR RSSMEELTR TQSPEQHLH GILKRKTSK TEDDDHSLN SPQGILKRR SGASSAGST SSTPHVSIT TAVILAAAG GAEMVLEGE SVKPILKKK SFSEESSYN DNSYCETPK PILKKKSST DTDECEERP KKPILKISR NSLERDSEC IKHGVPSES ECDVKPILK QNGKEEGAR PRLSFCGDT NDDSVIQFR SSRRSHTIC ADFNVKSDI KHKEEDRQL KKPRPLSVS ELVMNFENS TSTGAIPKR TSLKRTSDR YRTQPVTSN ELEASLNLV RSPQVEDTS KLPKSSPLL SPVANQSRS SFLDFTSSL ESDSNLNSF LSSLHNTSP DSGVCGKMS SDSAFQSLG DGLELDQDE DKPLEVASK PGKLETNLE LQMKAIAEE AQRKKLAKG GAGVRSYRN RKSYPSDAP SSSRFSTQP ITYDEVREA ARVKENSDG SEDKEEADP SNLSLHERI KFFSVKLKE NNSQKRDVP KRRPNRFQT QPVTSDEVE NARCISPTA FALEDGRSG DHTTSERRG ILKSSGSFR RGDSHPRSA LKKRDSFDG TSDYENRCS ILKSDVADV TYDEGISCN DSASDSEAA SVPCGVKTP EIKVEHSTA KRQTKVLKS SSETSDGES SGGREIRSI FKNENKLKV KQKLESVLS KSKSQNTFR DENESSNKK GSAIRRSQT QSTMPRAIE EIQAKLQQS GESGWKKRV QQNNNSSND LKLLNMNRY NDELSQRSI LEIKKDELD AASKQWKNR VEKSDADNF SVAGKMEKN REESNLQIN IAVDCDKSK KTPQAKRFK GKEGSEAFS TPVSPDKNR HVELSRSFS VPDPDQDNF GSESFLRTP GQKVIVSRP DDETYKSFY KSIEGKEIT EQIEVSQED FDLITRQSL LVQKKNVQV RRRRGASKN PIKALASRT DITNEYTEI ITGVAEREK KRQNIQKLA KSSSFATAA LAGLASKED FKSVALKKS RTPLTSVPY KDLMLLHVK GRQFVQTRL VEPVASSIN EGDNYILVT PSTLYNYTG KYSNVIEQS RAADIINHI QRSGDLGCQ EAEVVNINS ENCALPSKK IQEFWQKLK SEAGQTVSG GHHEEDGIY ESNILATNM IYNWRVTNW CLLTNFGVK CQNRDLEPP TNGKNAPPD KKKVAFKLA KEMWDLGYD YSECSVCPL NVASVLGNR PPVDVSRKG TSRPDWALF AKLTQHRET ILFKEKFLD WPDFSRVIK VKSDQTDKT VDASYNIKP CDVGQMLNR ESETPDLVI KNTHLGRGD QWFDEETKR LYDFETLKI SAWRVSENH PEQLSDETI GHFSNSHSY IIKWDYQTT VKGRDLSGK PSRYQKAGR DRCVYFCWQ GSNASINEK GAAALLTVE LDDEEAPQV RVVQGAEPA VFLKLFNGS MVVHDGKEE NRHKAKLFM VRGELEDEV CLMEVPCEM KELRSRTSF ILVLCELAK VVVWNGAKA SKQKRKVAK IAAENIVEK KADEFCWSN NNDCQVVVV EEGEENEEF LEGVEGGRG DYNALVGSD LDYDYTMRL FHLSALSGT FQAAEILCP HRSEFSSPY PFLQTELYS ASQPALFMI DNKHELWLW QGWWPEKER DEMELSDQT GSGAVRWQA ERKAAMQTA VSYWRESHN DDEPVPAYL VWAGLEPLE FKNLFPCWA DREDVAELN LKEGKRANA RLSVEDELA LLLRTTYPL AELLQRPLP EGVDPTHLE VYLSAEDFQ ELLSMTKEE FQKLPVWKQ TAMKKEKGL F
- >TC034589 RA
atgtctgag ggtcaggac agcagggct gaacgcatc gcccgatac aaggaacaa cggagaaaa gagctcgcc gcccaattt aataagata catatcgac tctccgact agtcaaaga cgcaacagc aaagacagc aactccagc agctcggag gggccgcgg accacccgg acctcgcgc ttgagggct gcaatctcg tcgcaagaa aattgttct agtccccag aagactaaa ctcggacgt gagaagact cctccagtc acaccaacc agtaatgaa acaggtgtc tctggaagt cgacgtatc gaacgtgac aaaagcagc aaacgaaaa tctaatctc aatcgctct ttaaactct gaagaggtc cccaatttg gaaggtgac gatgtaaag aaccgccga aggcgtcgc cggtttttc ccacaagaa gtattagac cagactgca gacagctcg aacgatgat aaagttaaa agtgataca agtcacact gcctctttc ttatcgcaa tcttctaca tcacccaac actgcctca cctcggctg acgcctcgc aagtcagaa ctaagcata cacatggaa aatgcgcga aaaggcata gccccatca tattccgag atgggacga acaaaacca gcagtcaaa cgtccgatg gtctcatcc agcccgaaa gataaattt gacaactcc atgtcagac aaagaaatc agtagaaga gcaagccaa actaaacta aatggggaa aataaaacc acgtcttct agaagttta tccgataac gatagtagc aagaattac aaaagtatt agtaaaaaa atggaggaa ctaacggcc ttaactaaa gaaacgctg gcccaggtc gagcgcctg gccagtaaa aacgaagaa cagcctaaa caaataaag aggccattg agtagtagt cctaaacga atagacaaa gttagccga aacttagct acagaccgg ctccaccaa ccattttcc attttaaaa aagaaaata gccgaggag cccgtcatt gtcgaatcc gtgccaaat accgctcca gtatcgata ctaaaacgc aaagtgtct caagacgac cacaaaacc gaaggtgcg tcttcgact cacactccc ccggtcact ttttcgccg agtgtcatt gaaccggcg acgacaaac cgcaaacaa ggcattttg aaaaaacgg cgaagtctg gacgagtct caagtcatg cggcatagg tcttgtagc cccgatgtg gccaacaag gctccagac tcccgctcc atactcaaa aaccagcgg aggtcctct atggaggag ttgactcgg acccagtcg cctgaacag cacctgcac gggatactt aagcgcaaa acgtcgaag actgaagac gatgatcat tcgctcaac tccccgcag gggattctg aagaggagg tcgggggca agcagtgcc gggtccact agtagcact ccgcatgtt tccatcacc acggctgtg attctggct gcggctgga ggcgcagag atggtgctt gaaggcgag agtgtgaag cccatcctc aagaagaag agtttttcg gaggagagt tcctacaat gataattct tattgcgag acgcctaag ccgattttg aagaagaag tcgagcacg gacactgat gagtgtgag gagaggccg aagaagccc attttgaag atatcccgc aattcttta gagcgtgac agtgagtgc atcaaacat ggtgttcct agcgaaagt gagtgcgac gttaaacca atactcaag caaaacggt aaagaggag ggggcgagg cctagattg tctttttgt ggcgataca aacgatgat tcagtcatt caatttcga agctcaaga aggtcgcat actatatgc gccgacttt aatgtcaag tctgatatt aagcacaag gaagaggac aggcagttg aagaagccg aggccgttg tccgtttcc gaacttgtg atgaatttt gaaaacagt acttccacg ggggcgatc ccgaaacgg acgagcttg aagagaacc agtgatagg tacaggacc cagccggtg acttcgaac gaactcgaa gcgagtttg aacttggtg aggagtccg caagtagaa gacacttca aaacttcca aaatcaagc ccgctgttg tctccagtc gccaatcaa agcaggagt tcgttcctg gattttacg tcttctttg gagagtgat agcaatctg aatagtttc ctgtcttcg cttcataat actagtccg gacagtgga gtttgtggt aagatgtct tccgattcc gcttttcaa tcgctcgga gacgggctt gagctagat caagacgag gataaacct ctggaagtt gcctcgaag cccggcaaa cttgaaact aatttggaa cttcaaatg aaagcaata gccgaagaa gcgcagcga aagaagtta gccaaaggc ggtgcaggg gttcgttcg tatcgcaac cgaaaatcc tacccatca gatgcgcct tcatctagc agattttcc actcagccc atcacttac gacgaagtt cgtgaagca gcaagagtc aaggagaat tccgacggt tctgaagac aaagaagaa gccgatcct tctaatcta agcttacac gaaaggatc aagttcttt agcgtcaag ctgaaagag aacaattcc caaaaacga gacgttcct aaacggcgc cctaatcgt tttcaaacc caacctgtt acttcagac gaggtcgag aatgcacgc tgtatttcc cccaccgca tttgcttta gaagatggg cgaagtggt gaccacaca acgagcgag agacgaggc attttgaag tcgtctggc agttttcgc cgcggcgat tcgcatccg cgatccgcg ctcaaaaaa cgcgattcg ttcgatggt acttccgac tacgaaaat cggtgctca attttgaaa tcggatgtt gcggatgtg acctacgac gaggggatt tcttgtaac gattctgcg tcggattcg gaagccgcg agtgttcct tgcggtgtt aaaaccccc gaaattaaa gtcgaacat agcacggca aagcggcaa actaaagtt ttgaaatcc agcagcgag acgtcagat ggcgaatcg tcgggggga cgcgaaatc aggagcatc ttcaagaat gaaaacaag ctgaaggtt aagcaaaaa ttagaaagt gttctctcc aaaagcaaa agccagaac accttcagg gatgaaaac gaatcatcc aacaaaaaa ggaagtgca atccgaaga tctcagact cagtcaacg atgcctcga gctattgaa gaaatccag gctaagctg cagcaaagc ggcgaatcc ggatggaaa aagagagtg caacagaac aataactcc agcaatgac ttaaaactg ctgaacatg aatcggtac aatgatgaa ctgtcgcaa cgatcaatt ttggagatc aagaaagat gaactggac gcagcatcg aaacagtgg aaaaaccgt gtcgaaaaa tcggacgcg gataatttc tctgtggcc ggaaaaatg gaaaaaaat cgagaagaa tcgaatctt caaatcaat atagctgta gactgtgat aaaagtaag aagacgccg caagccaag aggttcaaa ggcaaggaa gggtcggaa gctttctcc actccagtt tccccagac aagaaccgt cacgtggag ttatcaaga agtttttcc gtgccagac ccagaccag gataatttt ggttcggag tcgttttta agaacaccg ggtcaaaaa gtgatagtt tcaagaccg gacgatgag acgtacaaa agtttttac aagagcatt gaaggcaaa gaaataaca gaacaaata gaagtgtct caggaagat ttcgaccta atcacccga cagtctcta ctagttcaa aagaaaaat gtacaagtc cggagaaga cgtggtgct tccaaaaac ccgatcaag gctttggcc tcgcgcaca gacatcacc aacgaatac actgaaata atcacaggc gtagccgag agagaaaag aaaagacag aacatacaa aaacttgct aaaagttca agttttgcg acggccgct ttggccggg ttagcaagc aaagaagat tttaaatcc gtggctttg aaaaaatcg cggactcct ctcacatct gttccgtac aaagacctc atgttgtta cacgttaaa ggccgccag tttgtccaa acacgattg gtcgagcct gtggcttcg agtatcaac gaaggagat aattacatc ttggtcact ccttccact ttgtataat tacaccggg aagtattca aatgtgatt gaacagtca agggctgcc gatattatc aaccacatc caaagaagt ggcgatctg ggttgtcag gaagccgaa gttgttaac attaactct gaaaattgt gcgttacca agtaaaaag atacaagaa ttttggcaa aaattaaag agcgaagcg ggccaaact gttagtggc ggccaccat gaagaagat ggtatttac gagtccaac atactcgcc accaacatg atttataac tggagagta acgaactgg tgccttttg acgaatttt ggggtaaaa tgccaaaat cgagatctt gaaccaccg acgaacggc aaaaatgcc cctcctgat aaaaagaaa gtggccttc aagctggcc aaagaaatg tgggacctt ggctacgat tattccgaa tgtagcgtg tgtcccttg aatgtggct tcggtgctt ggcaatcgg cccccagtg gacgtctca cgcaaaggc acgtccaga cctgactgg gccctcttt gccaaactc acccaacac cgggaaaca attttattc aaggagaaa ttcctcgat tggcccgat ttctcccga gttatcaaa gtcaaaagc gaccaaacc gacaaaacc gtcgacgct tcctacaac atcaagccc tgtgatgtc ggccaaatg ctcaaccgt gagagcgaa acccccgac ctggtgatc aaaaacacg cacctgggg cgtggcgac cagtggttt gacgaagaa acgaaaaga ctgtacgac tttgaaacg ttgaaaata tcggcttgg agggtgtcc gagaaccac cccgagcag ctaagcgac gaaaccatc ggtcacttt tccaacagc cacagttac atcattaag tgggattac cagaccact gtgaaaggc cgcgatttg agcgggaag ccgtccagg taccagaaa gcgggtcgc gaccgttgc gtttatttt tgctggcaa ggatcgaac gcttcgatc aacgaaaaa ggggccgcc gctcttttg accgttgag ctcgacgac gaggaggcg ccccaggtg cgcgtggtg caaggggcg gagcccgcc gtgttcttg aaactgttt aacggctcc atggtggtg cacgacggc aaagaggaa aatcggcac aaggccaag ctgtttatg gtgcgaggt gagctggag gatgaagtt tgcctgatg gaggtcccg tgcgagatg aaggagttg cggagcaga acttcgttt atactcgtt ttgtgtgag ttggcgaaa gttgtggtg tggaatggg gccaaggcg agtaaacaa aagcggaag gtggcgaaa attgccgct gagaatatt gtggagaag aaggccgat gagttctgt tggagtaat aacaatgac tgtcaggtg gttgttgtg gaggaaggg gaggagaat gaggagttt ttggagggt gttgaggga ggtcgtggt gattataac gctttggtg ggcagtgat ttggattat gactacacc atgaggctt tttcacctt agtgctctt tcggggacg tttcaagct gccgaaatt ttgtgcccg cataggagt gaattttct agtccttat ccctttctg caaactgaa ctgtactca gctagtcaa cctgcgttg ttcatgatc gacaacaag cacgaattg tggctgtgg cagggctgg tggcccgag aaggaaaga gacgagatg gaactaagc gaccagacg ggctctgga gccgttcgg tggcaagcg gaaagaaaa gccgccatg caaacagct gtcagctac tggcgagag agccacaac gacgacgaa ccggtgcct gcttactta gtttgggca ggacttgag ccgctggaa tttaagaac ttgttcccg tgctgggcg gacagggaa gacgtggct gagttgaat ttgaaagag gggaaaagg gcgaatgcg aggctgtcg gtggaggac gagctggcg ctccttctg cggacgact taccccttg gccgagttg ctgcagcgg cctttgccg gaaggcgtc gatccgacc catctggag gtgtatttg tcggcggag gatttccag gagttgttg tcgatgact aaggaagag ttccagaaa ttgccggtg tggaaacag accgctatg aagaaggag aagggcctt ttttga
- >TC034589 RA
tctgtggtg tattaatag tttaaactg tgtggagta ttctgtgtt tatttcact gatgtactg tttggtgca gtgccgttg ttttaatta aaattttca tgtgcgctc aactactcg cgattattt tttattcat tttcatttg gagaagtca taatgtaaa catccgcca cgacgctca cgttctcgc ttacctcga tggctacga cctcattcc tctggaaat ttatcagac ccgacaatt atgtctgag ggtcaggac agcagggct gaacgcatc gcccgatac aaggaacaa cggagaaaa gagctcgcc gcccaattt aataagata catatcgac tctccgact agtcaaaga cgcaacagc aaagacagc aactccagc agctcggag gggccgcgg accacccgg acctcgcgc ttgagggct gcaatctcg tcgcaagaa aattgttct agtccccag aagactaaa ctcggacgt gagaagact cctccagtc acaccaacc agtaatgaa acaggtgtc tctggaagt cgacgtatc gaacgtgac aaaagcagc aaacgaaaa tctaatctc aatcgctct ttaaactct gaagaggtc cccaatttg gaaggtgac gatgtaaag aaccgccga aggcgtcgc cggtttttc ccacaagaa gtattagac cagactgca gacagctcg aacgatgat aaagttaaa agtgataca agtcacact gcctctttc ttatcgcaa tcttctaca tcacccaac actgcctca cctcggctg acgcctcgc aagtcagaa ctaagcata cacatggaa aatgcgcga aaaggcata gccccatca tattccgag atgggacga acaaaacca gcagtcaaa cgtccgatg gtctcatcc agcccgaaa gataaattt gacaactcc atgtcagac aaagaaatc agtagaaga gcaagccaa actaaacta aatggggaa aataaaacc acgtcttct agaagttta tccgataac gatagtagc aagaattac aaaagtatt agtaaaaaa atggaggaa ctaacggcc ttaactaaa gaaacgctg gcccaggtc gagcgcctg gccagtaaa aacgaagaa cagcctaaa caaataaag aggccattg agtagtagt cctaaacga atagacaaa gttagccga aacttagct acagaccgg ctccaccaa ccattttcc attttaaaa aagaaaata gccgaggag cccgtcatt gtcgaatcc gtgccaaat accgctcca gtatcgata ctaaaacgc aaagtgtct caagacgac cacaaaacc gaaggtgcg tcttcgact cacactccc ccggtcact ttttcgccg agtgtcatt gaaccggcg acgacaaac cgcaaacaa ggcattttg aaaaaacgg cgaagtctg gacgagtct caagtcatg cggcatagg tcttgtagc cccgatgtg gccaacaag gctccagac tcccgctcc atactcaaa aaccagcgg aggtcctct atggaggag ttgactcgg acccagtcg cctgaacag cacctgcac gggatactt aagcgcaaa acgtcgaag actgaagac gatgatcat tcgctcaac tccccgcag gggattctg aagaggagg tcgggggca agcagtgcc gggtccact agtagcact ccgcatgtt tccatcacc acggctgtg attctggct gcggctgga ggcgcagag atggtgctt gaaggcgag agtgtgaag cccatcctc aagaagaag agtttttcg gaggagagt tcctacaat gataattct tattgcgag acgcctaag ccgattttg aagaagaag tcgagcacg gacactgat gagtgtgag gagaggccg aagaagccc attttgaag atatcccgc aattcttta gagcgtgac agtgagtgc atcaaacat ggtgttcct agcgaaagt gagtgcgac gttaaacca atactcaag caaaacggt aaagaggag ggggcgagg cctagattg tctttttgt ggcgataca aacgatgat tcagtcatt caatttcga agctcaaga aggtcgcat actatatgc gccgacttt aatgtcaag tctgatatt aagcacaag gaagaggac aggcagttg aagaagccg aggccgttg tccgtttcc gaacttgtg atgaatttt gaaaacagt acttccacg ggggcgatc ccgaaacgg acgagcttg aagagaacc agtgatagg tacaggacc cagccggtg acttcgaac gaactcgaa gcgagtttg aacttggtg aggagtccg caagtagaa gacacttca aaacttcca aaatcaagc ccgctgttg tctccagtc gccaatcaa agcaggagt tcgttcctg gattttacg tcttctttg gagagtgat agcaatctg aatagtttc ctgtcttcg cttcataat actagtccg gacagtgga gtttgtggt aagatgtct tccgattcc gcttttcaa tcgctcgga gacgggctt gagctagat caagacgag gataaacct ctggaagtt gcctcgaag cccggcaaa cttgaaact aatttggaa cttcaaatg aaagcaata gccgaagaa gcgcagcga aagaagtta gccaaaggc ggtgcaggg gttcgttcg tatcgcaac cgaaaatcc tacccatca gatgcgcct tcatctagc agattttcc actcagccc atcacttac gacgaagtt cgtgaagca gcaagagtc aaggagaat tccgacggt tctgaagac aaagaagaa gccgatcct tctaatcta agcttacac gaaaggatc aagttcttt agcgtcaag ctgaaagag aacaattcc caaaaacga gacgttcct aaacggcgc cctaatcgt tttcaaacc caacctgtt acttcagac gaggtcgag aatgcacgc tgtatttcc cccaccgca tttgcttta gaagatggg cgaagtggt gaccacaca acgagcgag agacgaggc attttgaag tcgtctggc agttttcgc cgcggcgat tcgcatccg cgatccgcg ctcaaaaaa cgcgattcg ttcgatggt acttccgac tacgaaaat cggtgctca attttgaaa tcggatgtt gcggatgtg acctacgac gaggggatt tcttgtaac gattctgcg tcggattcg gaagccgcg agtgttcct tgcggtgtt aaaaccccc gaaattaaa gtcgaacat agcacggca aagcggcaa actaaagtt ttgaaatcc agcagcgag acgtcagat ggcgaatcg tcgggggga cgcgaaatc aggagcatc ttcaagaat gaaaacaag ctgaaggtt aagcaaaaa ttagaaagt gttctctcc aaaagcaaa agccagaac accttcagg gatgaaaac gaatcatcc aacaaaaaa ggaagtgca atccgaaga tctcagact cagtcaacg atgcctcga gctattgaa gaaatccag gctaagctg cagcaaagc ggcgaatcc ggatggaaa aagagagtg caacagaac aataactcc agcaatgac ttaaaactg ctgaacatg aatcggtac aatgatgaa ctgtcgcaa cgatcaatt ttggagatc aagaaagat gaactggac gcagcatcg aaacagtgg aaaaaccgt gtcgaaaaa tcggacgcg gataatttc tctgtggcc ggaaaaatg gaaaaaaat cgagaagaa tcgaatctt caaatcaat atagctgta gactgtgat aaaagtaag aagacgccg caagccaag aggttcaaa ggcaaggaa gggtcggaa gctttctcc actccagtt tccccagac aagaaccgt cacgtggag ttatcaaga agtttttcc gtgccagac ccagaccag gataatttt ggttcggag tcgttttta agaacaccg ggtcaaaaa gtgatagtt tcaagaccg gacgatgag acgtacaaa agtttttac aagagcatt gaaggcaaa gaaataaca gaacaaata gaagtgtct caggaagat ttcgaccta atcacccga cagtctcta ctagttcaa aagaaaaat gtacaagtc cggagaaga cgtggtgct tccaaaaac ccgatcaag gctttggcc tcgcgcaca gacatcacc aacgaatac actgaaata atcacaggc gtagccgag agagaaaag aaaagacag aacatacaa aaacttgct aaaagttca agttttgcg acggccgct ttggccggg ttagcaagc aaagaagat tttaaatcc gtggctttg aaaaaatcg cggactcct ctcacatct gttccgtac aaagacctc atgttgtta cacgttaaa ggccgccag tttgtccaa acacgattg gtcgagcct gtggcttcg agtatcaac gaaggagat aattacatc ttggtcact ccttccact ttgtataat tacaccggg aagtattca aatgtgatt gaacagtca agggctgcc gatattatc aaccacatc caaagaagt ggcgatctg ggttgtcag gaagccgaa gttgttaac attaactct gaaaattgt gcgttacca agtaaaaag atacaagaa ttttggcaa aaattaaag agcgaagcg ggccaaact gttagtggc ggccaccat gaagaagat ggtatttac gagtccaac atactcgcc accaacatg atttataac tggagagta acgaactgg tgccttttg acgaatttt ggggtaaaa tgccaaaat cgagatctt gaaccaccg acgaacggc aaaaatgcc cctcctgat aaaaagaaa gtggccttc aagctggcc aaagaaatg tgggacctt ggctacgat tattccgaa tgtagcgtg tgtcccttg aatgtggct tcggtgctt ggcaatcgg cccccagtg gacgtctca cgcaaaggc acgtccaga cctgactgg gccctcttt gccaaactc acccaacac cgggaaaca attttattc aaggagaaa ttcctcgat tggcccgat ttctcccga gttatcaaa gtcaaaagc gaccaaacc gacaaaacc gtcgacgct tcctacaac atcaagccc tgtgatgtc ggccaaatg ctcaaccgt gagagcgaa acccccgac ctggtgatc aaaaacacg cacctgggg cgtggcgac cagtggttt gacgaagaa acgaaaaga ctgtacgac tttgaaacg ttgaaaata tcggcttgg agggtgtcc gagaaccac cccgagcag ctaagcgac gaaaccatc ggtcacttt tccaacagc cacagttac atcattaag tgggattac cagaccact gtgaaaggc cgcgatttg agcgggaag ccgtccagg taccagaaa gcgggtcgc gaccgttgc gtttatttt tgctggcaa ggatcgaac gcttcgatc aacgaaaaa ggggccgcc gctcttttg accgttgag ctcgacgac gaggaggcg ccccaggtg cgcgtggtg caaggggcg gagcccgcc gtgttcttg aaactgttt aacggctcc atggtggtg cacgacggc aaagaggaa aatcggcac aaggccaag ctgtttatg gtgcgaggt gagctggag gatgaagtt tgcctgatg gaggtcccg tgcgagatg aaggagttg cggagcaga acttcgttt atactcgtt ttgtgtgag ttggcgaaa gttgtggtg tggaatggg gccaaggcg agtaaacaa aagcggaag gtggcgaaa attgccgct gagaatatt gtggagaag aaggccgat gagttctgt tggagtaat aacaatgac tgtcaggtg gttgttgtg gaggaaggg gaggagaat gaggagttt ttggagggt gttgaggga ggtcgtggt gattataac gctttggtg ggcagtgat ttggattat gactacacc atgaggctt tttcacctt agtgctctt tcggggacg tttcaagct gccgaaatt ttgtgcccg cataggagt gaattttct agtccttat ccctttctg caaactgaa ctgtactca gctagtcaa cctgcgttg ttcatgatc gacaacaag cacgaattg tggctgtgg cagggctgg tggcccgag aaggaaaga gacgagatg gaactaagc gaccagacg ggctctgga gccgttcgg tggcaagcg gaaagaaaa gccgccatg caaacagct gtcagctac tggcgagag agccacaac gacgacgaa ccggtgcct gcttactta gtttgggca ggacttgag ccgctggaa tttaagaac ttgttcccg tgctgggcg gacagggaa gacgtggct gagttgaat ttgaaagag gggaaaagg gcgaatgcg aggctgtcg gtggaggac gagctggcg ctccttctg cggacgact taccccttg gccgagttg ctgcagcgg cctttgccg gaaggcgtc gatccgacc catctggag gtgtatttg tcggcggag gatttccag gagttgttg tcgatgact aaggaagag ttccagaaa ttgccggtg tggaaacag accgctatg aagaaggag aagggcctt ttttgacct aaactggct tcgttgttg gacaaacta gtctatttg tttggtggg tgtttccgc actacatct atcaatttt atttgaaaa cttgatttt agtgtgttt gacatatca aaattattt tattcgtat ccagtgtga aataaaagc tgattgcct ttccaaatt atattttta taactggta gctttcata caaaattgt tttttattg ataactccg tgtaataaa actgccaat tgatatgat ttacctgta cttaagtga tttgttttg atgtttttc aatgacaat tttgagtca aatttatag tgtctgtga ttaggcttt tgtattcat aatgataac cgtaatata tgctgttat gtaaaatgt atatacatt gaagaggtg ccttgttat ctacggaat tgagagcac tcctccatt cgtgagttg ccaatattt tctagtctt tgtgatctt ttgtggcac tgcgttatt tttttttat ttcaatgtt ttttattca ttgtgtagt acctattta attaccctc tactaatgt tgcgttcac acaatgttt gtcaatatg agcgataat tgtacattc atttttaaa cgctgtgta cattataac tggcttttg tactaaaat taatatact gtggacgtt ctgtgtttt gtatataac ttttcaatt acttaataa atattttca gaaaaa
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