| General information | |||
| TC005546 | Gene
The sequence from the iB fragment is matched against the genes from the official gene set. The matche(s) is/are visualizied in a genome browser. |
NC_007423.3: 11452895..11462117 | |
| 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 |
|
TC005546 | |
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_00881 | 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: 10.0% (includes death as pupa, adult)
eclosion not fulfilled (with pupal and adult features) - Number of animals showing the phenotype: 1 (3 dpi)Days post injection
Fertility
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: 100.0% (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 not fulfilled (died as pupa) - Number of animals showing the phenotype: 9 (11 dpi)
Days post injection
- >TC005546 RA
MSTLAMQQF VAGLKSRNP DVRQKTARE LSLYVKSEL REATPDEIT NFLDEFNHH IFEMVSSND PNEKKGGIL AIVCLIGVD FGNMTTRIS RFANYLRNL LPSSDVSVM ESVAKTMGR LALVSGSKA SEYVEFEVK RAFEWLSGD RIEGKRQAA VLVLKELAL TMPTYFYQH VSQFFDLIF FAIQDPKPA IRESAVEAL RAALVVTAQ RETAKQNQK PQWYKQCYD ESVKMLSVE RGERMKEER VHGFLLILN ELVRCSNAE WERKCKSLL ERTDTKQPV ETSFNFTKS RFVFSVSRR IPQYQAVSL APSQIAIIE SQSCRGLVS EKFDYICLD VLAQRLSRS SHIQQSFLT ILPRLAAFD RENYVKQHL SPVMNHIFN ILKSREKER AKAFVTIGL IAIAVESDI EPYIERIME IIRMYLPRA DILGRKRSP IDSSIFNCV TFLAHALKN HSKMDIPNL LEPMLATGL TASLTICFR ELSRKVPEH KERISLGLL KMLSYILLN KPLVHPGMP RHLTGTVMS LAIPEVNDV QIIVLALKT LGTFDFEGQ RLLPFVHRC ANHFLIHDN NEIRLEAVR TTCRLLRHA IHSTAKNSS DTVTKTVAA VLHRLLSVG LTDTDPNVR YGVLISLDR TFDNHLAQA ESLSALFLA LQDEMFEIR EVALFTIGR LSAMNPAYV MPSLRKTLV QFLSELEHS GSGRNKEQG ARMLDHLVV SAPRMIRPY MEPILKVLV PKLRDPEPN PGVVLSVLL TIGDLAEVS SGGTELQEW MQELMGILL EMLGDASAP EKRGAALCT LGQLVGVTG HVIQPYTEY PILLDVLLN FLKTEQQSY IRRETIRVL GLLGALDPY KHKINRGEI DYYPEAPVL IPMTDKGED LNADLTSSE MLVNMSSST LEEYYLAIA IATLMRIIK DPTLAQHHT MVVQAVTFI FKSLGIKCV PYISQVLPS LLHVVRTAD VNFKEFLFQ QLAQLIYIV KQHIRNYLD DICLLIKEF WTPNSTIQG TLILLVEHI AVALGAQFK VYLPKMLPH ILRVLNHDT SKERLYTIK LLEALHNFG NNLDEYMHL ILPPIVRLF DAQECPIVV SKKALETID QLAEIIDFS DFISRIVHP LVRTIDNCP ELRPTAMET LCSLMQQLG RKFSIFVPL VQKVMTKHK IQHSKFDTL VSKIQYETT LAEDVDFPM PRSKTTGKN RDPAMPADS GMIQRLKVS ESNLQQAWT PVRRVSKDD WLEWLRRLS IELLKQSPI PALRSCLTL AQTYSQLPR DLFNAAFVS CWSELSENM QNELISCLE QALTVPDVP EITQTILNL AEFMEHCDK GPLPLDSHI LGHHAMHCR AYAKALHYK EEEFQRGAS SQVVEALIS INNKLQQKE AAEGLLQYV MQREMQVQV RWYEKLHNW EKALRLYTE KLEGDEADQ EACLGKMRC LEALGEWGE LHQVVEKSF SLLNDDSKQ KASRLAAAA SFGLHDYRS METYVNVIP RDTQEGAFY RAILAIHKE DYEVAQRFI DSARDLLDN ELTAMAGES YQRAYGAMV MVQMLSELE EVMQYRLVP ERRPTLKAM WWQRLQSGQ KLVEDWQRI IQVHSLVLT PQEDKRTWV KYASLCRKS GSLMLSQKT LVMLLGYDP SERPDAPLP KNQPHITLA YAKHLWVAQ EKQKAFQKL SQFVADYSQ NEPNDDVTV EEKKRLLAR CYLKLGAWH EALEGINET SIPFILKCY KQATEHDPQ WYKAWHAWA YMNFETVLY YTRQEDKTH YTVLAVQGF FKSINLSKG SSLQDTLRL LTLWFDYGD WPEVYDAIV EGIRLVEKN TWLQVIPQL IARIDTTAL VSKLINHLL VDIGKTHPQ ALVYPLTVA TKSNSIRRR NAANSILKS MSEHSPTLV SQAMMASEE LIRVAILWH EIWHEGLEE ASRLYFGER NVKGMLRIL EPLHAMMER GPQTLKETS FNQTYGRDL NEAQDWCQR YKLSSNIRD LNQAWDLYY HVFRRISRQ LPQLTSLEL QYVSPNLLV CQDLELAVP GSYCPGQPI VRIANFNRS LEVITSKQR PRKLVIRGS NGKDYMFLL KGHEDLRQD ERVMQLFGL VNTLLMKDP DTFRRNLTI QRYAVIPLS TNSGLIGWL PHCDTLHTL IRDYRDKKK ILLNIEHRI MLRMAPDYD HLTVMQKME VFEHALEHT HGDDLARLL WLKSPSSEV WFDRRTNYT RSLAVMSMV GYILGLGDR HPSNLMLDR LSGKILHID FGDCFEVAM TREKFPEKI PFRLTRMLI NAMEVTGIE GTYRRTCES VMSMLHRNK DSLMAVLEA FVYDPLLNW RLMDIEVRN LPMNSDSMS TSCSQKQGS SESLNVPKR EMIMEDNEP QHEVNARAV IIINRVRDK LTGNDFATE EPLTIPKQV DLLIQQATS NENLSQCYI GWCPFW
- >TC005546 RA
atgtccact ttggccatg caacaattc gttgcgggg ttgaaatcc cgcaacccc gacgttcgt caaaaaacc gcacgagaa ttaagcttg tacgtaaaa agcgagctt cgcgaagcc acccccgac gaaataaca aacttcctt gacgaattc aaccaccac atcttcgaa atggtctcg agcaacgac cccaacgag aaaaaaggc ggaattttg gcgattgta tgtttgatt ggagtcgat ttcgggaac atgaccaca cgtatatca cgcttcgcc aattattta cgcaacttg ttgccctcg agcgacgtg agcgtcatg gagtcggtg gcgaagaca atggggcgc ttggcgctg gtttcaggc tcgaaagcc tccgaatac gtcgagttt gaagttaag agggccttt gagtggctc agtggcgac cggattgaa gggaagcga caagctgcc gttttagtc ttgaaagag ttggcacta actatgccg acgtatttc taccaacat gtgtcgcaa tttttcgat ctgattttt ttcgcgata caggacccc aaacctgcc attagggag agcgctgtg gaggctttg agggccgct ttggtcgtc acagcgcaa cgagagacc gccaaacag aaccaaaag ccacaatgg tacaaacaa tgttacgac gaatccgtg aaaatgctc tcggtggaa agaggcgaa cgcatgaaa gaggagcgc gtccacggc tttctcctc attctcaac gaactggtg cgttgctca aacgccgaa tgggaacgc aaatgcaaa agtttgctc gaacgcact gacaccaaa caaccggtc gagaccagc ttcaatttt actaaatct cgcttcgtt ttttcggtc tctcgacgc atcccccaa taccaagct gtgtctttg gcaccctct caaatcgca atcatagaa tcccaaagt tgccgaggc ctcgtttcc gagaaattc gactacatt tgtttagac gtcctggct cagagactc tcccgttcg tctcacatc caacaaagt tttctcacg attttaccc cgattggct gcttttgac cgcgaaaac tacgtcaag cagcactta agccccgtc atgaaccac atcttcaac attttgaag agtcgggaa aaggagaga gctaaggct tttgtgact attggctta attgcgatc gctgtcgag agcgacatt gagccgtac attgaacgt attatggaa ataattcgc atgtatttg ccccgagcc gatattttg gggcgcaag cggagcccc atcgactcc tcaattttc aactgtgtc actttccta gcccacgcc ctcaaaaac cactcaaaa atggacatc ccgaacctc ctggaacca atgctagca acgggatta accgcatcc ctcacgata tgtttccgg gaattatcc cgaaaagtc ccggaacat aaagaacga atcagtctc gggttatta aaaatgctg agttatatt ctcctgaat aaaccacta gttcatccc gggatgccc cgccacttg accggaacc gtaatgagt cttgcaata ccagaagtc aacgatgtg caaatcata gttttggcc ctcaaaaca ctcggaacg tttgacttt gagggccag agactcctc ccttttgtc caccgctgc gccaatcat ttcctcatc catgacaat aacgaaatc cgattggag gctgtcaga accacgtgt cggctcctc cgacatgct attcactcc acagcgaaa aactcgtcg gatacggtc accaaaacc gtagctgct gttttacat cgtttgtta agtgttggt ttgaccgat accgaccca aatgtccgg tacggagtc ttgatttcg ctcgatcgt acgtttgat aatcattta gcacaagct gagtcgctc agtgcgctg tttctggcg ttgcaagac gaaatgttt gaaatacgc gaagttgca ctgtttaca atcgggcgc ttgagtgcg atgaatccg gcatatgtc atgccctcg ttgaggaaa actttagtg cagttttta tccgaattg gaacattcg ggcagtggg aggaataag gaacaaggg gcgaggatg ttagatcat ttggttgtg agcgccccg aggatgata aggccctac atggagccc attttgaag gtcctggtg cccaaactg cgcgacccc gagccaaac cccggcgtc gtgctttcc gtcctgctc acaatcggc gatttggcc gaggtgagc agcggaggc accgaactg caagaatgg atgcaagaa ctgatgggt attttactc gaaatgttg ggcgacgct tccgccccc gaaaaacgg ggcgccgcc ttatgcacc ttgggccag ttggttgga gtgaccggt cacgtaatc caaccctac accgaatat ccaatactg ctcgacgtc ctcctcaat ttcctcaaa accgaacaa cagtcctac attcggcga gaaacaatc cgcgttttg gggctgttg ggggctttg gatccgtac aaacataaa ataaatcgg ggggagatt gattattat cccgaagcg cccgttttg ataccgatg acggataaa ggggaggat ttgaacgcg gatttgacg tcgagcgaa atgctggtt aatatgagt tcgagcact ttggaggag tactatttg gcgatagct atagccact ttgatgcgg attattaag gatccgacg ctggcacaa caccataca atggtggtg caagcagtt actttcatt tttaaaagt ttggggatt aagtgtgtg ccgtatata tcgcaagtt ttgcccagt cttttgcac gtagtacgg acagccgac gttaacttt aaggaattc ctcttccaa cagttggca caactaatc tacatcgtc aagcaacac attcgtaac tacttggac gacatttgc ctactaatc aaagaattt tggaccccg aacagtacg attcaaggg actttgatc cttttagtt gaacacatc gcagtggcg ttgggggcg caatttaag gtttattta ccgaaaatg ttgccccat atcttacgc gtgttgaac cacgacacg tccaaggaa cgactctac actatcaaa ttattggaa gcgttacac aatttcggt aacaatttg gacgaatat atgcacttg attttgccc cccattgtc cggttattc gacgctcag gagtgtcca atagtcgtg tcgaaaaaa gccctcgag actatcgac caattggcc gaaattatc gacttttcg gactttatc tcacgcatt gtccacccc ttggttcga acaattgat aactgtccc gaattgaga cccacagcc atggaaact ttatgttca ctcatgcaa caattgggg cgaaaattc tcaattttc gtacctttg gtacagaaa gtgatgacc aaacataag atacaacac agtaaattc gatacttta gtttcgaaa atccaatac gagacaaca ctggccgaa gacgttgat ttcccaatg ccgagatcc aaaacaacc gggaaaaat cgagaccca gcgatgcct gccgatagt ggaatgatt caaagactc aaagtttcg gaatccaac ctacaacag gcctggacc cctgtccga agagtctcg aaagacgat tggttggaa tggctaaga cgcctcagt atcgaacta ttaaaacag tcaccaatt ccagcatta cgttcgtgc cttacttta gcacagacg tattcgcaa ctgccacgt gatttattc aacgcagcg ttcgtaagt tgttggtcc gaattgagc gaaaacatg cagaatgaa ctgatcagt tgtctcgag caagctttg actgtgccg gacgttccg gaaataact caaactatt ttgaatttg gccgaattt atggaacat tgcgacaag gggcctttg cccctcgat agccacatt ctggggcac catgcgatg cattgccgg gcgtacgcc aaagccttg cattataag gaagaggag tttcaaagg ggggcgagc agtcaggtt gtcgaagca ctcatttcg atcaataat aagttgcaa caaaaggag gcggctgaa gggttgtta caatacgtc atgcagagg gaaatgcaa gtgcaggtc aggtggtac gagaagttg cacaattgg gagaaggcc ctgaggttg tatactgag aaactggaa ggggacgag gcggaccaa gaggcttgt ttgggcaaa atgagatgt ctcgaagcg ttgggcgaa tggggcgaa ctgcaccaa gtcgtcgag aaaagtttc agtttacta aatgacgac agcaaacaa aaggcgagt cggctagcg gctgcggcc tccttcggt ttgcacgat tacaggtca atggaaact tacgttaat gtaattcca agggatacg caagagggt gcgttctac cgtgcaatt ttggcaata cataaagag gattacgaa gtggcccaa agattcatc gattcagct cgcgattta ctcgataat gaattaact gcgatggct ggtgagtcc taccaaagg gcctacggg gccatggtc atggtacaa atgttatcc gagttggaa gaagtaatg cagtatcgg ttagtacca gaacggagg cccacttta aaagccatg tggtggcaa aggctccaa tcgggccaa aaactggtc gaagactgg caaagaatc atccaagtc cattccttg gtcttgacc ccacaagaa gacaaacga acgtgggtg aaatacgcg tcattgtgt cgcaagagc ggttcatta atgctatcg caaaaaacc ctagtcatg ttactcggt tacgaccca agcgaacgt cccgacgcc cccttgccc aaaaaccag cctcacatc acactagcc tacgccaaa catttatgg gtggcacag gaaaaacaa aaagcgttc caaaaacta tcacaattc gtcgctgac tattcgcaa aacgagcca aacgacgac gtgacagtt gaagagaaa aaaagactc ctggctagg tgttacctc aaactgggg gcctggcat gaggcactt gaaggcatc aacgaaacc tccatacct ttcattttg aagtgttac aaacaagcg actgaacac gaccctcaa tggtacaag gcatggcac gcttgggcc tatatgaat ttcgaaacg gtcttgtat tacacgaga caggaggac aagacgcac tatacagtc ttagccgtt cagggattt ttcaaatcg attaattta tctaaagga agttcattg caagacact ttacgttta ttaacactg tggttcgat tacggcgat tggcccgaa gtttacgat gccatagtc gaaggcatt cgtttagtc gagaaaaac acgtggctt caagtaatt ccgcaatta atcgcgcga attgacaca acggcgtta gtatcgaaa ttaatcaat catctattg gttgatatt ggaaaaacg catccacaa gccctagtc tatccgtta accgtcgca acaaaaagt aattcaatc agacgtaga aatgcagct aatagtatt ttaaaatcg atgagtgaa cattcccct acgttggta agtcaggcc atgatggcc tcagaagaa ttgatccga gtggcaatt ttgtggcac gaaatctgg cacgaggga ctagaagaa gcctcaaga ttgtatttc ggcgaacgt aacgttaaa ggaatgttg cgaattctc gaaccgttg catgcgatg atggaaagg gggccccag acgttgaaa gagacgagt tttaatcag acgtacggc agggatctg aacgaggcg caagactgg tgccaaaga tacaagctt tcgagcaac attagagac ctgaaccag gcgtgggac ctctactac cacgttttc cgccgcatt tcgcgtcaa ttaccgcaa ctcacctcc ctcgaatta caatacgta tcgccaaat ttgctagtt tgtcaagac ttagagttg gcagtgcct ggtagctac tgtccagga cagccaatc gtacgcatt gccaatttt aaccgatct ctcgaagta attacctca aaacagcgg ccccgaaaa ctagtaatt cgtggaagc aacggcaaa gactacatg tttttactt aaaggtcac gaggatctg agacaagac gagcgtgtt atgcaactg ttcggttta gttaatact ttactaatg aaagacccg gatacgttc aggcggaat ttgacgata cagcggtat gcggtgatt cccctgagt acgaattcg ggtctgatt gggtggtta ccgcactgt gatacgctg cacacttta atacgggat tatagagac aagaagaag attttgttg aatattgag cataggatt atgttacgg atggcgccg gattatgat catttaacc gtgatgcag aaaatggaa gtttttgag cacgcactt gagcacaca catggtgac gatttagcg cggttgttg tggctgaaa agcccctcg tctgaggtt tggtttgac cggagaacg aactacacg cggtcttta gctgtgatg tcaatggtt gggtatatt ttggggttg ggggatcgc cacccctcc aatttaatg ttggacaga ctgagcggc aaaatcctg catatagat ttcggcgat tgcttcgaa gtggcaatg actagggaa aaattcccg gaaaagatc cccttccgt ttgactagg atgctaata aacgccatg gaagtgaca gggattgaa ggcacgtac agacggact tgtgagtcg gtcatgtcc atgttacac cgaaataaa gacagtctg atggcggtg cttgaagcc tttgtttat gaccccctg cttaattgg cgattgatg gatattgag gtgcggaac ttgccgatg aacagcgat tcaatgtcg acgagttgt tcccaaaaa cagggctca tccgaatcg ctcaatgtt ccgaaacgc gagatgata atggaagat aacgaaccg caacacgaa gttaatgcc agggcggtt attatcatc aatcgagtt agagataaa ctgaccggg aacgatttc gctacggag gaacctttg actattcct aagcaagtc gatctgtta atacaacaa gcgacaagt aacgagaac ttgtcacag tgttacata ggctggtgc cccttttgg taa
- >TC005546 RA
gatttatca catatttaa gataccaag tttttacat gattgtaat aaagacaaa aaaatactc tttgtgctc gaataaggc cccattgaa attcaaacc ggccgaaaa atcccccaa gtcattcgg caaaaatgg aagcaccat catgagtgt gataacaat ctttcagtg ttgataaac ccgaaaatg cttcctttc taacgtcgc accgtcacc atgtccact ttggccatg caacaattc gttgcgggg ttgaaatcc cgcaacccc gacgttcgt caaaaaacc gcacgagaa ttaagcttg tacgtaaaa agcgagctt cgcgaagcc acccccgac gaaataaca aacttcctt gacgaattc aaccaccac atcttcgaa atggtctcg agcaacgac cccaacgag aaaaaaggc ggaattttg gcgattgta tgtttgatt ggagtcgat ttcgggaac atgaccaca cgtatatca cgcttcgcc aattattta cgcaacttg ttgccctcg agcgacgtg agcgtcatg gagtcggtg gcgaagaca atggggcgc ttggcgctg gtttcaggc tcgaaagcc tccgaatac gtcgagttt gaagttaag agggccttt gagtggctc agtggcgac cggattgaa gggaagcga caagctgcc gttttagtc ttgaaagag ttggcacta actatgccg acgtatttc taccaacat gtgtcgcaa tttttcgat ctgattttt ttcgcgata caggacccc aaacctgcc attagggag agcgctgtg gaggctttg agggccgct ttggtcgtc acagcgcaa cgagagacc gccaaacag aaccaaaag ccacaatgg tacaaacaa tgttacgac gaatccgtg aaaatgctc tcggtggaa agaggcgaa cgcatgaaa gaggagcgc gtccacggc tttctcctc attctcaac gaactggtg cgttgctca aacgccgaa tgggaacgc aaatgcaaa agtttgctc gaacgcact gacaccaaa caaccggtc gagaccagc ttcaatttt actaaatct cgcttcgtt ttttcggtc tctcgacgc atcccccaa taccaagct gtgtctttg gcaccctct caaatcgca atcatagaa tcccaaagt tgccgaggc ctcgtttcc gagaaattc gactacatt tgtttagac gtcctggct cagagactc tcccgttcg tctcacatc caacaaagt tttctcacg attttaccc cgattggct gcttttgac cgcgaaaac tacgtcaag cagcactta agccccgtc atgaaccac atcttcaac attttgaag agtcgggaa aaggagaga gctaaggct tttgtgact attggctta attgcgatc gctgtcgag agcgacatt gagccgtac attgaacgt attatggaa ataattcgc atgtatttg ccccgagcc gatattttg gggcgcaag cggagcccc atcgactcc tcaattttc aactgtgtc actttccta gcccacgcc ctcaaaaac cactcaaaa atggacatc ccgaacctc ctggaacca atgctagca acgggatta accgcatcc ctcacgata tgtttccgg gaattatcc cgaaaagtc ccggaacat aaagaacga atcagtctc gggttatta aaaatgctg agttatatt ctcctgaat aaaccacta gttcatccc gggatgccc cgccacttg accggaacc gtaatgagt cttgcaata ccagaagtc aacgatgtg caaatcata gttttggcc ctcaaaaca ctcggaacg tttgacttt gagggccag agactcctc ccttttgtc caccgctgc gccaatcat ttcctcatc catgacaat aacgaaatc cgattggag gctgtcaga accacgtgt cggctcctc cgacatgct attcactcc acagcgaaa aactcgtcg gatacggtc accaaaacc gtagctgct gttttacat cgtttgtta agtgttggt ttgaccgat accgaccca aatgtccgg tacggagtc ttgatttcg ctcgatcgt acgtttgat aatcattta gcacaagct gagtcgctc agtgcgctg tttctggcg ttgcaagac gaaatgttt gaaatacgc gaagttgca ctgtttaca atcgggcgc ttgagtgcg atgaatccg gcatatgtc atgccctcg ttgaggaaa actttagtg cagttttta tccgaattg gaacattcg ggcagtggg aggaataag gaacaaggg gcgaggatg ttagatcat ttggttgtg agcgccccg aggatgata aggccctac atggagccc attttgaag gtcctggtg cccaaactg cgcgacccc gagccaaac cccggcgtc gtgctttcc gtcctgctc acaatcggc gatttggcc gaggtgagc agcggaggc accgaactg caagaatgg atgcaagaa ctgatgggt attttactc gaaatgttg ggcgacgct tccgccccc gaaaaacgg ggcgccgcc ttatgcacc ttgggccag ttggttgga gtgaccggt cacgtaatc caaccctac accgaatat ccaatactg ctcgacgtc ctcctcaat ttcctcaaa accgaacaa cagtcctac attcggcga gaaacaatc cgcgttttg gggctgttg ggggctttg gatccgtac aaacataaa ataaatcgg ggggagatt gattattat cccgaagcg cccgttttg ataccgatg acggataaa ggggaggat ttgaacgcg gatttgacg tcgagcgaa atgctggtt aatatgagt tcgagcact ttggaggag tactatttg gcgatagct atagccact ttgatgcgg attattaag gatccgacg ctggcacaa caccataca atggtggtg caagcagtt actttcatt tttaaaagt ttggggatt aagtgtgtg ccgtatata tcgcaagtt ttgcccagt cttttgcac gtagtacgg acagccgac gttaacttt aaggaattc ctcttccaa cagttggca caactaatc tacatcgtc aagcaacac attcgtaac tacttggac gacatttgc ctactaatc aaagaattt tggaccccg aacagtacg attcaaggg actttgatc cttttagtt gaacacatc gcagtggcg ttgggggcg caatttaag gtttattta ccgaaaatg ttgccccat atcttacgc gtgttgaac cacgacacg tccaaggaa cgactctac actatcaaa ttattggaa gcgttacac aatttcggt aacaatttg gacgaatat atgcacttg attttgccc cccattgtc cggttattc gacgctcag gagtgtcca atagtcgtg tcgaaaaaa gccctcgag actatcgac caattggcc gaaattatc gacttttcg gactttatc tcacgcatt gtccacccc ttggttcga acaattgat aactgtccc gaattgaga cccacagcc atggaaact ttatgttca ctcatgcaa caattgggg cgaaaattc tcaattttc gtacctttg gtacagaaa gtgatgacc aaacataag atacaacac agtaaattc gatacttta gtttcgaaa atccaatac gagacaaca ctggccgaa gacgttgat ttcccaatg ccgagatcc aaaacaacc gggaaaaat cgagaccca gcgatgcct gccgatagt ggaatgatt caaagactc aaagtttcg gaatccaac ctacaacag gcctggacc cctgtccga agagtctcg aaagacgat tggttggaa tggctaaga cgcctcagt atcgaacta ttaaaacag tcaccaatt ccagcatta cgttcgtgc cttacttta gcacagacg tattcgcaa ctgccacgt gatttattc aacgcagcg ttcgtaagt tgttggtcc gaattgagc gaaaacatg cagaatgaa ctgatcagt tgtctcgag caagctttg actgtgccg gacgttccg gaaataact caaactatt ttgaatttg gccgaattt atggaacat tgcgacaag gggcctttg cccctcgat agccacatt ctggggcac catgcgatg cattgccgg gcgtacgcc aaagccttg cattataag gaagaggag tttcaaagg ggggcgagc agtcaggtt gtcgaagca ctcatttcg atcaataat aagttgcaa caaaaggag gcggctgaa gggttgtta caatacgtc atgcagagg gaaatgcaa gtgcaggtc aggtggtac gagaagttg cacaattgg gagaaggcc ctgaggttg tatactgag aaactggaa ggggacgag gcggaccaa gaggcttgt ttgggcaaa atgagatgt ctcgaagcg ttgggcgaa tggggcgaa ctgcaccaa gtcgtcgag aaaagtttc agtttacta aatgacgac agcaaacaa aaggcgagt cggctagcg gctgcggcc tccttcggt ttgcacgat tacaggtca atggaaact tacgttaat gtaattcca agggatacg caagagggt gcgttctac cgtgcaatt ttggcaata cataaagag gattacgaa gtggcccaa agattcatc gattcagct cgcgattta ctcgataat gaattaact gcgatggct ggtgagtcc taccaaagg gcctacggg gccatggtc atggtacaa atgttatcc gagttggaa gaagtaatg cagtatcgg ttagtacca gaacggagg cccacttta aaagccatg tggtggcaa aggctccaa tcgggccaa aaactggtc gaagactgg caaagaatc atccaagtc cattccttg gtcttgacc ccacaagaa gacaaacga acgtgggtg aaatacgcg tcattgtgt cgcaagagc ggttcatta atgctatcg caaaaaacc ctagtcatg ttactcggt tacgaccca agcgaacgt cccgacgcc cccttgccc aaaaaccag cctcacatc acactagcc tacgccaaa catttatgg gtggcacag gaaaaacaa aaagcgttc caaaaacta tcacaattc gtcgctgac tattcgcaa aacgagcca aacgacgac gtgacagtt gaagagaaa aaaagactc ctggctagg tgttacctc aaactgggg gcctggcat gaggcactt gaaggcatc aacgaaacc tccatacct ttcattttg aagtgttac aaacaagcg actgaacac gaccctcaa tggtacaag gcatggcac gcttgggcc tatatgaat ttcgaaacg gtcttgtat tacacgaga caggaggac aagacgcac tatacagtc ttagccgtt cagggattt ttcaaatcg attaattta tctaaagga agttcattg caagacact ttacgttta ttaacactg tggttcgat tacggcgat tggcccgaa gtttacgat gccatagtc gaaggcatt cgtttagtc gagaaaaac acgtggctt caagtaatt ccgcaatta atcgcgcga attgacaca acggcgtta gtatcgaaa ttaatcaat catctattg gttgatatt ggaaaaacg catccacaa gccctagtc tatccgtta accgtcgca acaaaaagt aattcaatc agacgtaga aatgcagct aatagtatt ttaaaatcg atgagtgaa cattcccct acgttggta agtcaggcc atgatggcc tcagaagaa ttgatccga gtggcaatt ttgtggcac gaaatctgg cacgaggga ctagaagaa gcctcaaga ttgtatttc ggcgaacgt aacgttaaa ggaatgttg cgaattctc gaaccgttg catgcgatg atggaaagg gggccccag acgttgaaa gagacgagt tttaatcag acgtacggc agggatctg aacgaggcg caagactgg tgccaaaga tacaagctt tcgagcaac attagagac ctgaaccag gcgtgggac ctctactac cacgttttc cgccgcatt tcgcgtcaa ttaccgcaa ctcacctcc ctcgaatta caatacgta tcgccaaat ttgctagtt tgtcaagac ttagagttg gcagtgcct ggtagctac tgtccagga cagccaatc gtacgcatt gccaatttt aaccgatct ctcgaagta attacctca aaacagcgg ccccgaaaa ctagtaatt cgtggaagc aacggcaaa gactacatg tttttactt aaaggtcac gaggatctg agacaagac gagcgtgtt atgcaactg ttcggttta gttaatact ttactaatg aaagacccg gatacgttc aggcggaat ttgacgata cagcggtat gcggtgatt cccctgagt acgaattcg ggtctgatt gggtggtta ccgcactgt gatacgctg cacacttta atacgggat tatagagac aagaagaag attttgttg aatattgag cataggatt atgttacgg atggcgccg gattatgat catttaacc gtgatgcag aaaatggaa gtttttgag cacgcactt gagcacaca catggtgac gatttagcg cggttgttg tggctgaaa agcccctcg tctgaggtt tggtttgac cggagaacg aactacacg cggtcttta gctgtgatg tcaatggtt gggtatatt ttggggttg ggggatcgc cacccctcc aatttaatg ttggacaga ctgagcggc aaaatcctg catatagat ttcggcgat tgcttcgaa gtggcaatg actagggaa aaattcccg gaaaagatc cccttccgt ttgactagg atgctaata aacgccatg gaagtgaca gggattgaa ggcacgtac agacggact tgtgagtcg gtcatgtcc atgttacac cgaaataaa gacagtctg atggcggtg cttgaagcc tttgtttat gaccccctg cttaattgg cgattgatg gatattgag gtgcggaac ttgccgatg aacagcgat tcaatgtcg acgagttgt tcccaaaaa cagggctca tccgaatcg ctcaatgtt ccgaaacgc gagatgata atggaagat aacgaaccg caacacgaa gttaatgcc agggcggtt attatcatc aatcgagtt agagataaa ctgaccggg aacgatttc gctacggag gaacctttg actattcct aagcaagtc gatctgtta atacaacaa gcgacaagt aacgagaac ttgtcacag tgttacata ggctggtgc cccttttgg taactcgac tgctgcatc taataccgt gatagtttt ttgttgtgt atcttatta ttttttata taatctagt tgaaataaa gtaattatt acaagtgtg tgtttagga gcgcaataa acaactaag attaat
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