| General information | |||
| TC015351 | 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_007421.3: 102777..110074 | |
| 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 | TC015351 | ||
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_05800 | 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: 30.0% (includes death as pupa, adult)
pupa eclosion not fulfilled (died as pupa) - Number of animals showing the phenotype: 1 (3 dpi)Days post injection
eclosion not fulfilled (with pupal and adult features) - Number of animals showing the phenotype: 1 (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: -1.0% (includes death as larva, prepupa, pupa)
Lethalities 22 days after larval injection: -1.0% (includes death as larva, prepupa, pupa, adult)
- >TC015351 RA
MILSPDMSL KTYKKRSGT SDQGKEYEW LLCVYYAIQ LSLSSEISD FELCTNNAF FGDFDDVTL RVRFKDGVV KVYLLQTKH SEQPKTVTV KALTSDEGG LTIYKYEKS YENTLQLSN FVYILFTNR TAKFKDKTQ LDEKISLRH SQCVARNFL NTAKNSVYK FESEQNSDF YQRFYLYTN QKNVSNLHK SIADLLQTH FQTDIKSTF VDFMRMWWA RNFMLSKSD IVAKLVELT LSPMIQTLT CDIDNEKTK LLQAAIMRF DLTLVEQNG GSLITQIWH PTLDNHTIK TVSMTGLKY GLVTQGVSN LKQLTRDQQ SRIAWYMNL WPLVVKVDE TKRNQLNCA LNLLKQNKK RVVLLDHDV DMNGFHIFR NLSDLLEGD TLRSAIMQK FNCSLQGKL DVTLAELGS CLMSHFKVD QLVQIATGT NFNIGEPLE ILPPTYIER KFPKIILDC KVIEDSKQW KICLSCNNQ KEQFREKIT KRMVKITTA VTGFANFPK HYVYATDGE YPKELTEEK YRYYKADCH FRVTGQNVQ WLSNRDRCY EFINEIVKY VKNSNEICT ERQLDDNFH CNVKVLTSH TGSGKTSLL KSVKNRALT NCFVMLFDL REHSTFFQK SQTSKEVWN YVTKFYAKN RKYNEFETG LLKEFPKQQ TLILWDSFD ELSFGSREK FVKVVSELR MLGVSQWIT SKNHFRQSL ENEFQVFAL SMVPFHQID QIQYLEKRC GVSVNLIES NFLSNCQIL DNQEYLGIP LQLYIISDI FNKNPKLLE GVYSLADMF RALIEGRYQ HFLSKHPNH EFLTHIIND SREYRLDQY KIAAVKTHL SHVYDKLNL KNTARFIED VQNGDFLGL VNCQDDEFV FEFKILGEY LAALFLSEN CQMLDLEFI FEVKYRNIR FLFDLIMAE GQPDLISLL YKNFDRFDN FDTRDKMGR NALHLICSY GIKHPLLRT EDEKYAFSQ GDLVHEDAP LVKQLVKNL TLKCDVGEK DKLFNWTCF DYADRSLSL GLLELLPNL DNVLPTLSN FNDMSTVLY YALKFDYQN LFKSAQRDI TFVKDTNGV NFLHVATEF GREHFLTWL LTQKVYIQG INMQNSSHW APIQIASFN GNLTFLKLL QTKGATFPL RDPSLVSLA TSHPHRHIL DFLIECECS VNEFYHHNY FTRSLPLAI SHGLVDIVK FLVKNGAKF YNVGKNNMN ALHMALEHQ QYEIADVFL EHGINIDQV DGKQQSSVH HAAQNNHPD VIEWLLARG AKINIFDKN QRNALHWAA HSGSLEAAK VLIKAGIDI DVSDDQGLT PLHLAVLSG QLSIVTLLL DAKCDFKPV NVYKRTPLH LAAMNDLTD IVQVLLQQS NSVDCVESL YGRTPLHYA AWNGHPDCV KTLLLSNAS FDIRCNFGY TPLHLATEA ENYDCCRLL LEAGARFDI ANRNGTTAL DLVNSLDIF MLMMTFDTY PIHTATSKG NYVAIPEIV TLGHDINQF DSQGMRPLH LAVNLYQPF NTVRTLLDN GANVNLRDK DGYKPLHHL LQGRCPDRD TFDLLVTGR GELEINAKT VKNLTPLHI AAQAGCFIL VPGEDTIFE DLTQLLLDK GANIEACDD DGFTPLHYA AQSGNLTIV KLLTRDNLI NQINRYIRT PLHMAAVKG HHFVVKYLI DRGAQQIND YEGDTPLDD ALKIESIET IKLLTNDLS VTNRKKRNV LHMAAKQGL LDLVETFQK DGIMTSEDC DGTSPAEEA LYNRHFKVF DIMDLSNFD VNKYSRRYK KYPLHVATN FQKLRLVEK FVEMGARTD VCDSDGNTP FYNCDDFYI LQLLVKSDN SGLNVQNNK GETVLHFLA KLKHILLEF LVDQGGDVN LMDNDGNTV LDILLNRGI IVKNVLFKT NGFDWNKTN RNGDTLLHI AINYNMADV VQWLLDRGA STGIKNLVI YALRNSHAE IVRLLIKYC DLSQEEDLI YYTVRYGWK LLPELRPHI SPSALTPSE QTDLLESVC HVDYFMDVW DLLPDADVS TMATDLGDT PLHLAVSRG NLTLTKRLL KSGLKPNDA NRSGETPTQ CAVKRGHFS ILYLLLEND KNAVIRSDQ KGNTLLHDA ASEGNLEIV KFLLKKGFR NDICNGRMC TAADVAKEK GHEEVYQFL KNL
- >TC015351 RA
atgattctt tcgccagat atgtcattg aagacctat aaaaaaaga agcggcact tccgaccaa ggcaaggaa tacgaatgg ctgctctgc gtttattac gctatacaa ttatccttg agttcggaa atttccgac tttgagctg tgtacaaat aatgcattt ttcggtgat ttcgatgat gtcacctta agagttcgc ttcaaagat ggcgtcgtt aaggtttac cttctccaa acaaaacac agcgaacaa ccaaaaacc gtcactgtt aaagctctg acgtcagac gaaggcggt ttgacaatc tacaaatac gaaaaatcc tacgaaaat accctacaa ctgtcaaac tttgtctac attttgttt acaaatcgg acagccaaa ttcaaggac aaaacacag cttgatgag aagattagt ttaagacat tcgcaatgt gttgctcga aactttctc aacactgcc aaaaattca gtctataaa tttgaaagc gagcaaaat tccgacttt taccaaagg ttttatctc tataccaat caaaaaaac gtttcgaat ttacacaaa tcaatcgcc gatttactc caaactcac tttcaaacc gacatcaaa tcaactttc gtcgatttc atgcgaatg tggtgggcg agaaatttc atgttgtca aaatcagac attgttgcc aaattggtt gaattgaca ctatcacca atgatacaa acgctaacg tgtgacatt gacaacgaa aaaaccaaa ctactacaa gccgccatt atgcgattt gatttgaca ttagtcgaa caaaatggc ggttcgttg ataacacaa atatggcat ccgacactt gacaaccat acaatcaaa actgtcagt atgacaggt ctcaagtac ggtctagtt acccaaggt gtgtcaaat ttgaagcaa ttgacacgt gaccaacaa tcacgcatc gcttggtac atgaatttg tggcctttg gtcgtaaaa gtcgacgaa accaaacga aatcaactc aactgtgct ttaaacctt ttaaaacaa aacaaaaaa cgtgttgtt ttattggac catgatgtt gatatgaat ggttttcac attttccgg aacttgtca gatttgctt gaaggtgac actttacgt tctgcgatt atgcaaaaa ttcaactgt tcgttacaa ggcaaactt gatgtgaca ctagccgaa cttgggtcg tgtctaatg tcacacttc aaggttgac cagcttgtc caaattgcc actggcacc aactttaac attggagag ccactagaa atattgccg ccaacttac atcgagcga aaattcccc aaaattatc ctcgactgt aaagtgatt gaggattcc aaacagtgg aaaatctgt ctcagttgt aacaaccaa aaggagcaa tttcgcgaa aaaattact aaacgtatg gttaagatc acaacggct gtgactggt tttgctaat tttcccaag cattatgtc tatgctact gatggggaa taccctaag gaactaaca gaggaaaaa tatcgttat tacaaggcc gattgtcac ttccgtgtc actggacag aacgtacag tggttgtca aatcgtgat aggtgttat gaatttatc aacgaaatt gtcaaatat gtcaagaat tcgaatgaa atttgcacc gagcgacaa cttgacgat aatttccat tgtaatgtc aaagttttg acatcgcat acgggcagt ggcaaaacc agtcttttg aaaagtgtc aaaaaccga gctttgact aattgcttt gtcatgttg tttgacttg agggaacat tcaacattt ttccaaaag tctcagact tcgaaggaa gtttggaat tatgtgaca aaattttac gccaaaaat cgtaaatat aacgaattt gagactgga cttctcaag gaatttcca aaacaacaa actttaata ctttgggat agtttcgac gaattgtca ttcgggtca agggaaaaa tttgtcaaa gttgtgtca gaacttcgc atgttggga gtttctcaa tggataact tcgaaaaat cattttcgt caatctctg gaaaacgaa tttcaagtt ttcgcatta tctatggta ccgttccac caaatcgac caaatccaa tacctggag aagcgctgt ggtgtcagt gttaattta atcgagtca aatttcttg tcaaattgt caaatctta gacaatcag gagtacctg ggcattccc ctccaactc tacatcatt agcgacatc ttcaataaa aaccctaaa ctactcgaa ggggtttac agtcttgcc gacatgttt cgtgcatta attgaggga cgttaccaa catttttta tcaaaacac cccaatcat gaatttttg acacacatt attaatgat agtcgagag taccgtttg gaccaatac aaaatagct gctgtcaag acacaccta agtcatgtt tatgacaaa ctcaacttg aaaaatacc gcacgtttt atagaagac gtacaaaat ggcgatttt ttgggactt gtcaattgt caagatgac gagtttgtt tttgagttt aaaatactt ggggaatat cttgctgca ttgtttctt tcggaaaat tgtcaaatg ttggatctt gaatttatt tttgaagtc aaatatcga aatattcgc tttttattc gatttgatt atggctgag ggtcagcca gatttaatt tcactactt tataaaaac tttgacagg tttgacaat tttgacacg agggacaaa atgggtcga aatgcccta cacttgatt tgttcttac ggcataaag cacccactt ttacgcacg gaagacgag aaatatgcc ttcagtcaa ggggatctg gtacatgaa gatgcccca cttgtcaaa caacttgtc aaaaatttg acactcaag tgtgacgtt ggggaaaaa gacaaattg ttcaactgg acgtgtttt gattacgcc gatcgttca ctctcactt ggccttctc gaacttttg ccaaatctt gacaacgtt cttccaaca ttgtcaaat ttcaatgac atgtccacc gtactgtat tatgccctt aaatttgac taccaaaac ttattcaaa tctgcacaa agagatata acgtttgtg aaagacacc aatggtgtt aatttcctt catgttgca acagaattc ggtcgtgag cattttttg acatggctt ttgacacaa aaagtctac atccaaggc attaatatg caaaattcg agtcattgg gcaccgatt caaattgcc tcctttaat ggcaatttg acatttttg aagcttttg caaaccaaa ggtgcaacg tttccatta cgtgatccc tcacttgtg tcactcgct acgtcgcac cctcatcgt cacattctc gactttttg atagaatgt gaatgtagc gtgaatgaa ttctaccat cacaattac ttcacacga tcgttacct ctggcgata agtcatggg ctagttgac attgttaaa tttttagtc aaaaacggg gcaaaattc tacaacgtt ggcaaaaac aacatgaat gccttgcat atggcttta gaacatcaa caatatgaa attgcggac gtgttcctt gaacatggc attaatatt gatcaagtt gacggcaag caacaatca agtgttcat catgccgca caaaacaac caccctgat gtaatcgag tggttgttg gcacgtggg gcaaaaatt aatattttt gacaagaat caacgcaat gctttgcat tgggcggca cattccggg tcacttgaa gccgcaaaa gtgctgatc aaagctgga attgacatt gatgtttct gatgatcaa ggtttgaca ccgttacat ttagcagta ttgtcagga caattgtca attgttacg ttactactg gacgcaaag tgtgacttt aaaccagtc aatgtgtac aaaaggaca ccgttacat cttgccgca atgaacgat ttgactgac attgtccaa gttttacta caacagtca aattcagtt gactgtgtt gaaagtctc tacggacga acaccgtta cattatgcc gcttggaac ggtcatcct gactgtgtc aaaacccta ctattgtca aatgccagt tttgacatt cgatgcaat ttcggttac acaccgtta catcttgcc acagaagcg gaaaattac gattgttgt cgtttgtta ctcgaagct ggtgctaga tttgacatt gctaaccgt aacggtact acagcactc gatctagtt aacagtctt gacattttc atgctcatg atgacattt gacacatat ccaatacat acagcaacg tctaaaggc aattacgtt gcaatccct gagattgta acacttgga catgacatc aaccaattt gacagtcaa gggatgaga ccgttacac ctagctgtc aatttgtac caaccgttc aacactgtc agaactcta cttgacaat ggagccaat gtcaatttg cgagacaaa gatgggtat aaacctctc catcacttg ttacaaggt cgttgtccc gatcgagac acttttgac ttacttgta acgggtcga ggtgagctt gaaatcaac gcaaaaact gtcaaaaac ttgacaccg ttacatatc gcagcacag gccggttgt tttatacta gtaccaggt gaagacaca atttttgaa gatttgaca cagctattg cttgacaaa ggggcaaat attgaagca tgtgatgat gatggtttt acaccgtta cattacgca gcacagagt ggtaatttg acaattgtc aaacttttg acacgtgac aatttaatt aatcaaatc aatcgatac attcggaca ccgttacat atggctgct gtcaaaggt caccacttt gttgtcaaa tatttaata gatagaggg gcacaacaa atcaatgac tacgaaggt gatactcca ctagatgat gctttaaaa attgaaagt attgaaact atcaaatta ttaacgaat gatttgtct gtgaccaat cgtaaaaaa cggaacgtt ttacatatg gctgcgaaa caaggcctg cttgattta gtcgaaaca tttcaaaaa gatggaatt atgactagt gaggattgt gatggaaca tcaccagct gaggaagct ttatataac cgccatttt aaggttttt gacattatg gacttgtca aattttgac gttaataaa tacagtcga cggtataag aaatatcca cttcatgtg gccactaat tttcaaaaa ctaagacta gttgagaag tttgtcgaa atgggtgcc agaactgac gtgtgtgac agtgatggc aatacgccg ttttataat tgtgatgat ttctacatt ttgcaattg ttggtcaag tctgacaat agtggtcta aatgtgcaa aataacaag ggagagacc gttttacat tttttggca aaattgaaa cacatttta ttggaattt ttggtggat caaggtggc gatgtcaat ttaatggac aatgacggt aacactgtt ttggatata ttgttgaat cgcgggatt atcgtgaaa aatgtgttg tttaaaacg aatggtttt gactggaac aaaacaaat cgtaatggt gatacttta ctccatatt gctatcaat tacaatatg gcggatgtc gtgcaatgg ctacttgat cgaggtgcc tcaactgga ataaaaaat cttgtgatt tacgcttta cgaaattct catgctgaa attgttcga ttattaata aaatattgt gatttgagt caggaggag gatcttatt tactatact gtacgatac ggatggaaa ctcctccca gaactccgc ccacatatc tccccctcc gctctcaca ccaagcgag caaaccgat ctcctcgaa tcagtttgt cacgttgat tatttcatg gacgtgtgg gatttactc cccgacgct gatgtctcc accatggca accgatctc ggtgacaca cctctacat ttggctgtt tctcgtggc aacttgact ttgactaaa cgtttactt aaaagcggt ttaaaaccg aatgatgct aatcgtagt ggtgaaaca ccgacacaa tgtgcagtc aagagagga catttctca attttatac cttttactc gagaatgat aaaaatgca gtgattcga agcgaccag aaaggaaat actttacta catgatgca gcaagtgag gggaatttg gaaattgtc aagtttcta ctaaagaag ggctttcgg aacgatatt tgcaatggg agaatgtgt actgcagcc gatgtagca aaggagaag ggacatgaa gaggtttac cagtttttg aaaaatttg taa
- >TC015351 RA
tagacagtt ggttttttc actgagcac attttcttg ttgaaatca cgcaaaaaa atgaatgaa tgtttgtgt agtgcattt ttgtgatga ttctttcgc cagatatgt cattgaaga cctataaaa aaagaagcg gcacttccg accaaggca aggaatacg aatggctgc tctgcgttt attacgcta tacaattat ccttgagtt cggaaattt ccgactttg agctgtgta caaataatg catttttcg gtgatttcg atgatgtca ccttaagag ttcgcttca aagatggcg tcgttaagg tttaccttc tccaaacaa aacacagcg aacaaccaa aaaccgtca ctgttaaag ctctgacgt cagacgaag gcggtttga caatctaca aatacgaaa aatcctacg aaaataccc tacaactgt caaactttg tctacattt tgtttacaa atcggacag ccaaattca aggacaaaa cacagcttg atgagaaga ttagtttaa gacattcgc aatgtgttg ctcgaaact ttctcaaca ctgccaaaa attcagtct ataaatttg aaagcgagc aaaattccg acttttacc aaaggtttt atctctata ccaatcaaa aaaacgttt cgaatttac acaaatcaa tcgccgatt tactccaaa ctcactttc aaaccgaca tcaaatcaa ctttcgtcg atttcatgc gaatgtggt gggcgagaa atttcatgt tgtcaaaat cagacattg ttgccaaat tggttgaat tgacactat caccaatga tacaaacgc taacgtgtg acattgaca acgaaaaaa ccaaactac tacaagccg ccattatgc gatttgatt tgacattag tcgaacaaa atggcggtt cgttgataa cacaaatat ggcatccga cacttgaca accatacaa tcaaaactg tcagtatga caggtctca agtacggtc tagttaccc aaggtgtgt caaatttga agcaattga cacgtgacc aacaatcac gcatcgctt ggtacatga atttgtggc ctttggtcg taaaagtcg acgaaacca aacgaaatc aactcaact gtgctttaa accttttaa aacaaaaca aaaaacgtg ttgttttat tggaccatg atgttgata tgaatggtt ttcacattt tccggaact tgtcagatt tgcttgaag gtgacactt tacgttctg cgattatgc aaaaattca actgttcgt tacaaggca aacttgatg tgacactag ccgaacttg ggtcgtgtc taatgtcac acttcaagg ttgaccagc ttgtccaaa ttgccactg gcaccaact ttaacattg gagagccac tagaaatat tgccgccaa cttacatcg agcgaaaat tccccaaaa ttatcctcg actgtaaag tgattgagg attccaaac agtggaaaa tctgtctca gttgtaaca accaaaagg agcaatttc gcgaaaaaa ttactaaac gtatggtta agatcacaa cggctgtga ctggttttg ctaattttc ccaagcatt atgtctatg ctactgatg gggaatacc ctaaggaac taacagagg aaaaatatc gttattaca aggccgatt gtcacttcc gtgtcactg gacagaacg tacagtggt tgtcaaatc gtgataggt gttatgaat ttatcaacg aaattgtca aatatgtca agaattcga atgaaattt gcaccgagc gacaacttg acgataatt tccattgta atgtcaaag ttttgacat cgcatacgg gcagtggca aaaccagtc ttttgaaaa gtgtcaaaa accgagctt tgactaatt gctttgtca tgttgtttg acttgaggg aacattcaa catttttcc aaaagtctc agacttcga aggaagttt ggaattatg tgacaaaat tttacgcca aaaatcgta aatataacg aatttgaga ctggacttc tcaaggaat ttccaaaac aacaaactt taatacttt gggatagtt tcgacgaat tgtcattcg ggtcaaggg aaaaatttg tcaaagttg tgtcagaac ttcgcatgt tgggagttt ctcaatgga taacttcga aaaatcatt ttcgtcaat ctctggaaa acgaatttc aagttttcg cattatcta tggtaccgt tccaccaaa tcgaccaaa tccaatacc tggagaagc gctgtggtg tcagtgtta atttaatcg agtcaaatt tcttgtcaa attgtcaaa tcttagaca atcaggagt acctgggca ttcccctcc aactctaca tcattagcg acatcttca ataaaaacc ctaaactac tcgaagggg tttacagtc ttgccgaca tgtttcgtg cattaattg agggacgtt accaacatt ttttatcaa aacacccca atcatgaat ttttgacac acattatta atgatagtc gagagtacc gtttggacc aatacaaaa tagctgctg tcaagacac acctaagtc atgtttatg acaaactca acttgaaaa ataccgcac gttttatag aagacgtac aaaatggcg attttttgg gacttgtca attgtcaag atgacgagt ttgtttttg agtttaaaa tacttgggg aatatcttg ctgcattgt ttctttcgg aaaattgtc aaatgttgg atcttgaat ttatttttg aagtcaaat atcgaaata ttcgctttt tattcgatt tgattatgg ctgagggtc agccagatt taatttcac tactttata aaaactttg acaggtttg acaattttg acacgaggg acaaaatgg gtcgaaatg ccctacact tgatttgtt cttacggca taaagcacc cacttttac gcacggaag acgagaaat atgccttca gtcaagggg atctggtac atgaagatg ccccacttg tcaaacaac ttgtcaaaa atttgacac tcaagtgtg acgttgggg aaaaagaca aattgttca actggacgt gttttgatt acgccgatc gttcactct cacttggcc ttctcgaac ttttgccaa atcttgaca acgttcttc caacattgt caaatttca atgacatgt ccaccgtac tgtattatg cccttaaat ttgactacc aaaacttat tcaaatctg cacaaagag atataacgt ttgtgaaag acaccaatg gtgttaatt tccttcatg ttgcaacag aattcggtc gtgagcatt ttttgacat ggcttttga cacaaaaag tctacatcc aaggcatta atatgcaaa attcgagtc attgggcac cgattcaaa ttgcctcct ttaatggca atttgacat ttttgaagc ttttgcaaa ccaaaggtg caacgtttc cattacgtg atccctcac ttgtgtcac tcgctacgt cgcaccctc atcgtcaca ttctcgact ttttgatag aatgtgaat gtagcgtga atgaattct accatcaca attacttca cacgatcgt tacctctgg cgataagtc atgggctag ttgacattg ttaaatttt tagtcaaaa acggggcaa aattctaca acgttggca aaaacaaca tgaatgcct tgcatatgg ctttagaac atcaacaat atgaaattg cggacgtgt tccttgaac atggcatta atattgatc aagttgacg gcaagcaac aatcaagtg ttcatcatg ccgcacaaa acaaccacc ctgatgtaa tcgagtggt tgttggcac gtggggcaa aaattaata tttttgaca agaatcaac gcaatgctt tgcattggg cggcacatt ccgggtcac ttgaagccg caaaagtgc tgatcaaag ctggaattg acattgatg tttctgatg atcaaggtt tgacaccgt tacatttag cagtattgt caggacaat tgtcaattg ttacgttac tactggacg caaagtgtg actttaaac cagtcaatg tgtacaaaa ggacaccgt tacatcttg ccgcaatga acgatttga ctgacattg tccaagttt tactacaac agtcaaatt cagttgact gtgttgaaa gtctctacg gacgaacac cgttacatt atgccgctt ggaacggtc atcctgact gtgtcaaaa ccctactat tgtcaaatg ccagttttg acattcgat gcaatttcg gttacacac cgttacatc ttgccacag aagcggaaa attacgatt gttgtcgtt tgttactcg aagctggtg ctagatttg acattgcta accgtaacg gtactacag cactcgatc tagttaaca gtcttgaca ttttcatgc tcatgatga catttgaca catatccaa tacatacag caacgtcta aaggcaatt acgttgcaa tccctgaga ttgtaacac ttggacatg acatcaacc aatttgaca gtcaaggga tgagaccgt tacacctag ctgtcaatt tgtaccaac cgttcaaca ctgtcagaa ctctacttg acaatggag ccaatgtca atttgcgag acaaagatg ggtataaac ctctccatc acttgttac aaggtcgtt gtcccgatc gagacactt ttgacttac ttgtaacgg gtcgaggtg agcttgaaa tcaacgcaa aaactgtca aaaacttga caccgttac atatcgcag cacaggccg gttgtttta tactagtac caggtgaag acacaattt ttgaagatt tgacacagc tattgcttg acaaagggg caaatattg aagcatgtg atgatgatg gttttacac cgttacatt acgcagcac agagtggta atttgacaa ttgtcaaac ttttgacac gtgacaatt taattaatc aaatcaatc gatacattc ggacaccgt tacatatgg ctgctgtca aaggtcacc actttgttg tcaaatatt taatagata gaggggcac aacaaatca atgactacg aaggtgata ctccactag atgatgctt taaaaattg aaagtattg aaactatca aattattaa cgaatgatt tgtctgtga ccaatcgta aaaaacgga acgttttac atatggctg cgaaacaag gcctgcttg atttagtcg aaacatttc aaaaagatg gaattatga ctagtgagg attgtgatg gaacatcac cagctgagg aagctttat ataaccgcc attttaagg tttttgaca ttatggact tgtcaaatt ttgacgtta ataaataca gtcgacggt ataagaaat atccacttc atgtggcca ctaattttc aaaaactaa gactagttg agaagtttg tcgaaatgg gtgccagaa ctgacgtgt gtgacagtg atggcaata cgccgtttt ataattgtg atgatttct acattttgc aattgttgg tcaagtctg acaatagtg gtctaaatg tgcaaaata acaagggag agaccgttt tacattttt tggcaaaat tgaaacaca ttttattgg aatttttgg tggatcaag gtggcgatg tcaatttaa tggacaatg acggtaaca ctgttttgg atatattgt tgaatcgcg ggattatcg tgaaaaatg tgttgttta aaacgaatg gttttgact ggaacaaaa caaatcgta atggtgata ctttactcc atattgcta tcaattaca atatggcgg atgtcgtgc aatggctac ttgatcgag gtgcctcaa ctggaataa aaaatcttg tgatttacg ctttacgaa attctcatg ctgaaattg ttcgattat taataaaat attgtgatt tgagtcagg aggaggatc ttatttact atactgtac gatacggat ggaaactcc tcccagaac tccgcccac atatctccc cctccgctc tcacaccaa gcgagcaaa ccgatctcc tcgaatcag tttgtcacg ttgattatt tcatggacg tgtgggatt tactccccg acgctgatg tctccacca tggcaaccg atctcggtg acacacctc tacatttgg ctgtttctc gtggcaact tgactttga ctaaacgtt tacttaaaa gcggtttaa aaccgaatg atgctaatc gtagtggtg aaacaccga cacaatgtg cagtcaaga gaggacatt tctcaattt tataccttt tactcgaga atgataaaa atgcagtga ttcgaagcg accagaaag gaaatactt tactacatg atgcagcaa gtgagggga atttggaaa ttgtcaagt ttctactaa agaagggct ttcggaacg atatttgca atgggagaa tgtgtactg cagccgatg tagcaaagg agaagggac atgaagagg tttaccagt ttttgaaaa atttgtaat aaagtttct gggggaaaa accgtgttt tatttgttt ga
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