| General information | |||
| TC008263 | 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: 7487406..7498377 | |
| 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 |
|
TC008263 | |
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_02771 | 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: 100.0% (includes death as pupa, adult)
eclosion not fulfilled (with pupal and adult features) - Number of animals showing the phenotype: 2 (3 dpi)Days post injection
adult lethality - Number of animals showing the phenotype: 8 (9 dpi)
Days post injection
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: -1.0% (includes death as larva, prepupa, pupa)
Lethalities 22 days after larval injection: -1.0% (includes death as larva, prepupa, pupa, adult)
- >TC008263 RA
MILTEATMN PEEIKAEYT SSLQDLTFN SKPLINVLT MLAEENLAH APYIVQAIE EHLEKVQTN VKLPILYLI DCIAKNIGQ TYMQLFSQN IVKTFCSVF KVVDEETRA EMFKLRQTW NNVFPQMKL YAIDVQIRM LDPAWPVTA QPTSNSIHF NPKFLKTTK TEIATPIVP ETTTTPLDM ETFTMQQKL IQKQKELIE LQQKKLKLE LLQTQVKLQ KQLENNPNA VVGNNVPPR TQTGTGKTY TMIGNQAIN PEDSTIGLI PRAAAHLLE ELEQINSKI EYTVRISFI EIYNEEVHD LLNSSSIPL KIFEDPDNK GSVCIKGVK EATVLNLQE VYDWLNIGL VERQTASTN MNRHSSRSH SIFTISVLT RQLTVEGDE LITIGKLYL VDLAGSENV GRSGSTELR AREAGNINK SLLTLGKVI KALAQKTQH VPYRDSKLT RILQDSLGG KTKTCMIAT ISAAANVWE ETVSTLDYA QVARNVSNC PQVNENRKQ ANILKDLKD EISRLRKEL AIARSGEGF YTSRENHER LCADVERLQ TEVNEEKRK LEQVTKKNS ELELAISEK DEKLRNVSQ YCDDLKREL REKDLQEKR NKSVIDYFE TREKEMRQA AEQLRKVCE VSTKHETLF CQKYENQRA ISLKNATAA KNAYNLFGD SLRALQNAT EDHIQRQNV SLSEVQANV GCLKENAWK MCKRLTEYS AKMQDDSQG DFYKLDESN LSRDFEQRL ESMFTSQFF AIQKKAVQD LEQYNEEKM HEVETMTTR CINSMKQSL NDYMLCVND IMKNQSSLI ASVKGATNS DEVNILLKP EVAKQLMPE VAKQLIPAA TRDTKSKSG LGTRAQQTT PKITPVNSA LVAAATKPT RDPRLLRQQ TEKTTNGTN NSSSTSVSL GSKSSILDS NNKVVTNTK SVRDNRIES RLVTNKDNA PPPKLDKSK TFLKSRQSE QKPNKPPRS KNSADNDSK PFSGKSSIN LLDSPVKIK SDLKSPTKS PNRHKKKDH STDKKGFKD AKRDRDNKG HKNDSLQTT FKGVKAKNR NYIRRNRSP SYSPEPNQD TDLRVSGPP EKQARLHPE TVEEKSKLV AQTGMDVDL RQLNDVTKK RASSEGLEN SNKKSKTEV FDALFGNED TDLRTVLPV AQPERPPTP PPPIISLTD KEDSPKKSS DFDAVRAKL ANATGRDKI MAKSFRKKR TQDEDLRVP PVSSRISDT IKKIIISSD DAQCIKTGT MSKEKENQL MNKIIAHIE QQKLEEAQL RDSEETVGN LQPVSDEEL EFSEETDEK FGDKDDRVN FHKYSRDSR ESRRGGSYW RGGKRRHWE NPHMGPHPR SAIRPWLHQ MHAWRPIGP NFNRQVHTP EHDFSSQEP DGKKSPETA VEVTVDSVN QDDIKTINI DAHPRDIRY YDETAIVFM NWDDPREIS FQNGSRRVT FDDKDTYLL SFNEGYKEL CINDTIHKV RLGAPSREI FIDGKPYEC YFGGPGITI DLNGKMTVV KLDGPPPQV KIGTVKRTD LVAGKVNLI IDAKILVPV FLDAKVQKF VIDGETHTL KFVDALKTV LINDVPFKV EFGGLPKPI TVHDKKHFI RFSVLPRGV KPGLVCIKD MEGVQAPET DESQDTALA VDINEPALP VFSRKKKVE DEQDRNSNS PHFFQSFMQ QNNLDVLTS VMTSSMTPS ATSGGYQVE NVGPNTETH SMDAIPYVK PAPVLPSSL NINELFQKL VATGIVATQ EPVAEPQPP PPVKTVRRS TMENLRPVT FNRPETLKM RQSLLYEAL YSGMQCSSC GMRFPPEQS IHYSQHLDW HFRQNRKGK KNIRVAASR RWYYSLSDW KNYEEIEDL EEREKNYFD QQQLQAEGA GDDVDEEVE VPSVPADPD VQDAHCEVC QDRFDHFFN EEKEEWHLR NAVRVEDKT YHPVCYEDY QASLLEPAE ECKMEEPEP EETKIEEAN IPGLEIILD DDDEEEEEE GKFKDEEVI IEESDESKP SNELKEENE NEEDGEDDD VIIHEVVPE KIEVDDDKE YDTLPPSDV PIKVEQIDD GFMDVEEGL IKIKNGGQI KIKSEPIDP DEEMLEVPR EESSVPFTE PVQPSFTSS IDGNTEPVS SVPTTTANI GSKIKINIS KPLPVIAPK EPKDISCDS VPSLEPVID PSQPLPPGE EPVQLNIKP ALRGIELKK LPPVQKGSE LTGMCSIM
- >TC008263 RA
atgattttg acggaggcc acgatgaat cctgaggag atcaaggcg gagtacacc tccagcctc caagatctc actttcaac agtaagccc ctgatcaat gtgttgacg atgttagcg gaagaaaat ctagcacac gctccatac atcgtccag gcaatcgaa gagcatctg gaaaaggtt cagaccaat gtcaaactt cctattctg tatctcatc gactgtatc gccaagaat attggacaa acttatatg caactcttc agccaaaac atcgtcaag acattctgc agcgttttt aaagtggtg gacgaagaa acgcgtgct gaaatgttt aaacttcgt caaacgtgg aataacgtc tttcctcaa atgaaattg tacgcgatc gacgtccag attcggatg ttagatcca gcatggcca gttacagcc caacctaca tccaacagt attcatttc aatcccaaa tttctaaaa accaccaaa accgaaata gcaacacct atcgtccct gaaacaaca accactcct ctggatatg gaaaccttc acgatgcaa caaaaacta atccagaaa caaaaagaa ctcatcgaa cttcagcag aaaaaatta aaattagaa ctgttacaa acacaagtt aaactccaa aagcaattg gaaaataat ccgaatgct gttgttggc aataatgta ccaccaaga actcagacc gggacggga aaaacttac acaatgatc gggaatcaa gcaataaat cctgaagat tcaacaatt ggtttaata cccagggcc gcagctcat ttattggaa gaattggaa caaatcaat tcgaaaatt gagtacact gttcgcatt tcctttatc gagatttac aacgaggaa gtgcacgat cttttaaac agttcatca atcccgttg aaaatattt gaagatccg gataacaaa ggatcagtt tgtatcaaa ggtgttaaa gaggcgact gttcttaat ttgcaagaa gtttacgat tggcttaat attggtttg gttgaacga caaacagca tcgaccaat atgaatcgt cacagctct cggtcccat tcaatattt acaatatcg gttctaact cgccaatta acagtagaa ggagatgaa ttaattacg atcggtaaa ctctatttg gttgattta gccggtagt gagaatgta gggcggtcg ggttctacg gaactgaga gcacgagaa gccggcaac ataaacaaa tcacttctg acattggga aaagtgatc aaagccttg gctcagaaa actcagcac gtaccgtat cgagactca aagttgacc agaattctg caagatagt ttgggaggt aaaacaaag acgtgtatg attgcgacg atttcggca gccgcgaac gtgtgggaa gaaaccgta agcacgttg gattacgct caagtggca cgaaatgtt tccaattgt ccccaagtg aacgaaaac agaaaacaa gccaatatt ctcaaggat ctaaaggac gagattagc agacttcga aaagaactg gctatagcc aggtcgggc gaaggattc tacacaagc cgagaaaat cacgaaaga ttatgtgca gacgtggag agattacaa accgaagtc aatgaggaa aaacgaaag cttgaacaa gtcacgaag aaaaacagc gagctggaa ttggcgatt agtgaaaaa gacgagaaa ttacgaaat gttagtcaa tattgcgac gatttgaaa cgtgagttg agagaaaaa gacttgcaa gagaagcga aacaaaagc gtgattgat tattttgaa acgcgcgaa aaagaaatg agacaagcg gctgaacag ttacggaaa gtatgtgaa gtttcgacg aaacacgaa actttgttc tgccagaag tatgaaaat cagcgcgcg attagtttg aaaaacgcc acagcggcc aaaaatgca tataattta tttggtgac tcgttgagg gctttgcag aacgcgacg gaagatcat attcagcgt cagaatgtc tcgctgagt gaggtacaa gcgaatgtc gggtgtttg aaggaaaac gcctggaaa atgtgcaag cgtttgact gagtattcg gcaaaaatg caagatgat tcacaaggc gatttttat aaattggat gaaagtaat ctttcgcgt gattttgaa caacgtttg gagtccatg tttacgtcc cagtttttt gcaattcaa aagaaagcg gttcaagac ttggaacag tataatgaa gaaaaaatg cacgaggtg gagacgatg acaacgcgt tgtataaat tcaatgaag cagtcgctg aacgattat atgctttgt gttaacgat attatgaaa aaccagtcg tcgttgatt gcaagtgtt aaaggcgca acgaatagt gatgaagtg aatatttta ttaaaaccc gaggtagca aagcaacta atgccagaa gtggcgaag cagttaatt cctgccgca acacgtgat accaaaagc aaatcaggt ttgggtacg agagcccag caaaccacg cctaaaata acaccagta aatagcgct ttagtggcg gctgcaaca aaaccaacg cgtgatcct cgtttattg cgccaacaa acggagaag actaccaac ggcaccaac aacagtagt agtactagt gtttcatta ggatctaag agttcaata ttagattct aataataaa gttgttaca aatacaaag agcgttcga gacaaccgt atcgaatct cgtttagtt actaataaa gacaatgct cctccaccc aaactagac aaatccaaa accttccta aagtccaga caatccgaa caaaaacca aacaaacca ccacgtagc aaaaacagt gcagacaac gattcgaaa cctttctcg ggtaaaagt tcgatcaat ctgttagat tcacctgtt aagattaaa agtgacttg aaatcgccg accaaatca cctaatagg cacaaaaag aaagatcat agtacggat aaaaagggt ttcaaagac gctaagcgt gacagggac aataagggt cataaaaat gactcgttg caaactact ttcaaaggt gttaaggcc aaaaatcgg aattatatt agaagaaat cgctcgcct tcgtacagt cccgaaccc aatcaagac accgacttg agagtatca ggacctcca gaaaagcaa gcacgacta catccagaa actgtcgaa gaaaagagc aaattagtc gcacaaacg ggaatggac gtggattta agacagttg aacgacgtg acaaagaaa agagcttcg agtgaaggt ctggagaac tcaaacaag aaatcgaaa acggaagta tttgatgct ttgttcgga aacgaagat acggatctt cgcactgtt ctaccggtg gcccagcca gaacgaccc ccaactcct ccaccgcca atcatctct ctcaccgac aaagaagat tcgccaaag aaatcttca gactttgat gccgttcga gcgaaacta gccaatgcg accggacgc gataaaata atggccaag tccttccga aaaaagcga actcaggac gaagatctc cgagtacct ccagtgtca agcagaatc agcgacacc atcaagaaa atcatcatt tcttcggac gatgcgcag tgcatcaaa actggaaca atgtcgaag gagaaagaa aatcaactt atgaacaag atcatcgca cacattgag caacagaag ctggaagaa gcacaattg agggatagt gaagagacg gttggtaac cttcaacca gtcagcgac gaagagctc gaatttagc gaagagact gacgaaaaa ttcggggat aaggacgat agagtcaat ttccataaa tattccagg gattcccgg gagtctaga aggggtggg tcgtattgg agaggcggc aagagacgc cattgggaa aatcctcac atgggacca cacccacga tctgcaata aggccttgg ttgcatcaa atgcacgcc tggaggcca attgggccc aatttcaac cgccaagtg cacacccct gaacacgat ttttcaagc caagagcct gatggaaag aaatcaccg gaaactgca gtggaagta actgtggat tcagtaaat caggacgat attaagacc atcaatatc gatgcgcat ccgcgtgat ataagatac tacgacgag actgcgatc gtatttatg aattgggac gatccacga gagatttcg ttccaaaac ggcagtcgt cgtgtgacg ttcgacgat aaagatact tatttattg tccttcaat gaaggctat aaagaactg tgtattaac gacacgata cataaagtg aggttgggg gccccaagt cgtgagatc ttcatcgat gggaaaccc tacgagtgt tacttcggg ggtcccgga ataacgata gatttaaac gggaaaatg accgtagtt aaattagat gggccccca cctcaggtg aaaatcggc accgtgaaa agaaccgat ctggttgcc ggcaaagta aatctgata atcgatgcg aaaatctta gttcccgtt tttctggat gccaaagtg caaaagttt gttatcgat ggtgaaaca catacatta aaatttgta gatgctttg aaaactgtt ttaataaat gatgtacca tttaaagtt gaatttggt ggcctccct aaacctata actgtccat gataagaaa cactttatc agattttca gtgttgccg aggggtgtt aaaccaggt cttgtgtgt attaaagac atggaaggg gtgcaagcg cctgagact gacgaaagt caggacaca gcgcttgcc gttgacatt aacgagcct gctctgccc gtcttctcc cgcaagaaa aaggtggaa gatgagcaa gacaggaac tcgaactcg ccacatttc ttccagagt ttcatgcag caaaataat ttggacgtg cttacaagt gtgatgact tcctcgatg acaccaagt gccaccagt ggaggttac caagttgaa aatgtgggg ccgaacact gagactcat tccatggac gccatcccc tacgtgaaa ccagcaccg gtcttgccg tcctcgctc aacatcaac gaactgttc caaaagttg gtagcgacg ggaatcgtc gcgacccaa gaaccggta gccgaacca caaccacca ccacctgtt aaaactgtg agacgcagc acgatggaa aacctcaga cctgttacc ttcaacaga cctgaaacg ctgaaaatg cgccaaagt cttctgtac gaagcactt tactcaggg atgcagtgc agcagttgc ggcatgcgt ttcccgcca gagcagagt atacactac agccaacac ttggactgg catttccga caaaaccgg aaagggaaa aagaacatc agagttgcg gcgagtcgc agatggtac tattcactc tccgattgg aagaattac gaagaaatt gaagatttg gaggaacgg gagaagaac tatttcgac caacagcag ctgcaagct gagggtgct ggtgatgat gttgatgaa gaagttgaa gtgccaagt gttccagcg gatcctgac gtacaagac gcccattgt gaagtgtgc caagacaga ttcgaccat ttcttcaac gaagagaag gaagaatgg catttgaga aatgcggtt agagttgaa gataagact tatcatcct gtttgctac gaagattat caagcatcg ttgctagaa ccggctgaa gaatgtaaa atggaagag ccggaaccg gaagaaact aaaattgaa gaagcgaat atccctgga cttgaaatc attttggac gatgacgac gaggaagaa gaagaagaa gggaagttt aaggacgaa gaagttata attgaagaa agcgacgag tcgaagcca tcgaacgaa ttgaaagaa gaaaatgaa aatgaagaa gatggcgaa gatgatgat gtaatcatc catgaagtt gtacctgag aaaattgaa gtggacgat gacaaagaa tatgacact ttaccaccg tccgatgtg cccatcaag gtggagcaa attgatgat ggttttatg gacgttgaa gaaggccta attaaaatt aaaaacggg ggacaaatt aaaattaaa agtgaaccg attgatccc gatgaagaa atgttagag gtgccccga gaagagtca agcgtgcca tttacagag ccggttcag ccctcgttt acaagttcg attgatggt aataccgag cccgtttcg agtgttccg acgacgaca gcaaatatc ggaagtaaa attaaaata aatataagt aaaccttta cctgtgatc gccccaaaa gagccgaaa gatatttcg tgtgatagc gtgcctagt ttagaaccg gttatagat ccatcacag cctctaccg ccgggggaa gaaccagtc caattaaat atcaaacca gctctgaga ggcatagaa ctgaagaaa ttaccgcca gtgcaaaaa ggaagtgaa ctgacggga atgtgtagt ataatgtga
- >TC008263 RA
aagtgaaaa aacgcaacg ttctgcgtc cgccatatt tgtttgaga gtgtgtgtt gtgtgttaa ttggacttt tctgcagtt taaactaat caaatttgg ttttatgca acatttttt agatgaata cgcttaagg gattaatgt tttactctt caaggacac tcacatgat tttgacgga ggccacgat gaatcctga ggagatcaa ggcggagta cacctccag cctccaaga tctcacttt caacagtaa gcccctgat caatgtgtt gacgatgtt agcggaaga aaatctagc acacgctcc atacatcgt ccaggcaat cgaagagca tctggaaaa ggttcagac caatgtcaa acttcctat tctgtatct catcgactg tatcgccaa gaatattgg acaaactta tatgcaact cttcagcca aaacatcgt caagacatt ctgcagcgt ttttaaagt ggtggacga agaaacgcg tgctgaaat gtttaaact tcgtcaaac gtggaataa cgtctttcc tcaaatgaa attgtacgc gatcgacgt ccagattcg gatgttaga tccagcatg gccagttac agcccaacc tacatccaa cagtattca tttcaatcc caaatttct aaaaaccac caaaaccga aatagcaac acctatcgt ccctgaaac aacaaccac tcctctgga tatggaaac cttcacgat gcaacaaaa actaatcca gaaacaaaa agaactcat cgaacttca gcagaaaaa attaaaatt agaactgtt acaaacaca agttaaact ccaaaagca attggaaaa taatccgaa tgctgttgt tggcaataa tgtaccacc aagaactca gaccgggac gggaaaaac ttacacaat gatcgggaa tcaagcaat aaatcctga agattcaac aattggttt aatacccag ggccgcagc tcatttatt ggaagaatt ggaacaaat caattcgaa aattgagta cactgttcg catttcctt tatcgagat ttacaacga ggaagtgca cgatctttt aaacagttc atcaatccc gttgaaaat atttgaaga tccggataa caaaggatc agtttgtat caaaggtgt taaagaggc gactgttct taatttgca agaagttta cgattggct taatattgg tttggttga acgacaaac agcatcgac caatatgaa tcgtcacag ctctcggtc ccattcaat atttacaat atcggttct aactcgcca attaacagt agaaggaga tgaattaat tacgatcgg taaactcta tttggttga tttagccgg tagtgagaa tgtagggcg gtcgggttc tacggaact gagagcacg agaagccgg caacataaa caaatcact tctgacatt gggaaaagt gatcaaagc cttggctca gaaaactca gcacgtacc gtatcgaga ctcaaagtt gaccagaat tctgcaaga tagtttggg aggtaaaac aaagacgtg tatgattgc gacgatttc ggcagccgc gaacgtgtg ggaagaaac cgtaagcac gttggatta cgctcaagt ggcacgaaa tgtttccaa ttgtcccca agtgaacga aaacagaaa acaagccaa tattctcaa ggatctaaa ggacgagat tagcagact tcgaaaaga actggctat agccaggtc gggcgaagg attctacac aagccgaga aaatcacga aagattatg tgcagacgt ggagagatt acaaaccga agtcaatga ggaaaaacg aaagcttga acaagtcac gaagaaaaa cagcgagct ggaattggc gattagtga aaaagacga gaaattacg aaatgttag tcaatattg cgacgattt gaaacgtga gttgagaga aaaagactt gcaagagaa gcgaaacaa aagcgtgat tgattattt tgaaacgcg cgaaaaaga aatgagaca agcggctga acagttacg gaaagtatg tgaagtttc gacgaaaca cgaaacttt gttctgcca gaagtatga aaatcagcg cgcgattag tttgaaaaa cgccacagc ggccaaaaa tgcatataa tttatttgg tgactcgtt gagggcttt gcagaacgc gacggaaga tcatattca gcgtcagaa tgtctcgct gagtgaggt acaagcgaa tgtcgggtg tttgaagga aaacgcctg gaaaatgtg caagcgttt gactgagta ttcggcaaa aatgcaaga tgattcaca aggcgattt ttataaatt ggatgaaag taatctttc gcgtgattt tgaacaacg tttggagtc catgtttac gtcccagtt ttttgcaat tcaaaagaa agcggttca agacttgga acagtataa tgaagaaaa aatgcacga ggtggagac gatgacaac gcgttgtat aaattcaat gaagcagtc gctgaacga ttatatgct ttgtgttaa cgatattat gaaaaacca gtcgtcgtt gattgcaag tgttaaagg cgcaacgaa tagtgatga agtgaatat tttattaaa acccgaggt agcaaagca actaatgcc agaagtggc gaagcagtt aattcctgc cgcaacacg tgataccaa aagcaaatc aggtttggg tacgagagc ccagcaaac cacgcctaa aataacacc agtaaatag cgctttagt ggcggctgc aacaaaacc aacgcgtga tcctcgttt attgcgcca acaaacgga gaagactac caacggcac caacaacag tagtagtac tagtgtttc attaggatc taagagttc aatattaga ttctaataa taaagttgt tacaaatac aaagagcgt tcgagacaa ccgtatcga atctcgttt agttactaa taaagacaa tgctcctcc acccaaact agacaaatc caaaacctt cctaaagtc cagacaatc cgaacaaaa accaaacaa accaccacg tagcaaaaa cagtgcaga caacgattc gaaaccttt ctcgggtaa aagttcgat caatctgtt agattcacc tgttaagat taaaagtga cttgaaatc gccgaccaa atcacctaa taggcacaa aaagaaaga tcatagtac ggataaaaa gggtttcaa agacgctaa gcgtgacag ggacaataa gggtcataa aaatgactc gttgcaaac tactttcaa aggtgttaa ggccaaaaa tcggaatta tattagaag aaatcgctc gccttcgta cagtcccga acccaatca agacaccga cttgagagt atcaggacc tccagaaaa gcaagcacg actacatcc agaaactgt cgaagaaaa gagcaaatt agtcgcaca aacgggaat ggacgtgga tttaagaca gttgaacga cgtgacaaa gaaaagagc ttcgagtga aggtctgga gaactcaaa caagaaatc gaaaacgga agtatttga tgctttgtt cggaaacga agatacgga tcttcgcac tgttctacc ggtggccca gccagaacg acccccaac tcctccacc gccaatcat ctctctcac cgacaaaga agattcgcc aaagaaatc ttcagactt tgatgccgt tcgagcgaa actagccaa tgcgaccgg acgcgataa aataatggc caagtcctt ccgaaaaaa gcgaactca ggacgaaga tctccgagt acctccagt gtcaagcag aatcagcga caccatcaa gaaaatcat catttcttc ggacgatgc gcagtgcat caaaactgg aacaatgtc gaaggagaa agaaaatca acttatgaa caagatcat cgcacacat tgagcaaca gaagctgga agaagcaca attgaggga tagtgaaga gacggttgg taaccttca accagtcag cgacgaaga gctcgaatt tagcgaaga gactgacga aaaattcgg ggataagga cgatagagt caatttcca taaatattc cagggattc ccgggagtc tagaagggg tgggtcgta ttggagagg cggcaagag acgccattg ggaaaatcc tcacatggg accacaccc acgatctgc aataaggcc ttggttgca tcaaatgca cgcctggag gccaattgg gcccaattt caaccgcca agtgcacac ccctgaaca cgatttttc aagccaaga gcctgatgg aaagaaatc accggaaac tgcagtgga agtaactgt ggattcagt aaatcagga cgatattaa gaccatcaa tatcgatgc gcatccgcg tgatataag atactacga cgagactgc gatcgtatt tatgaattg ggacgatcc acgagagat ttcgttcca aaacggcag tcgtcgtgt gacgttcga cgataaaga tacttattt attgtcctt caatgaagg ctataaaga actgtgtat taacgacac gatacataa agtgaggtt gggggcccc aagtcgtga gatcttcat cgatgggaa accctacga gtgttactt cgggggtcc cggaataac gatagattt aaacgggaa aatgaccgt agttaaatt agatgggcc cccacctca ggtgaaaat cggcaccgt gaaaagaac cgatctggt tgccggcaa agtaaatct gataatcga tgcgaaaat cttagttcc cgtttttct ggatgccaa agtgcaaaa gtttgttat cgatggtga aacacatac attaaaatt tgtagatgc tttgaaaac tgttttaat aaatgatgt accatttaa agttgaatt tggtggcct ccctaaacc tataactgt ccatgataa gaaacactt tatcagatt ttcagtgtt gccgagggg tgttaaacc aggtcttgt gtgtattaa agacatgga aggggtgca agcgcctga gactgacga aagtcagga cacagcgct tgccgttga cattaacga gcctgctct gcccgtctt ctcccgcaa gaaaaaggt ggaagatga gcaagacag gaactcgaa ctcgccaca tttcttcca gagtttcat gcagcaaaa taatttgga cgtgcttac aagtgtgat gacttcctc gatgacacc aagtgccac cagtggagg ttaccaagt tgaaaatgt ggggccgaa cactgagac tcattccat ggacgccat cccctacgt gaaaccagc accggtctt gccgtcctc gctcaacat caacgaact gttccaaaa gttggtagc gacgggaat cgtcgcgac ccaagaacc ggtagccga accacaacc accaccacc tgttaaaac tgtgagacg cagcacgat ggaaaacct cagacctgt taccttcaa cagacctga aacgctgaa aatgcgcca aagtcttct gtacgaagc actttactc agggatgca gtgcagcag ttgcggcat gcgtttccc gccagagca gagtataca ctacagcca acacttgga ctggcattt ccgacaaaa ccggaaagg gaaaaagaa catcagagt tgcggcgag tcgcagatg gtactattc actctccga ttggaagaa ttacgaaga aattgaaga tttggagga acgggagaa gaactattt cgaccaaca gcagctgca agctgaggg tgctggtga tgatgttga tgaagaagt tgaagtgcc aagtgttcc agcggatcc tgacgtaca agacgccca ttgtgaagt gtgccaaga cagattcga ccatttctt caacgaaga gaaggaaga atggcattt gagaaatgc ggttagagt tgaagataa gacttatca tcctgtttg ctacgaaga ttatcaagc atcgttgct agaaccggc tgaagaatg taaaatgga agagccgga accggaaga aactaaaat tgaagaagc gaatatccc tggacttga aatcatttt ggacgatga cgacgagga agaagaaga agaagggaa gtttaagga cgaagaagt tataattga agaaagcga cgagtcgaa gccatcgaa cgaattgaa agaagaaaa tgaaaatga agaagatgg cgaagatga tgatgtaat catccatga agttgtacc tgagaaaat tgaagtgga cgatgacaa agaatatga cactttacc accgtccga tgtgcccat caaggtgga gcaaattga tgatggttt tatggacgt tgaagaagg cctaattaa aattaaaaa cgggggaca aattaaaat taaaagtga accgattga tcccgatga agaaatgtt agaggtgcc ccgagaaga gtcaagcgt gccatttac agagccggt tcagccctc gtttacaag ttcgattga tggtaatac cgagcccgt ttcgagtgt tccgacgac gacagcaaa tatcggaag taaaattaa aataaatat aagtaaacc tttacctgt gatcgcccc aaaagagcc gaaagatat ttcgtgtga tagcgtgcc tagtttaga accggttat agatccatc acagcctct accgccggg ggaagaacc agtccaatt aaatatcaa accagctct gagaggcat agaactgaa gaaattacc gccagtgca aaaaggaag tgaactgac gggaatgtg tagtataat gtgattaat ttaaatagt gctgagtga gtgtgtgta catattgca gactaaaag ggggactgt ttatataaa attcgtttc gttttatca tctgcacta aaatggatt ttttacgat ttattttca ctctgtacc tattatgcg tatcatgat ttagtttga cattttatg tctctgttg ttgtcaatt gttggaaat aaatttatt ttttaaaa
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