| General information | |||
| TC007517 | 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_007419.2: 3440471..3451158 | |
| 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 | TC007517 | ||
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_04410 | 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.
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: 30.0% (includes death as larva, prepupa, pupa)
Lethalities 22 days after larval injection: 100.0% (includes death as larva, prepupa, pupa, adult)
Days post injection
pupal molt delayed (still larva) - Number of animals showing the phenotype: 7 (11 dpi)
Days post injection
larva lethality late (L7) - Number of animals showing the phenotype: 7 (16 dpi)
Days post injection
- >TC007517 RA
MTALDGQPQ PSGQPFRPV HTYVMNPKS ITMGQLYGE FDLQTHEWT DGILPCLVR IGVAAENKD KRWYVFDGP VDAVWIENM NTVLDDNKK LCLSSGEII KLRDTMTMM FEVADLAVA SPATVSRCG MVYLEPGVL GLAPFINCW LKRLPKLAA PFADKFKEL IYAYVVPGI ELVRNRLRE ILTSVDSAL LYKFLRLMD YWLLPLSGR DNKPPPGPQ FLALIPELL APWVIFSMI WSIGCTCDN RSRVRFDHW LRGKMLEEG DEPYFPQKG LVYDYRLHD GGFTDPTED GNPMPPHWY SWMDNVEEF KITVDMKYS DMEVPTMDT VRNAKMLEI VLNNYDNVL CVGPTGTGK SLTVVGKLS RSMHKKFIC DFMSFSART SANQTQDLL DSKLDRRRK GVFGPPVLK RQIFFIDDF NMPALEVYG AQPPIELIR QWMDFGGWY DRKNIGDFR TIIDTNFCV AMGPPEGGR NPVTARLLR HFHYLAFIE MEDDSKRKI FGTIFKFWM ERIPEEFTL GHFYEPILE ATLNVYSTI LKELLPTPA KTHYTFNLR DLSKVFQGL LMFNPEKMT DINLLVRLW YHECCRIFQ DRLVNDDDR NWFANLLKN KIESEFEVD PVSALGTQM LFFADFIDP TTDYREYEQ ITDSARLSN VLDYYLNEY NEQSTRPMK LVLFLDAIS HVCRISRII RQPLGNALL LGVGGSGRQ SLTRLATFM AEYVCFQIE IHKTYGMFE WREDIKGLM LKAGLHRRE TVFLFSDTQ IKSESFLED LNNVLNSGD VPNIYQPDE LDKIFQEMR GVVQEMGLV ATKSNLFAT YQKVVRANL HTVVTMSPI GDIFRARLR QFPALVNCC TIDWFSEWP DSALRSVAF QFLDDMAEL DVGQEVLHG IVMVCQFMH ASVVDASIL YKQELSRHN YVTPTSYLE LLSSYTDLM LKKRGALSE GITRLSTGL DKLKSTAEE VKILQASLE EMKPALEAA KQDAETMIA EIAKDQIIA EETKEIVEK EELEAAKKA LETQTIADD AQRDLDEAM PDLLAAEAS LKSLNKNDI SEVRSMKRP PAGVVYVIE SICIIKNIK PAKLPGRMP GEKVLDYWE PGRNMLADP GVFLTSLMN FDKESITEE MIDKLKNYV SNPQFQPAK VAKVSKACK SFCMWVHAM YKYYFVNKR VAPKKAALA QAKAELAKT EAALAAAKA KMQAVLDNL RLLQEKLGA KIAFKEEKE ASIQICLER MSRAVRLIN GLSDERVRW LNTISSIEA SVVNVTGDI LLSAGAVAY LTPFTDKYR RGLLSEWLK EIKEQKIPC SADPNPVSV LAEPVQIRA WQLDGLPRD YFSTENAVL VSNSKRWPL FIDPQGQAN KWVKNMERA QGLAVCKMA DKELLRTLE SAVRFGKPV LIEGVGIDL DPSLDPVLL RQKFKQAGT WMLKLGEVV VPYDDDFHL YMTTKLPNP HYTPEVSVK VLLVNFTLV PSGLQDQLL ALVVMQERP DLEDQRSQL IVGSTQMKQ ELKEIEDRI LYKLSSLEG SPLDDLDFI ITLEASKVK SDDIKNKVV AAEITQIDI DNTRALYIP VANRAQILF FCLSDLSNV DPMYQYSLE WFVGIFIAS MAETERTEN IDERVVIIN EYFTFSLYS NVCRSLFEK HKLHFAFLM CARIFMDMG KIDGHEWHH FLAGGTPLS EDPNPAPDW LSGKAWKEI LAMRVLPNY VRFVDTFAQ YKEDYRVIF ESGDPHRKP LPEEFEKSF SEFQKLLVL KCLRPDKVT NAMQDFISS KMGQRFIEP QTTDLSVVF KDSGPTTPL IFVLSTGTD PAADLYKFA DKMKFAKRM FSISLGQGQ GPIAEKMLH NGTELGSWI FFQNCHLAP SWMPRLERL VENISPEIV HRDFRIWLT STPSPHFPV SILQNGSKM TIEPPAGIK ANIMRAYTN QVSDLIDIL HGDGPKNFN FKWLLFSLC LFHGVLLER RKFGPLGFN IPYEFTDGD LKICISQLR MFLWEYDEI PFKVLTYTA GHINYGGRV TDDWDRRCL MNILADYYK PEVVNADYV FDAHGFYHQ MPAETPYYD YLDYIKHFP INDDPELFG MHPNADISF AQAQTYSCL STLLTLQPR QVGGAAASE EEVIAQTAA SILNHLPRQ FDLEAISEK YPVLYEESL NTVLIQEGI RFNRLLKAI ESSLKDLLK AIKGLVVMS EALDKMATS LFSNIVPAL WSSKAYPSL KPLGAWVAD LDARVKFLN TWVDQGIPS VFWISGFYF PQAFLTATL QNFARKYVV SIDTINFSF KVLEHAPAK RPDDGCCIW GLFVEGARW NSSVGTLDE SHPKELYTE MPVVWLLPE ENHVKPEGR YECPVYKTL TRAGVLSTT GHSTNYVLA IEVPSKKSE AHWIKRGVA LICALDY
- >TC007517 RA
atgacggcg ctcgatggg cagccccag cccagcggc caaccattc cgtcctgtg cacacctac gtcatgaac cccaaatcc atcacaatg ggccaatta tacggcgag tttgatttg cagactcac gaatggacg gacgggatt ctcccgtgt ttggtgcgc attggagtg gctgcggag aacaaagac aagcgttgg tacgttttc gacgggccc gtggacgct gtctggatc gaaaacatg aataccgtc cttgacgat aataaaaaa ttgtgcttg agcagcggc gaaataatc aaactgcga gatacaatg acgatgatg tttgaagtt gcagattta gctgtagct tctccggcg actgtctcc cgatgcggg atggtttac ctcgagccg ggagttttg ggattagcg ccgtttatc aactgttgg ttgaaacga ctcccaaag ctggcggcg cctttcgcc gacaaattc aaagagttg atttatgca tatgtggtg ccagggatt gaactagtc cgtaaccgg cttagggag atcttgact tctgttgat tcggccctt ttgtacaaa tttctacgt ttgatggat tactggctt ttgccgtta tcggggcgt gataataaa ccgccccca ggaccacag tttttagct ttgattccc gagcttttg gctccttgg gtcattttt tcgatgatt tggagcatc ggctgcact tgcgataac cggagtcgc gtacgtttc gaccattgg ctcagggga aaaatgcta gaggaagga gatgagccg tactttccc cagaaaggt ctggtgtac gattataga ttgcacgat ggtgggttt actgaccct acggaggat ggaaatccc atgccaccg cactggtac agctggatg gacaatgtg gaagaattt aaaatcaca gtggacatg aaatattcc gatatggaa gttccgact atggatact gtacgaaac gccaaaatg ttggaaata gttttgaac aactacgat aatgtgtta tgtgtcggc ccaacggga actggaaaa tctctcaca gttgtcgga aagttgagc cgaagtatg cacaagaag tttatttgc gattttatg agtttctca gcgaggact tcggctaat caaacccag gatttgctc gattcaaaa ctcgatcga cgaagaaaa ggcgtgttt ggacctccg gtcttaaaa cgccagatc tttttcatt gatgatttt aacatgccg gctttagaa gtgtacggt gcccaacca cccatcgaa ctaattcgt caatggatg gattttgga ggttggtat gaccggaaa aacatcggc gactttcgc acaattatt gatacgaat ttttgtgtc gcgatggga ccccctgaa gggggcaga aacccggtc acggctaga ttattgaga catttccat tatttggcc ttcatcgaa atggaggat gatagtaag aggaaaatt ttcggaaca atttttaaa ttttggatg gagcggata ccggaagag tttaccttg gggcatttt tacgaaccg attctggaa gcaacactt aatgtttat agcacgatt ttaaaagaa ctgcttcct acgccagca aaaacgcac tatactttt aatttgcgg gatttgagt aaggttttt caagggttg ctaatgttt aatcctgaa aaaatgacg gacataaat ttattggtg cggttgtgg taccacgag tgttgtcgc attttccaa gaccgatta gtgaacgac gacgacaga aactggttt gcaaatttg ctcaaaaat aaaatcgag agtgagttt gaagttgat cctgtttcg gccttaggg actcagatg ttgtttttt gccgatttt attgatcca acgactgat tatcgcgaa tatgaacaa attactgat tcagcacgg ttatcaaac gtccttgat tactatttg aacgagtat aacgagcag agcacgcgt ccgatgaag ctggtcctt ttcctggat gcaatttcg cacgtttgt cgcatttcg aggatcatt cggcagcca ctaggcaat gcacttctg ttgggtgta ggcggatcc ggacgacag agtctgacg agacttgcg acctttatg gcggagtac gtttgtttc cagattgaa attcacaaa acttacggc atgtttgaa tggcgcgag gacatcaaa ggactaatg cttaaagcc ggtcttcac agacgcgaa actgttttt cttttctca gacacacaa attaaatcc gagtcgttc ctagaagac ttaaataat gtactaaat tccggagac gtcccgaat atttatcaa ccggacgaa ttggataag attttccaa gaaatgaga ggagtcgtg caagaaatg ggactcgtg gcaacaaaa tctaattta tttgccact tatcagaaa gtagtgaga gcaaatttg cacacagtg gtcacaatg agtccgata ggtgatatt ttccgggca agactacga cagtttccg gctttagtc aattgttgc acgatcgac tggtttagc gaatggcca gattcagcc cttaggtcg gtagcgttc caatttttg gatgacatg gctgaatta gacgtagga caagaagtc ttgcatgga attgtcatg gtgtgtcaa ttcatgcat gcaagtgtt gtcgatgcg agcattctt tacaagcaa gagctttcg agacataat tatgtcacg ccaacgtct tatctcgaa cttttgtca agctatact gacctcatg ttgaaaaaa agaggagct ttatcggag ggcattacg agattgtca accggcttg gataagctc aaaagcact gccgaagaa gtaaagatc cttcaggcc agccttgaa gaaatgaaa ccggccttg gaagccgca aaacaagac gccgaaaca atgattgct gaaatcgcc aaagatcaa atcatcgca gaagagacg aaagaaatt gtcgaaaag gaagagctc gaggcggcc aaaaaagct ctcgaaacc caaacaatc gccgatgac gctcaacga gatttagat gaggcaatg ccggatctg ttagcggcc gaagctagt ttaaaatcg cttaataaa aacgatatt tccgaagtg cgatcgatg aagcgtccg ccggccggc gtagtatac gtcattgaa tcgatttgc atcattaaa aacatcaaa ccggcgaaa ttacccgga agaatgccc ggagagaaa gttttagac tactgggag ccgggtcgc aacatgctc gctgatccg ggcgtgttt ctaacgtcg ttaatgaac tttgacaaa gaatcaatc accgaggaa atgatcgac aagctgaaa aattacgta tcgaatcca caatttcag cctgcaaaa gtggccaag gtctctaaa gcgtgcaaa tcgttctgt atgtgggtt catgcaatg tataaatac tactttgta aataagcga gtggcgccg aagaaggcc gccttggca caggcgaaa gccgagttg gccaaaacc gaagccgct ttagccgca gccaaggca aaaatgcaa gcggttttg gataattta agattgttg caagaaaaa ttaggggcg aaaattgct ttcaaagag gagaaagaa gcgagtatt caaatttgt ttggaacgg atgagtcga gcagttcgc ctaatcaat ggattatcc gatgagaga gttcgctgg ttgaacacc atttcgagc attgaagca agtgttgtc aatgtcacg ggtgatata ctcctgagt gccggtgct gtggcttat ctcacccca tttacggat aaatacaga cgagggttg ttgtccgaa tggctcaaa gagataaag gagcaaaag ataccctgt tcggctgat cctaatccg gtttcggtt ttagccgaa ccggtccag atacgcgca tggcaattg gacggctta ccgagggat tatttctca acggagaac gctgttctc gtttcgaat tcgaaacgt tggcctttg tttatagat ccacaggga caggccaat aaatgggtt aagaatatg gaaagggct caaggattg gcggtgtgt aaaatggcc gataaggag ctgttgcgt actttagag tcggcggtt aggtttggt aaaccagtt ttgattgag ggagtcgga attgattta gatccgtct ctggatcca gttttactg cgccaaaaa tttaaacag gcaggaacg tggatgttg aagctgggt gaagtcgtc gtgccttac gatgacgat tttcattta tacatgaca acgaaatta ccgaatccg cattatacg cctgaagtt tcagttaag gttcttttg gtgaatttt actctggtc ccgagtggc ttgcaggac cagctgctg gctttagtt gtcatgcag gagcgtccc gatttggag gaccagaga agtcagttg attgtcgga agtacacag atgaaacag gaattgaag gaaattgaa gacaggatt ttgtacaag ttgagttca ttggaaggt tcacctttg gatgattta gattttata ataacactc gaagcatcg aaagttaag agcgacgat attaaaaat aaagtggtc gctgctgaa ataacacaa atcgacatc gacaataca cgtgccttg tacattccc gtagcaaat agggcacag atattgttt ttctgtttg tccgattta tcaaacgtc gaccctatg tatcaatac tcgttggag tggttcgtt ggtattttc atagcaagc atggccgag acggaacga accgaaaat attgatgaa agagttgtc ataatcaac gaatacttt acgttcagt ttgtatagt aatgtttgt cgcagtttg tttgaaaaa cacaaatta cattttgcg tttttgatg tgcgcccgg atatttatg gacatgggg aaaattgac gggcacgaa tggcatcac tttttggca ggaggcacc cctctctcg gaagatcca aatcccgcg cctgattgg ctctcgggc aaagcatgg aaggaaatc ctcgcaatg cgtgtcctt ccaaattac gttcgattc gtcgatact ttcgcacaa tacaaagaa gattatcgc gtcatattt gagtctggg gaccctcac agaaaaccg ttacccgaa gagttcgaa aaatcattc tccgagttt caaaagcta ttagttttg aagtgtttg cgtcccgac aaagtgaca aacgccatg caagatttc atttcgtcc aaaatgggt cagagattt atcgaacca cagacgacc gacttatcc gtcgtcttc aaagactcc gggcctaca acccctttg attttcgtc ctgtccact ggaacggat ccggccgca gatctgtac aagttcgca gacaagatg aaattcgcc aagagaatg ttttcgatc tcgctgggc caaggccaa ggacccatt gccgagaaa atgctacac aacgggact gaactcggc tcgtggatt ttcttccag aactgtcac ctggcgccc agttggatg ccgcggttg gaacgtcta gtggagaat atttcgccg gaaattgtg catcgggat tttagaatt tggttgacg tcgacgccg tcgccgcat ttcccggtt tcgatttta caaaacggg agtaagatg acgatcgag ccgccggcg gggattaaa gccaacata atgcgtgct tatacgaat caagtgtct gatttgatc gatatcctg cacggcgat ggcccgaaa aatttcaat ttcaagtgg ttattgttc tcgttgtgt ctcttccac ggcgttttg ctcgaaagg cgcaaattc gggccgttg ggattcaat atcccttat gaatttaca gacggcgat ctgaaaatt tgcatctcg caactgagg atgttcctg tgggagtat gacgaaatt cctttcaaa gttttgaca tacacggcg ggtcacatc aattacggg ggaagggtg acggatgac tgggataga aggtgtctg atgaatatt ttagccgat tattacaaa ccggaagtt gtcaatgcg gattacgtc tttgatgcg cacgggttt taccaccaa atgccggcg gaaacgcca tattatgat tacttggat tatatcaaa cattttccg attaacgat gatccggaa ttgttcggg atgcatcct aatgctgat atatcgttc gcacaagca cagacttat agttgcttg tcaactctg ttgacgctc cagccgaga caagttgga ggtgcagct gcgagcgag gaggaagtc attgcacaa acggcagct tctatactt aatcatttg ccgagacaa ttcgatttg gaagcgatt tcggagaaa tatcccgtc ctgtacgaa gaatcttta aacacggtt ttaatccaa gaaggaatc cgctttaat cgcctgctc aaagcaata gaaagcagc ctcaaagac ctgcttaag gccatcaaa gggctcgtc gtaatgtct gaagcactt gataaaatg gcgactagt ttattcagc aacatcgtc ccagccctc tggtctagt aaagcctac ccttcactc aaaccctta ggcgcttgg gtcgccgac ttagacgcc cgtgtcaag tttttaaac acgtgggtc gaccaaggc atcccttca gtcttctgg atttcgggt ttctacttc cctcaagct ttccttact gcgactttg cagaatttc gctcgaaaa tatgtcgta tcgattgat acgattaac ttttcgttt aaggttctc gaacatgcg cctgcaaaa cgaccggat gacggatgt tgtatatgg gggctgttt gtggaaggc gcgaggtgg aacagttcg gtggggact ttagatgag tcgcacccg aaagaatta tatactgaa atgccggtg gtttggtta ctaccggag gaaaatcat gtcaaaccc gaaggcaga tatgagtgt ccggtttat aagactttg acaagggct ggggttttg tcaactacc ggacattcg acgaattat gttttggcg attgaagtg ccgagtaaa aaatcggaa gcgcattgg atcaaaaga ggagtggca ctaatttgt gcactggat tattaa
- >TC007517 RA
tagtgctac gaagtgttg aaagctgcg atgacggcg ctcgatggg cagccccag cccagcggc caaccattc cgtcctgtg cacacctac gtcatgaac cccaaatcc atcacaatg ggccaatta tacggcgag tttgatttg cagactcac gaatggacg gacgggatt ctcccgtgt ttggtgcgc attggagtg gctgcggag aacaaagac aagcgttgg tacgttttc gacgggccc gtggacgct gtctggatc gaaaacatg aataccgtc cttgacgat aataaaaaa ttgtgcttg agcagcggc gaaataatc aaactgcga gatacaatg acgatgatg tttgaagtt gcagattta gctgtagct tctccggcg actgtctcc cgatgcggg atggtttac ctcgagccg ggagttttg ggattagcg ccgtttatc aactgttgg ttgaaacga ctcccaaag ctggcggcg cctttcgcc gacaaattc aaagagttg atttatgca tatgtggtg ccagggatt gaactagtc cgtaaccgg cttagggag atcttgact tctgttgat tcggccctt ttgtacaaa tttctacgt ttgatggat tactggctt ttgccgtta tcggggcgt gataataaa ccgccccca ggaccacag tttttagct ttgattccc gagcttttg gctccttgg gtcattttt tcgatgatt tggagcatc ggctgcact tgcgataac cggagtcgc gtacgtttc gaccattgg ctcagggga aaaatgcta gaggaagga gatgagccg tactttccc cagaaaggt ctggtgtac gattataga ttgcacgat ggtgggttt actgaccct acggaggat ggaaatccc atgccaccg cactggtac agctggatg gacaatgtg gaagaattt aaaatcaca gtggacatg aaatattcc gatatggaa gttccgact atggatact gtacgaaac gccaaaatg ttggaaata gttttgaac aactacgat aatgtgtta tgtgtcggc ccaacggga actggaaaa tctctcaca gttgtcgga aagttgagc cgaagtatg cacaagaag tttatttgc gattttatg agtttctca gcgaggact tcggctaat caaacccag gatttgctc gattcaaaa ctcgatcga cgaagaaaa ggcgtgttt ggacctccg gtcttaaaa cgccagatc tttttcatt gatgatttt aacatgccg gctttagaa gtgtacggt gcccaacca cccatcgaa ctaattcgt caatggatg gattttgga ggttggtat gaccggaaa aacatcggc gactttcgc acaattatt gatacgaat ttttgtgtc gcgatggga ccccctgaa gggggcaga aacccggtc acggctaga ttattgaga catttccat tatttggcc ttcatcgaa atggaggat gatagtaag aggaaaatt ttcggaaca atttttaaa ttttggatg gagcggata ccggaagag tttaccttg gggcatttt tacgaaccg attctggaa gcaacactt aatgtttat agcacgatt ttaaaagaa ctgcttcct acgccagca aaaacgcac tatactttt aatttgcgg gatttgagt aaggttttt caagggttg ctaatgttt aatcctgaa aaaatgacg gacataaat ttattggtg cggttgtgg taccacgag tgttgtcgc attttccaa gaccgatta gtgaacgac gacgacaga aactggttt gcaaatttg ctcaaaaat aaaatcgag agtgagttt gaagttgat cctgtttcg gccttaggg actcagatg ttgtttttt gccgatttt attgatcca acgactgat tatcgcgaa tatgaacaa attactgat tcagcacgg ttatcaaac gtccttgat tactatttg aacgagtat aacgagcag agcacgcgt ccgatgaag ctggtcctt ttcctggat gcaatttcg cacgtttgt cgcatttcg aggatcatt cggcagcca ctaggcaat gcacttctg ttgggtgta ggcggatcc ggacgacag agtctgacg agacttgcg acctttatg gcggagtac gtttgtttc cagattgaa attcacaaa acttacggc atgtttgaa tggcgcgag gacatcaaa ggactaatg cttaaagcc ggtcttcac agacgcgaa actgttttt cttttctca gacacacaa attaaatcc gagtcgttc ctagaagac ttaaataat gtactaaat tccggagac gtcccgaat atttatcaa ccggacgaa ttggataag attttccaa gaaatgaga ggagtcgtg caagaaatg ggactcgtg gcaacaaaa tctaattta tttgccact tatcagaaa gtagtgaga gcaaatttg cacacagtg gtcacaatg agtccgata ggtgatatt ttccgggca agactacga cagtttccg gctttagtc aattgttgc acgatcgac tggtttagc gaatggcca gattcagcc cttaggtcg gtagcgttc caatttttg gatgacatg gctgaatta gacgtagga caagaagtc ttgcatgga attgtcatg gtgtgtcaa ttcatgcat gcaagtgtt gtcgatgcg agcattctt tacaagcaa gagctttcg agacataat tatgtcacg ccaacgtct tatctcgaa cttttgtca agctatact gacctcatg ttgaaaaaa agaggagct ttatcggag ggcattacg agattgtca accggcttg gataagctc aaaagcact gccgaagaa gtaaagatc cttcaggcc agccttgaa gaaatgaaa ccggccttg gaagccgca aaacaagac gccgaaaca atgattgct gaaatcgcc aaagatcaa atcatcgca gaagagacg aaagaaatt gtcgaaaag gaagagctc gaggcggcc aaaaaagct ctcgaaacc caaacaatc gccgatgac gctcaacga gatttagat gaggcaatg ccggatctg ttagcggcc gaagctagt ttaaaatcg cttaataaa aacgatatt tccgaagtg cgatcgatg aagcgtccg ccggccggc gtagtatac gtcattgaa tcgatttgc atcattaaa aacatcaaa ccggcgaaa ttacccgga agaatgccc ggagagaaa gttttagac tactgggag ccgggtcgc aacatgctc gctgatccg ggcgtgttt ctaacgtcg ttaatgaac tttgacaaa gaatcaatc accgaggaa atgatcgac aagctgaaa aattacgta tcgaatcca caatttcag cctgcaaaa gtggccaag gtctctaaa gcgtgcaaa tcgttctgt atgtgggtt catgcaatg tataaatac tactttgta aataagcga gtggcgccg aagaaggcc gccttggca caggcgaaa gccgagttg gccaaaacc gaagccgct ttagccgca gccaaggca aaaatgcaa gcggttttg gataattta agattgttg caagaaaaa ttaggggcg aaaattgct ttcaaagag gagaaagaa gcgagtatt caaatttgt ttggaacgg atgagtcga gcagttcgc ctaatcaat ggattatcc gatgagaga gttcgctgg ttgaacacc atttcgagc attgaagca agtgttgtc aatgtcacg ggtgatata ctcctgagt gccggtgct gtggcttat ctcacccca tttacggat aaatacaga cgagggttg ttgtccgaa tggctcaaa gagataaag gagcaaaag ataccctgt tcggctgat cctaatccg gtttcggtt ttagccgaa ccggtccag atacgcgca tggcaattg gacggctta ccgagggat tatttctca acggagaac gctgttctc gtttcgaat tcgaaacgt tggcctttg tttatagat ccacaggga caggccaat aaatgggtt aagaatatg gaaagggct caaggattg gcggtgtgt aaaatggcc gataaggag ctgttgcgt actttagag tcggcggtt aggtttggt aaaccagtt ttgattgag ggagtcgga attgattta gatccgtct ctggatcca gttttactg cgccaaaaa tttaaacag gcaggaacg tggatgttg aagctgggt gaagtcgtc gtgccttac gatgacgat tttcattta tacatgaca acgaaatta ccgaatccg cattatacg cctgaagtt tcagttaag gttcttttg gtgaatttt actctggtc ccgagtggc ttgcaggac cagctgctg gctttagtt gtcatgcag gagcgtccc gatttggag gaccagaga agtcagttg attgtcgga agtacacag atgaaacag gaattgaag gaaattgaa gacaggatt ttgtacaag ttgagttca ttggaaggt tcacctttg gatgattta gattttata ataacactc gaagcatcg aaagttaag agcgacgat attaaaaat aaagtggtc gctgctgaa ataacacaa atcgacatc gacaataca cgtgccttg tacattccc gtagcaaat agggcacag atattgttt ttctgtttg tccgattta tcaaacgtc gaccctatg tatcaatac tcgttggag tggttcgtt ggtattttc atagcaagc atggccgag acggaacga accgaaaat attgatgaa agagttgtc ataatcaac gaatacttt acgttcagt ttgtatagt aatgtttgt cgcagtttg tttgaaaaa cacaaatta cattttgcg tttttgatg tgcgcccgg atatttatg gacatgggg aaaattgac gggcacgaa tggcatcac tttttggca ggaggcacc cctctctcg gaagatcca aatcccgcg cctgattgg ctctcgggc aaagcatgg aaggaaatc ctcgcaatg cgtgtcctt ccaaattac gttcgattc gtcgatact ttcgcacaa tacaaagaa gattatcgc gtcatattt gagtctggg gaccctcac agaaaaccg ttacccgaa gagttcgaa aaatcattc tccgagttt caaaagcta ttagttttg aagtgtttg cgtcccgac aaagtgaca aacgccatg caagatttc atttcgtcc aaaatgggt cagagattt atcgaacca cagacgacc gacttatcc gtcgtcttc aaagactcc gggcctaca acccctttg attttcgtc ctgtccact ggaacggat ccggccgca gatctgtac aagttcgca gacaagatg aaattcgcc aagagaatg ttttcgatc tcgctgggc caaggccaa ggacccatt gccgagaaa atgctacac aacgggact gaactcggc tcgtggatt ttcttccag aactgtcac ctggcgccc agttggatg ccgcggttg gaacgtcta gtggagaat atttcgccg gaaattgtg catcgggat tttagaatt tggttgacg tcgacgccg tcgccgcat ttcccggtt tcgatttta caaaacggg agtaagatg acgatcgag ccgccggcg gggattaaa gccaacata atgcgtgct tatacgaat caagtgtct gatttgatc gatatcctg cacggcgat ggcccgaaa aatttcaat ttcaagtgg ttattgttc tcgttgtgt ctcttccac ggcgttttg ctcgaaagg cgcaaattc gggccgttg ggattcaat atcccttat gaatttaca gacggcgat ctgaaaatt tgcatctcg caactgagg atgttcctg tgggagtat gacgaaatt cctttcaaa gttttgaca tacacggcg ggtcacatc aattacggg ggaagggtg acggatgac tgggataga aggtgtctg atgaatatt ttagccgat tattacaaa ccggaagtt gtcaatgcg gattacgtc tttgatgcg cacgggttt taccaccaa atgccggcg gaaacgcca tattatgat tacttggat tatatcaaa cattttccg attaacgat gatccggaa ttgttcggg atgcatcct aatgctgat atatcgttc gcacaagca cagacttat agttgcttg tcaactctg ttgacgctc cagccgaga caagttgga ggtgcagct gcgagcgag gaggaagtc attgcacaa acggcagct tctatactt aatcatttg ccgagacaa ttcgatttg gaagcgatt tcggagaaa tatcccgtc ctgtacgaa gaatcttta aacacggtt ttaatccaa gaaggaatc cgctttaat cgcctgctc aaagcaata gaaagcagc ctcaaagac ctgcttaag gccatcaaa gggctcgtc gtaatgtct gaagcactt gataaaatg gcgactagt ttattcagc aacatcgtc ccagccctc tggtctagt aaagcctac ccttcactc aaaccctta ggcgcttgg gtcgccgac ttagacgcc cgtgtcaag tttttaaac acgtgggtc gaccaaggc atcccttca gtcttctgg atttcgggt ttctacttc cctcaagct ttccttact gcgactttg cagaatttc gctcgaaaa tatgtcgta tcgattgat acgattaac ttttcgttt aaggttctc gaacatgcg cctgcaaaa cgaccggat gacggatgt tgtatatgg gggctgttt gtggaaggc gcgaggtgg aacagttcg gtggggact ttagatgag tcgcacccg aaagaatta tatactgaa atgccggtg gtttggtta ctaccggag gaaaatcat gtcaaaccc gaaggcaga tatgagtgt ccggtttat aagactttg acaagggct ggggttttg tcaactacc ggacattcg acgaattat gttttggcg attgaagtg ccgagtaaa aaatcggaa gcgcattgg atcaaaaga ggagtggca ctaatttgt gcactggat tattaaaaa aaacattta ttttaagtt ttttttatt aaattaatt aaagatatt tttgtaaca aacttggta acaaagatt tgtgattat aactgtttt ctaaagcgc aagctagga atctttgaa cgttcaaaa tttcgccgg aaataacat taacaaact tcctctctt taccatgag taaagagaa gacgtttgc ttatgtagt caccataca atgacaagt aaccaactt ctacatatg ttcctgaat aattcgaaa acatttcta ttgaaactt tcaaactct cgcccactt ttaaatcaa cagcgggag cgtaaacta atcgaaagt gataattca gaggttcat aaatttccc gaatttgtt gtaccagtg aatcaaaaa tttcattgc tgttgttgc tagtagcta caaaaattt gcccgactg tattttggc tcaaattaa gtaaatcgg ggccacttc cagggctgt atttgcgcc ccagtgtga aaaatttca gaggcaaca gagccgact gtattgaat ttttggcga aatatgcca tgaaagaat acaaagagg cgcccccga ctaaatgaa gccgattga tctcgttat ttgagccgt ccaaagtga aaattttat ttttatcca agtaatcgg tcccgcaac cttcgacaa caagactac ggacaaaat cagcgtttg aaaattggg tgaagcttt ggcttacta aatttgatt ggaaataat ttgttatgg caaattcga aaagagcgg cgtctgatt tcaaataac agagagtcg ggcttgtgt attcaatta aattttggg atttagcga tttggaggt gattttctt gaatcggtt taggctgct ctaaaaact catgaatga ttgtttgag aatcaaggg gtgttttag ggtgtaaga aattaggta acgagagaa cacccaagt ttgcgttct cttcgagcc cgtaggtaa attctttca cgtttttta aatcgtctt ttactactt taatatgaa gcaataatt tttctatgt cactctgat ttttttcct ccccgtcct gtttcaatt tatttatca agttttgta tttctgatc ttgtgaatt caaggacgg ttttgtggt catcaattg tcttaaaaa aattccaac gttgttcaa ttttatcca agttaatcg tcagcttac gcgcttcta tacttttaa ttaaatcat ttttgtttt ttagtattt cgtcaaaat caacaaaag gcgtgagaa caacaaaat gatgttgcg ctttatcac ctgttacgt acaaagccg acgaaaacc ggcgcacag tttttgtaa acacttgca caagtcttg ccatgttat gcaattttc attttcttg ctttctgag acagcttcc gggtcaatt gtttcaatt ttgcgtttc tccggtgct ttccagtta ccaaaaccg gcagctctc tcggattct tctatggtg tagggctcc aactccagg gctttcagt ctgcgtttg tggactttg gttctgaag tgttcgtgc agagcgtgg tcatcaata aaataacgc gcacaatgg acgcaataa aactgggca ctgcccggc ttatcgtag tctacttct tggttcacc aacttttcg gcattttcg ggcttgatg tcctcatcg atctgtacc aacacgtga gcctgtttt ggggttagg actattatg acattacct gatccaaat ctttggttc ttcgcttcg tacgccact ttttcttca agtgcgtat cgccataat ggtactttt tgcgtttat acaccattt aaaaataga tttaatgca aaattgaac actttttgg cagttggaa gtggttgag gttgcgtca attgtggtt gcgtgt
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