| General information | |||
| TC004709 | 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_007416.3: 909579..924728 | |
| 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 | TC004709 | ||
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_03819 | 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)
eclosion not fulfilled (with pupal and adult features) - Number of animals showing the phenotype: 3 (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: 30.0% (includes death as larva, prepupa, pupa)
Lethalities 22 days after larval injection: 80.0% (includes death as larva, prepupa, pupa, adult)
Days post injection
eclosion not fulfilled (with pupal and adult features) - Number of animals showing the phenotype: 3 (16 dpi)
Days post injection
adult lethality - Number of animals showing the phenotype: 2 (19 dpi)
Days post injection
adult lethality - Number of animals showing the phenotype: 2 (41 dpi)
Days post injection
- >TC004709 RA
MCQCPTHCK QKVDLVCGT DGKTYANRC QLRVASCKA RLNTRVKHD GECEQNDPC RDKQCGFGA RCVVSPDGR NASCVCPDK CPSYGDHTT SRPVCGSDG IDYRNQCEL QKAACTSNT NITIKFLGK CDPCAGVEC AEPEVCQLD EHRNPVCRC GDSCPLEFT PVCGSDGKT YSNECTLRQ EACRARKNL HIIYRGKCS SGINPCMSV RCTLGEECA INKFGIARC QCPPSCEPI MRPVCSKDG RTFPSECEL KRTACLTRT TIEISYSGV CGEKGPCSE YQCQNGATC VERFGVAHC ECPVCPAEF EPVCGSDGI SYGNECKLR LEACKHPRN ITVLYDGPC NGCETKKCD FYSVCERDS ATEGRCVCP KSCTDPELN DGTVCGTDG VTYANECEL RMTSCKMKQ FILVAYKGN CDLCQGVEC KYGGRCEAG KCVCPTNCE GSGDEPVCA SNMMTFPNE CELQKAMCL QPTNSPPLS VVFYGDCRE KFPVAGSLS TTLSPFTAQ TNNIGPGTD LDTVGPENT IPDLSTVEK EACRDIHCD FEATCELGP DNFPRCTCQ FDCASAALV SASKPVCAS DLQIYPSLC AMKMEACQR QEELRLRPL DLCQGMEVK PCNGEKPLT DPLTGKELD CGSGPKRQD CPSGSYCHQ TTRFSRCCP KDQGLLEQK SCEDSWHGC CPDGKTAAQ GPNHAGCPS LCGCNKLGS YSDTCDPET QQCRCRPGV GGAKCDRCE PGYWGLPKI SSGYHGCIP CGCSTFGSV RDDCEQMTG RCVCNPGVQ GQKCTVCTS HDKVLGPNG CVPADLTTS PPTTCKELT CYFGATCVE RGGFAICEC HTECPQEND AQVVCGSDG QTYASACEL RQVACRTQK DIVVQAFGT CKDDMFPST DWPIRRYTP LQFTQPDDS NSPLSKSTR HLLVPDPRY YYGSSGSIM RPVQGVSYP AETDNKAAF EINVSATGP NAIYNTRGG NYAPAYRPT PATVRVITA LLGDLCSDN SDCMIMYSH CVSGACTCL PNHSESSDR QECIADPNP TDEYRACSS SPCHHGSTC VDLPAATFT CVCETNFTG SLCETEVIH KQYNTPAFH GRSYVKLKP LKAYHKLSI EVEFKTHSH DGLLLYNQQ KPDGLGDFV SLAIVNGFV EFKYNLGNG HVLIRSVDK IQLGVFHRV VIKRYHRDG ILKLDEGED VAGQAKGTL KALDLLEDT YVGFLPSNY TRVYENIGT AKGFRGCIR KLKIGRRPI ELHVNRDDW VLGAEAVSE CGDSPCSAA PCANDAKCA LITPETYRC ECGPHYRGE FCEKPIEPC RSNPCKFGS TCTPLSPDA YICECPPGR TGTRCEMED KYMGIVNPE FSGSSYIRL PRLEGVRKT FSIEVYFMP KAANGLILY NGQLKNGRG DFISLNLAR GHLQFRFNL GSGIANLTT KETVNIGKW HWARIFRDG REGILQLDN SSIVRGYSG TPLTELNLE LPFYIGSLS EWDEVHRLS GASKGYKGV IQRILLNGK PLPISAKLP DCSFDSNRN GCAFNVGSY DGLPCPISK NPCLNNGLC VPELDQFIC KCPSNTKGK YCEISQEET PAIKFNGAT FLQFRNRGY RSHNLTISD EYPDNEEDD IDNIEEDVV DNNTYDAYD DEEYYFDFY DTARRKPER GNRYEIKLR TFASDALLL WRSKSRSIR EDYFSIAVV NGYPEISYN LGKQDVFWA IRSKTKIDD GKWHTIQVR RRKRVGFIS IDGEPPIKG LSKYGAISL RTNSKLWIG GTANLPQGL PSAYYKGFE GCIQHILVN AKPLDMVSN NELNKVNFC HDNEI
- >TC004709 RA
atgtgccaa tgtccaacc cattgcaag cagaaggtc gacctcgtc tgcggaact gatggaaag acttacgct aatcgctgt caacttcgt gtggcatct tgcaaagct cgacttaac accagggtt aaacatgat ggggaatgc gaacaaaac gatccatgt agagacaag caatgcggt tttggggct cgatgtgtg gtttcccct gatggccgc aatgccagc tgtgtctgt ccggataaa tgccccagc tatggagat cataccacc tcgcgtcca gtctgtgga agcgacggg atcgactat cgaaaccaa tgcgagctg caaaaagcc gcctgcact tccaacaca aacatcacc atcaagttt ctcggaaag tgcgatcct tgtgccgga gtggaatgt gcagaaccc gaagtatgc cagttggac gaacatcgc aatcctgtt tgtagatgt ggggacagt tgtccttta gaatttact ccggtttgc ggatcggac ggaaaaacc tactccaat gagtgcacg ctgcggcag gaagcctgc agggcccgg aaaaacctc catatcata tatcgcgga aaatgcagc tcaggcata aacccgtgc atgagcgtg cggtgcacc ttaggggag gaatgtgcg ataaataaa ttcggcatc gcccgctgt caatgcccg ccgtcttgc gagcctatc atgaggccc gtttgttcg aaagacggc cgaactttc ccatctgaa tgcgaattg aaaagaacg gcttgtctg actcggacc actattgag atatcttac agtggcgtt tgcggggag aaaggcccg tgtagcgag taccagtgt caaaatgga gcgacctgc gtcgaacga ttcggtgtt gcccactgc gagtgtccc gtctgtccg gccgaattc gaacccgtc tgtggatcc gacggtatt tcgtatggc aacgaatgt aagctaaga ctggaagct tgcaagcat ccgcgcaat attacagtc ctttacgac ggtccttgt aacggttgc gagactaaa aaatgcgat ttttactcc gtttgcgag cgcgatagt gcaactgag ggacgttgt gtttgcccc aagtcatgt acggaccct gagctcaac gacggaacg gtttgtgga acagatgga gtgacttac gcaaacgaa tgcgaactt cgcatgacc tcttgcaaa atgaagcag ttcattttg gtcgcgtat aagggaaac tgcgatcta tgtcaggga gtggaatgt aaatacggg ggtagatgc gaagcgggc aaatgcgta tgtccgacg aattgcgaa ggttccggc gatgagccg gtgtgtgca tcaaacatg atgacgttc cccaacgag tgcgaattg cagaaagcg atgtgttta cagccgaca aattctccc cccttgagc gtagtgttc tatggcgac tgtcgcgag aaattccca gtagcagga tctctcagt acaactctt tcccccttc acggctcaa actaacaac atcggcccg ggcacggat ctagacact gtcggtccg gagaataca attccagac ttatcgacg gtcgagaaa gaagcctgc cgtgatatt cattgcgat ttcgaagcc acgtgcgag ttaggtcca gacaacttc ccacgttgc acctgtcag ttcgattgt gccagtgcc gcactcgtc tctgcctcg aaaccagta tgcgcctcc gatttgcaa atctatcct tcgctttgc gctatgaag atggaagcg tgtcaaagg caggaggag ttgcggttg aggccactc gatctctgt caaggaatg gaggtaaaa ccatgcaat ggcgaaaaa ccattaaca gaccccctc acaggaaag gaattagat tgcggaagc ggcccgaaa agacaagat tgcccttcc ggcagctac tgccatcaa accacgcgg ttttcccgt tgttgtcca aaagaccag ggcctttta gagcaaaag agctgcgag gatagctgg cacgggtgt tgtcctgac ggaaaaacc gccgcccaa ggccctaat cacgccgga tgtccttcc ctctgcggc tgcaataaa ctcggctca tattccgac acgtgcgat cccgaaacg caacagtgt cggtgtaga ccgggcgta ggaggtgcc aaatgtgat cgatgcgaa cctggatat tggggttta ccgaaaatc tcctccggc tatcacggt tgcatcccg tgcggttgt tcgacgttc ggctccgtg cgagacgat tgcgagcag atgacgggg agatgtgtg tgcaacccg ggggtgcaa ggccaaaaa tgcacagta tgcaccagc catgacaag gtcttagga ccgaacggg tgcgtacca gcggatctg actacgtcg cccccgacg acgtgcaaa gaattgact tgttatttt ggagcaaca tgcgtcgaa cggggtgga tttgctata tgtgaatgt cataccgaa tgccctcaa gaaaatgac gctcaagtc gtgtgtgga agtgacgga cagacgtac gcgtcggcg tgcgagttg cgacaagtc gcatgtcgg acgcaaaaa gatatagtc gtccaagcg ttcgggacg tgtaaagat gatatgttt ccaagtacg gattggcca attcgtcgt tatactcct ctccagttt acacaaccc gacgattct aattcgcct ttatcaaaa tcgacacga catttacta gttcccgat cctcgttac tactacggg agcagcgga tctattatg cgacccgtt caaggagtc tcttatccc gctgaaaca gataataaa gccgcattc gaaataaac gtatctgct acaggacct aacgccatt tacaatacc cgcggtgga aactacgcc ccggcttac cgtccgacc cccgcaacc gttcgcgta ataacagcc cttctcggg gatctctgt tccgataac tccgattgc atgataatg tatagccac tgcgtatca ggagcgtgc acttgtctt ccgaatcac tctgaatcg tccgaccga caagagtgc atagctgat ccgaatccg accgatgaa tatcgtgca tgtagctca tccccctgt catcatggt agcacttgt gttgatctt cccgccgcc acgttcact tgcgtttgc gagacaaac tttacgggg tctctttgc gaaaccgaa gtcattcac aaacaatac aacactcca gctttccat gggcgttct tacgtcaaa ctcaaacct ctcaaagcc taccacaaa cttagcatc gaagtcgag ttcaaaacg cattcccac gacggattg ctgctctat aaccagcag aaacctgac gggttgggc gatttcgtt tctcttgcc atcgtcaac ggatttgtc gaattcaaa tacaatcta ggcaacgga cacgttctt atccgttca gtcgacaaa atacagctc ggcgtcttc caccgagtc gtcattaag agatatcac agagacgga atactcaaa cttgatgaa ggggaagac gtggccgga caggccaaa ggcacactc aaagctctc gatttactt gaagataca tacgtcgga tttctaccc tctaactac acaagagtt tacgaaaat atcggcacc gcgaagggt tttcggggc tgcatcaga aaattaaaa attggacga agaccgatc gagctgcac gtcaacaga gacgattgg gtcttgggc gctgaggcg gtatccgag tgcggtgac agcccctgc tcggcggcg ccctgtgcc aacgacgcc aagtgcgcc ctaattaca cctgaaacg taccgatgc gaatgcggc cctcattat cggggcgag ttctgcgag aagccgatc gagccttgc agatccaac ccttgcaaa ttcggatcg acgtgcacg ccactcagc cccgacgct tacatctgc gaatgtcca cctggacgg accggaact cggtgcgaa atggaagat aaatatatg ggtatagta aatccggaa tttagtggg tcaagttat atccgactt cctcgatta gagggtgtt aggaagacg ttttcgatt gaggtgtat tttatgcca aaggcagca aacggtctg atattgtat aacgggcaa ctgaagaat ggccgcggt gattttatt tcgttgaac ttagcacgt ggtcattta caatttaga ttcaatttg ggaagcgga attgcgaat ttaacaact aaagagact gtaaatatt gggaagtgg cattgggcc cgtattttc cgagacgga cgagaagga attttacaa ttggataat tctagcata gttcgtggt tattccggc acgccacta acagaactt aatctggaa ttgccgttc tacatcgga tctttatcg gagtgggat gaagtgcat cgtttatct ggcgcatct aagggatat aagggcgta atacaaagg attctccta aatggaaaa ccccttccc atttcggct aaattgccg gattgctct tttgattct aatagaaac ggatgcgca ttcaatgtc ggctcatat gacggatta ccatgtccc atttccaaa aatccttgc ctaaataac ggcctatgt gtaccagaa ctcgatcag tttatatgc aaatgccct tctaacaca aagggaaaa tattgcgag atatctcaa gaagaaacc cccgctatt aaattcaac ggtgccact tttctccaa tttaggaac agaggatac agaagtcac aatttaaca ataagtgat gaatatccc gacaacgaa gaggacgat attgacaat attgaagaa gacgtcgtc gataataac acttacgac gcttacgac gacgaagaa tactacttc gatttttac gataccgct agacgaaaa ccggaacgt ggcaacaga tacgaaatc aaattgcgc acctttgcc agcgacgct cttctcttg tggaggagt aaaagtaga agtataaga gaagactat ttttctata gctgttgtt aatggttac cctgaaatt agttataat ttaggcaag caagatgtc ttctgggca atcagatcg aagacgaaa atcgacgac ggcaagtgg catacaatc caagtaagg agacggaaa cgagtaggt tttatatca atcgacggg gagccgcct attaagggc ctctcgaaa tacggtgcc atttcactg agaacgaat tctaaatta tggataggt ggtacggcg aatctgccc caaggccta ccttcggcg tactacaaa ggcttcgaa ggatgcatc caacatatt ttagttaac gcaaaaccc ttagatatg gtttcaaat aatgaatta aataaagtt aacttctgc cacgataac gaaatttaa
- >TC004709 RA
ttggggggc ccagtgcat caccggccc cgacggcag ggccatgtg ccaatgtcc aacccattg caagcagaa ggtcgacct cgtctgcgg aactgatgg aaagactta cgctaatcg ctgtcaact tcgtgtggc atcttgcaa agctcgact taacaccag ggttaaaca tgatgggga atgcgaaca aaacgatcc atgtagaga caagcaatg cggttttgg ggctcgatg tgtggtttc ccctgatgg ccgcaatgc cagctgtgt ctgtccgga taaatgccc cagctatgg agatcatac cacctcgcg tccagtctg tggaagcga cgggatcga ctatcgaaa ccaatgcga gctgcaaaa agccgcctg cacttccaa cacaaacat caccatcaa gtttctcgg aaagtgcga tccttgtgc cggagtgga atgtgcaga acccgaagt atgccagtt ggacgaaca tcgcaatcc tgtttgtag atgtgggga cagttgtcc tttagaatt tactccggt ttgcggatc ggacggaaa aacctactc caatgagtg cacgctgcg gcaggaagc ctgcagggc ccggaaaaa cctccatat catatatcg cggaaaatg cagctcagg cataaaccc gtgcatgag cgtgcggtg caccttagg ggaggaatg tgcgataaa taaattcgg catcgcccg ctgtcaatg cccgccgtc ttgcgagcc tatcatgag gcccgtttg ttcgaaaga cggccgaac tttcccatc tgaatgcga attgaaaag aacggcttg tctgactcg gaccactat tgagatatc ttacagtgg cgtttgcgg ggagaaagg cccgtgtag cgagtacca gtgtcaaaa tggagcgac ctgcgtcga acgattcgg tgttgccca ctgcgagtg tcccgtctg tccggccga attcgaacc cgtctgtgg atccgacgg tatttcgta tggcaacga atgtaagct aagactgga agcttgcaa gcatccgcg caatattac agtccttta cgacggtcc ttgtaacgg ttgcgagac taaaaaatg cgattttta ctccgtttg cgagcgcga tagtgcaac tgagggacg ttgtgtttg ccccaagtc atgtacgga ccctgagct caacgacgg aacggtttg tggaacaga tggagtgac ttacgcaaa cgaatgcga acttcgcat gacctcttg caaaatgaa gcagttcat tttggtcgc gtataaggg aaactgcga tctatgtca gggagtgga atgtaaata cgggggtag atgcgaagc gggcaaatg cgtatgtcc gacgaattg cgaaggttc cggcgatga gccggtgtg tgcatcaaa catgatgac gttccccaa cgagtgcga attgcagaa agcgatgtg tttacagcc gacaaattc tcccccctt gagcgtagt gttctatgg cgactgtcg cgagaaatt cccagtagc aggatctct cagtacaac tctttcccc cttcacggc tcaaactaa caacatcgg cccgggcac ggatctaga cactgtcgg tccggagaa tacaattcc agacttatc gacggtcga gaaagaagc ctgccgtga tattcattg cgatttcga agccacgtg cgagttagg tccagacaa cttcccacg ttgcacctg tcagttcga ttgtgccag tgccgcact cgtctctgc ctcgaaacc agtatgcgc ctccgattt gcaaatcta tccttcgct ttgcgctat gaagatgga agcgtgtca aaggcagga ggagttgcg gttgaggcc actcgatct ctgtcaagg aatggaggt aaaaccatg caatggcga aaaaccatt aacagaccc cctcacagg aaaggaatt agattgcgg aagcggccc gaaaagaca agattgccc ttccggcag ctactgcca tcaaaccac gcggttttc ccgttgttg tccaaaaga ccagggcct tttagagca aaagagctg cgaggatag ctggcacgg gtgttgtcc tgacggaaa aaccgccgc ccaaggccc taatcacgc cggatgtcc ttccctctg cggctgcaa taaactcgg ctcatattc cgacacgtg cgatcccga aacgcaaca gtgtcggtg tagaccggg cgtaggagg tgccaaatg tgatcgatg cgaacctgg atattgggg tttaccgaa aatctcctc cggctatca cggttgcat cccgtgcgg ttgttcgac gttcggctc cgtgcgaga cgattgcga gcagatgac ggggagatg tgtgtgcaa cccgggggt gcaaggcca aaaatgcac agtatgcac cagccatga caaggtctt aggaccgaa cgggtgcgt accagcgga tctgactac gtcgccccc gacgacgtg caaagaatt gacttgtta ttttggagc aacatgcgt cgaacgggg tggatttgc tatatgtga atgtcatac cgaatgccc tcaagaaaa tgacgctca agtcgtgtg tggaagtga cggacagac gtacgcgtc ggcgtgcga gttgcgaca agtcgcatg tcggacgca aaaagatat agtcgtcca agcgttcgg gacgtgtaa agatgatat gtttccaag tacggattg gccaattcg tcgttatac tcctctcca gtttacaca acccgacga ttctaattc gcctttatc aaaatcgac acgacattt actagttcc cgatcctcg ttactacta cgggagcag cggatctat tatgcgacc cgttcaagg agtctctta tcccgctga aacagataa taaagccgc attcgaaat aaacgtatc tgctacagg acctaacgc catttacaa tacccgcgg tggaaacta cgccccggc ttaccgtcc gacccccgc aaccgttcg cgtaataac agcccttct cggggatct ctgttccga taactccga ttgcatgat aatgtatag ccactgcgt atcaggagc gtgcacttg tcttccgaa tcactctga atcgtccga ccgacaaga gtgcatagc tgatccgaa tccgaccga tgaatatcg tgcatgtag ctcatcccc ctgtcatca tggtagcac ttgtgttga tcttcccgc cgccacgtt cacttgcgt ttgcgagac aaactttac ggggtctct ttgcgaaac cgaagtcat tcacaaaca atacaacac tccagcttt ccatgggcg ttcttacgt caaactcaa acctctcaa agcctacca caaacttag catcgaagt cgagttcaa aacgcattc ccacgacgg attgctgct ctataacca gcagaaacc tgacgggtt gggcgattt cgtttctct tgccatcgt caacggatt tgtcgaatt caaatacaa tctaggcaa cggacacgt tcttatccg ttcagtcga caaaataca gctcggcgt cttccaccg agtcgtcat taagagata tcacagaga cggaatact caaacttga tgaagggga agacgtggc cggacaggc caaaggcac actcaaagc tctcgattt acttgaaga tacatacgt cggatttct accctctaa ctacacaag agtttacga aaatatcgg caccgcgaa gggttttcg gggctgcat cagaaaatt aaaaattgg acgaagacc gatcgagct gcacgtcaa cagagacga ttgggtctt gggcgctga ggcggtatc cgagtgcgg tgacagccc ctgctcggc ggcgccctg tgccaacga cgccaagtg cgccctaat tacacctga aacgtaccg atgcgaatg cggccctca ttatcgggg cgagttctg cgagaagcc gatcgagcc ttgcagatc caacccttg caaattcgg atcgacgtg cacgccact cagccccga cgcttacat ctgcgaatg tccacctgg acggaccgg aactcggtg cgaaatgga agataaata tatgggtat agtaaatcc ggaatttag tgggtcaag ttatatccg acttcctcg attagaggg tgttaggaa gacgttttc gattgaggt gtattttat gccaaaggc agcaaacgg tctgatatt gtataacgg gcaactgaa gaatggccg cggtgattt tatttcgtt gaacttagc acgtggtca tttacaatt tagattcaa tttgggaag cggaattgc gaatttaac aactaaaga gactgtaaa tattgggaa gtggcattg ggcccgtat tttccgaga cggacgaga aggaatttt acaattgga taattctag catagttcg tggttattc cggcacgcc actaacaga acttaatct ggaattgcc gttctacat cggatcttt atcggagtg ggatgaagt gcatcgttt atctggcgc atctaaggg atataaggg cgtaataca aaggattct cctaaatgg aaaacccct tcccatttc ggctaaatt gccggattg ctcttttga ttctaatag aaacggatg cgcattcaa tgtcggctc atatgacgg attaccatg tcccatttc caaaaatcc ttgcctaaa taacggcct atgtgtacc agaactcga tcagtttat atgcaaatg cccttctaa cacaaaggg aaaatattg cgagatatc tcaagaaga aacccccgc tattaaatt caacggtgc cacttttct ccaatttag gaacagagg atacagaag tcacaattt aacaataag tgatgaata tcccgacaa cgaagagga cgatattga caatattga agaagacgt cgtcgataa taacactta cgacgctta cgacgacga agaatacta cttcgattt ttacgatac cgctagacg aaaaccgga acgtggcaa cagatacga aatcaaatt gcgcacctt tgccagcga cgctcttct cttgtggag gagtaaaag tagaagtat aagagaaga ctatttttc tatagctgt tgttaatgg ttaccctga aattagtta taatttagg caagcaaga tgtcttctg ggcaatcag atcgaagac gaaaatcga cgacggcaa gtggcatac aatccaagt aaggagacg gaaacgagt aggttttat atcaatcga cggggagcc gcctattaa gggcctctc gaaatacgg tgccatttc actgagaac gaattctaa attatggat aggtggtac ggcgaatct gccccaagg cctaccttc ggcgtacta caaaggctt cgaaggatg catccaaca tattttagt taacgcaaa acccttaga tatggtttc aaataatga attaaataa agttaactt ctgccacga taacgaaat ttaaatgat gtaaattat gttgcacgg cgcaatatt aaataattt aaaactgtg agactgata tattgtact cctcaattt tccatttac agaatttag cttagagat gatgttgta tataaatgt actgttaat aaattacag cagttttat gtagcaact ttgaaacat tggctgcca gtacaataa ttacctact aacatatca aacgggaag tttttactt aatttttca aatgtcttc agttgtaag cattagaga ctggaaaca ttggagcca attaaaagt gtacaaata ttttttgtt cctagtagt tgtaatttg gattaaaac atatctttc gaagttggc agtcaatgt attaaatag acacggtaa ataggagtg tttttacaa agctgtttg tatctagtc ttgtattta tttccgaat taggtcttg gattttgta aaggacgac gtaaaacct gccatttct aagttctcg tggactgac actttatcg atagttgag gctttaatt tgatactta taatgaatg cttaaataa gaaatgtgg actaaaatt gtttgtatg gccaatgtt gaaatttat tatcttgaa aatgtttag atgttaccc gattccagt aacaactgt gtgaacttc atatctgta actactgat taagtcgaa ttacttgtt ttctggcaa taattttta tggtacata agaggagtt aatatgctt gtatgttat gtaaatagc caatacgaa atgatagtt attttaggt aatgttaat ttatagtaa aaattttat tacgtaatg aatagattg atgagctaa attaattat aatgataag ttttgttta gtatactga ttgtttcta ttgtagacc tctgtacta taaatgcta caaaatata tttttttcc tacta
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