| General information | |||
| TC014935 | 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: 7879159..7909931 | |
| 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 |
|
TC014935 | |
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_02825 | 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: 50.0% (includes death as pupa, adult)
eclosion not fulfilled (with pupal and adult features) - Number of animals showing the phenotype: 5 (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: -1.0% (includes death as larva, prepupa, pupa)
Lethalities 22 days after larval injection: -1.0% (includes death as larva, prepupa, pupa, adult)
- >TC014935 RA
MARDDKLFF HKTVQHLAR SLACIKNTP WDKVKTLYD LCPIETTNG TVSLNSRQQ DAVIALGIY FLESGLEHK DTILPYLLK LAKALGKAT WLDEIKQNP SDRIPVAEK FSFCLHTLL CDIAVKCED SREEIIETQ VDCLVKLTN SILKTRENT NSAVKLFLC KTTVPVLIG LSRAMGRFC NTEPPLICR LFPKPEPPL SPVTSNPDY KRSFSNFRS IIPRSLSGN LAATVDILA ITQGYDTTD VAYSSASLK RGSLINQNF VNYDPATYF FSKFGSSFN QFPHLRVND PNDKKGQII FPLQHLQTI LSLAKKLLT KDMLSFLDE QSLEVYTTG KIVIFPYKT FSETINLVM VTLMRELLQ PQKSLPVAF TKDVQEFVK GLYLNGQTE LQSRNHDAS EKEDRESNF ALVNKFKVN VMANAACVD LLVWAIGDE TGADSLCGR LTEKINSNH GYKLVLAHM PLVMVCLEG LGTLAQKFP NIASTSIYC LRDFLITPS PILSKLHRQ ANEKGNKEN LKIIAHVNG LKAETTKPH NPTQSAFEK LRDAAIENL CYALEAAHT TDADCVRAL VASVSNRLF RAEKSDSES TLISTNIVI MLGHVAVSM KGTPKTTDT ILQFFQQRF CRVPSALDT LIVDQLGCM IIAQCEPHV YEVVMKMFT MITVESSNA AYGNPTNEK AQYRHVSRP VINALANIA ANIQSESQM NELLGRLLE LFVQLGLEG KRASEKSPG ALKASSSAG NLGVLIPVI AVLLRRLPP IKNPKPRIH KLFRDFWLY CVIMGFTAS DSGLWPKEW YEGVKEIAV KSPALVSPT SSRSEMREL QYTSAVRND SVSITELQE LKNQILELL KQPADVTAY VNRLTFAQC TFLLSVYWV EILRIQNSP EPSLVPIIT EYLSDSALQ KDKSGMWVC VSAVGERVF EKFLEVMKN KPKYEAREA ELEGHAQFL LVHFNDPHK QIRHVSDKF LVSLFDRFP HLLWSRKVL WTMLDIMQV LSNSLHLDP NQETPTLRI PRTPYSIQL MDTLEARET KVKHFAANS ERIIKEALK WAPHWTRSH IQEYIINQG QGAGLWNHT GLSLALETI LQFGPLNMS SAPMSVSTL EKRPKCVKS DSSKLMVST SLRCKYIGE VIGLSAVYG TEGKEKLIE FIMKRVWTA CQERSDSEH RDALWQATA LLISTTEFH RNLLHCIAW SQVELFTVE AMRTAVECW QWLITSRPE LEIRFLQEM VSAWNCTVQ KRMGLFSTS EPMTSPLAA YEGARLEPN PPFVKPHGI WVQFICELI DNVKYSSYE KVEMLASLI HRSLAMCVG ADPPCQTRS VSAVGVRFK LLTCGLSLL QGDILPKSL AKNVLRERI YCSCLDYFC KPVMCPSQT PTELREDIT TLVRFWQTL HSDKKYLKA SDVGDLDIG QNAPMMVEN NELTKPNDF NRPTSGWIN TVPLSSSTA TLSKRSAKS KRVPMADNF VKCYLKKRN LIMDLLTVE IEFLIVWLN PTSRQEQQI PGEENIASW RAKTITEKQ WQDYTRLAW DISPVLATY LPSRFKTNE AIMSEIRHQ VQQNPVSVS HVPEALQYL ATTNAILSD STKLVHMLT WARVSPITA LAYFSRQFP PHPITAQYA VKVLSSYPA DAVLFYIPQ LVQALRHDT MGYVTEFIK YVAKKSQIV AHQLIWNMK TNMYLDEDM QHKDVVLFD VLDALCNSI LAELSGPAK QFYEREFDF FDKITSISG EIRPYPKGP ERRRACLEA LRKIKVQPG CYLPSNPEA MVVDIDYNS GVPMQSAAK APYLARFKV CRCGINELE NIAMAVSVN ENHEQNFGP EMWQAAIFK VGDDVRQDM LALQVIGIF KNIFQTVGL DLYLFPYRV VATAPGCGV IECVPNAKS RDQLGRQTD IGLYEYFLK KYGEESSKE FQNARRNFI ISMAAYSVV GFLLQIKDR HNGNIMLDT DGHIIHIDF GFMFESSPG GNLGFEPDI KLTDEMVMI MGGKMEAAP FKWFSDLCV QAYLAVRPY QESIISLVS LMLDTGLPC FRGQTIKLL RGRFHPGAT DKEAATYML QIIRNSFLN FRTRTYDMI QYYQNQIPY
- >TC014935 RA
atggctaga gatgataag ctttttttc cacaaaact gtgcaacat ttagccaga tctctagcc tgcattaaa aacacacca tgggataag gtgaagacc ttgtacgac ctgtgcccc attgagaca accaacggc actgtctcg ttaaacagt cggcagcaa gatgccgtt atcgccctc gggatctac ttcctggaa tcgggactc gagcacaag gacacgatt ttgccctat ttactcaag ctggcaaaa gcgcttggg aaagccact tggttggac gaaattaag cagaatccg agcgatagg attcccgtg gctgaaaag ttcagtttt tgtctccac acccttctg tgtgacatc gcagtgaaa tgtgaagat tcccgggaa gaaatcatc gaaacacaa gtcgattgt ttggtgaaa ctcaccaac agtatcctc aaaactcgc gaaaataca aacagtgca gtcaaactc ttcctgtgt aaaaccaca gtccctgtt ttaattggc ctgtctcgg gcaatgggg cgcttctgc aacactgag cccccttta atctgccgc ctgttcccc aaaccggag cccccttta tccccagtc acctccaac cccgactat aagcgcagt tttagcaat ttccgctca atcattccg agaagtctg tccggtaat ttggctgca actgttgat attttggca ataactcaa gggtacgac actacagat gttgcatat agttctgcc agtcttaaa aggggctca cttatcaat caaaatttc gtcaattat gatcctgca acttatttt ttctcgaaa ttcgggtcc agttttaat cagtttccg catttacgt gtcaatgat ccgaatgat aagaaggga cagattatt tttcccttg caacatttg caaacgata ctttcgttg gcgaaaaaa ctgctcacg aaagatatg ttgagtttt ttggatgag caaagtttg gaggtttat acgacgggg aagattgtt atttttccg tataaaacc ttctcggag acgattaat ttggttatg gtgactttg atgagggag ttgcttcag ccgcagaag agtctcccg gttgcgttc acgaaggat gtgcaagaa ttcgttaaa gggttgtac ttaaacggc cagacggaa cttcagagt cgcaaccac gacgctagt gagaaagaa gaccgtgaa agtaatttt gccctcgtt aataaattc aaagttaac gtgatggca aacgccgcc tgtgttgat cttctagtc tgggcaata ggggacgaa acaggtgcc gatagtctt tgtggccgt cttaccgaa aaaatcaac tcaaatcac ggctacaaa ctcgtctta gcccacatg cccctcgtt atggtgtgt cttgaaggc ttgggcact ttagcccaa aaattcccc aacattgcc tccacttcc atttactgt cttcgagat ttcctcatc actccttca cccatcctt tccaaactc caccgacaa gcgaacgaa aaaggaaac aaggaaaat ttaaaaatc attgcccac gttaacggc cttaaggcc gaaaccact aaaccacac aacccaact caatccgca tttgagaaa ctacgagac gctgctatt gaaaatcta tgttatgct cttgaggct gcacacact actgatgcc gactgtgtg agggcgcta gtggcgtcc gtttcaaat cggttgttt cgcgctgaa aaaagtgac agtgaatcg actctcatc tctactaat attgttata atgttgggt catgtggcc gtttcgatg aaaggcacc ccaaagaca acggatacg attttgcag tttttccaa cagagattt tgtcgagtg ccttcggca cttgataca cttattgtt gaccagttg ggttgtatg attatcgca cagtgtgaa ccgcatgtt tacgaagtt gtaatgaag atgtttacg atgattacg gtggagagt agcaatgcg gcgtatggg aatcccacc aatgagaag gcacagtat aggcacgtt tctcgccct gtgatcaac gcattggcc aacatcgcc gccaatata caaagcgaa tcccaaatg aacgaacta ttaggccgc ctgctcgaa ctcttcgtg caattgggc ctcgagggc aaacgcgca agcgaaaaa tccccaggc gccctaaaa gcgtcaagt tcagccggt aatttagga gttttaatc cctgtaatt gccgttctt ctacgccga ctaccaccc atcaaaaat cccaaacct cggattcat aaattgttc cgcgatttt tggctctat tgtgttatt atgggattt acagcgtct gattcggga ttatggccg aaagaatgg tacgaagga gtgaaagaa atcgcagtt aaaagccca gcgttagtt tcaccaact tccagtcgc tcggaaatg cgcgaattg caatacacg agcgccgtt cgaaacgat agtgtttca ataaccgaa ctacaagaa ttgaagaat caaattcta gaattgttg aaacaaccg gcagatgtc accgcctat gtgaatcgc ttgactttt gcccaatgt acgttctta ttgagcgtt tattgggtg gagattttg cgaattcag aattcacct gagccgagc ttggtgccg attattacc gaatacttg tcggattca gcgttacag aaagacaag tccgggatg tgggtttgt gtgtcggct gttggcgag cgcgttttt gaaaaattc ctcgaagtt atgaaaaat aagccgaag tatgaggcc agggaggcg gagcttgaa ggccacgcc cagtttctt ctcgtacat ttcaacgat ccgcacaaa cagattcgt cacgtttcg gataagttt ctggtcagt cttttcgat cgctttccg cacttgttg tggagtcgg aaggtgttg tggacaatg cttgacata atgcaagtg ttgtcgaat tccttgcat ttggatccg aatcaggaa acgcccact ttgcgaata ccacgaacg ccatactcc atacagttg atggacacg ttagaggca cgcgagact aaagttaag cattttgct gcgaattcc gagcgaatt atcaaggag gcgttgaag tgggcaccg cattggacg aggtcgcac atacaagag tatataata aatcaagga cagggggcg gggctttgg aaccacact ggtctgtca ttggcgctt gaaactata ctacagttc ggacctttg aatatgtca agtgcaccg atgagtgtc tcaacgttg gagaagaga ccgaagtgt gttaaaagt gattcgtcg aaattgatg gtttcgacg tctttaagg tgcaaatat attggagag gtaattggg ttatcggcc gtttatggt accgaaggt aaagaaaaa ttgattgaa tttataatg aaacgggtg tggacagcg tgtcaggaa cggtccgat tctgaacat cgagatgcc ttgtggcaa gctacggca ctcttgatt tccacgaca gagtttcac cggaatttg ttgcattgc attgcgtgg tcgcaagtc gagcttttc acagtggaa gcaatgcga acagccgtc gaatgctgg caatggctc ataacatca cgtccagaa ctcgagatc cgtttcctc caggaaatg gtctcggcc tggaactgc accgtccag aaacgcatg ggtcttttt tcgaccagc gaacccatg acaagcccc ctagcggcg tacgaaggc gcccgactt gaaccgaac cccccattc gtaaaaccg cacggaatt tgggtccag ttcatttgc gaattgatc gacaacgtc aaatacagc agttacgag aaagtcgaa atgttggca agcttgatc catcgctcg ttggcaatg tgcgtgggg gccgacccc ccatgccaa acccgctcc gtttcagcg gtcggtgtc cgcttcaag ctactaact tgcggctta tcgctactc caaggcgat attttgccg aaaagtctg gcgaaaaat gtgctcaga gagcgcatt tattgcagc tgtttggac tatttctgc aaaccggtc atgtgtccg tcgcaaaca ccgaccgaa ttgcgtgag gatatcacg acacttgtg cgattttgg caaacgtta cactctgat aagaagtat ttgaaagcc agcgatgtt ggcgatttg gacattggt cagaatgca ccaatgatg gtcgagaat aacgaactg acgaaacct aacgacttc aaccgacca acttcgggt tggatcaac accgtacca ctgtccagc agtaccgcc accttaagc aaacgctcg gcgaagtca aaacgcgtc ccgatggcc gacaacttc gtcaaatgt tatttgaaa aaacgcaac ctaattatg gacttgtta acggtcgag attgaattt ttgatcgtt tggctcaac ccgacttcg cgccaggaa cagcaaatc ccaggcgag gaaaacata gcatcatgg agggctaag accatcacc gagaaacaa tggcaagac tacacaagg cttgcttgg gatatttcg ccggttttg gccacttat ctgccctct agattcaaa acaaacgaa gctataatg agcgaaatc aggcatcaa gtgcaacaa aacccagtc agtgtgtcg cacgttccc gaagcgttg caatacctg gccaccact aacgccatc ttgagcgac agcactaaa ctagtgcac atgctgaca tgggcccgg gtctcccct atcactgcc ttggcctac ttcagtcga caattcccc ccgcatcct attacagca cagtatgct gtgaaagtg ttgagttcg tatccggcc gatgcggtt ttgttttat attccgcag ttggtgcaa gcgttacgc cacgatacg atggggtat gtgactgaa tttattaaa tatgtggcg aaaaaatcg caaattgtg gcacatcag ttgatttgg aatatgaaa actaatatg tatttggat gaggatatg cagcataag gatgttgtc ttgtttgat gttttggat gcgttgtgt aatagtatt ttggctgag ctgtcgggg ccagcgaaa cagttttat gaacgagaa tttgatttc tttgataag attacgagc atttcgggg gagatacgg ccctatccg aaggggcct gagcgcagg agggcctgt ctggaagcg ttgagaaaa attaaggtc caacctggt tgctacttg cccagcaac cccgaggca atggtggtc gacatcgac tacaacagt ggcgtaccg atgcaaagc gccgccaaa gccccatac ttagcccgt ttcaaagtg tgccgatgc ggaataaac gaactcgaa aacatcgca atggctgta tccgttaac gaaaaccat gaacaaaac ttcggtccg gaaatgtgg caagcggcc atatttaaa gtcggcgat gacgtccgg caagacatg ttggcactg caagtgatc ggaatattt aagaatatt ttccaaacg gtcggtttg gatttatat ttgtttccc tatcgggtc gtagccacc gcacctggg tgtggggtg attgaatgc gtaccaaac gccaagtca cgtgaccag ttagggcgg cagaccgac atcggactg tatgaatat tttttgaaa aaatacggg gaggagagt tcaaaggag tttcaaaac gcgcgccga aatttcatc atatccatg gccgcctac agcgttgtt ggctttctt ttacaaatc aaagaccgt cacaatggc aatattatg ttggacacg gacggtcat ataatccac attgatttt ggtttcatg ttcgagtca tcgccaggc ggtaacttg ggttttgag cccgatatt aagctcacc gatgagatg gttatgata atggggggc aaaatggaa gcagccccc tttaaatgg ttttcggac ctgtgtgtc caggcttat ttggccgtc aggccttat caggagtcc atcatttcg ctggtctcg ctcatgttg gacacgggg ctgccctgc tttcggggg cagacgatc aagttgtta agggggcgc ttccatccg ggggccacc gataaggaa gcggccact tacatgctg caaatcata cgtaacagt ttcctgaat ttcaggaca aggacctat gacatgatc caatattat cagaatcag attccttat taa
- >TC014935 RA
acctaatgt tttgacgtt tgaggttag gttaggtcg taatggcta gagatgata agctttttt tccacaaaa ctgtgcaac atttagcca gatctctag cctgcatta aaaacacac catgggata aggtgaaga ccttgtacg acctgtgcc ccattgaga caaccaacg gcactgtct cgttaaaca gtcggcagc aagatgccg ttatcgccc tcgggatct acttcctgg aatcgggac tcgagcaca aggacacga ttttgccct atttactca agctggcaa aagcgcttg ggaaagcca cttggttgg acgaaatta agcagaatc cgagcgata ggattcccg tggctgaaa agttcagtt tttgtctcc acacccttc tgtgtgaca tcgcagtga aatgtgaag attcccggg aagaaatca tcgaaacac aagtcgatt gtttggtga aactcacca acagtatcc tcaaaactc gcgaaaata caaacagtg cagtcaaac tcttcctgt gtaaaacca cagtccctg ttttaattg gcctgtctc gggcaatgg ggcgcttct gcaacactg agccccctt taatctgcc gcctgttcc ccaaaccgg agccccctt tatccccag tcacctcca accccgact ataagcgca gttttagca atttccgct caatcattc cgagaagtc tgtccggta atttggctg caactgttg atattttgg caataactc aagggtacg acactacag atgttgcat atagttctg ccagtctta aaaggggct cacttatca atcaaaatt tcgtcaatt atgatcctg caacttatt ttttctcga aattcgggt ccagtttta atcagtttc cgcatttac gtgtcaatg atccgaatg ataagaagg gacagatta tttttccct tgcaacatt tgcaaacga tactttcgt tggcgaaaa aactgctca cgaaagata tgttgagtt ttttggatg agcaaagtt tggaggttt atacgacgg ggaagattg ttatttttc cgtataaaa ccttctcgg agacgatta atttggtta tggtgactt tgatgaggg agttgcttc agccgcaga agagtctcc cggttgcgt tcacgaagg atgtgcaag aattcgtta aagggttgt acttaaacg gccagacgg aacttcaga gtcgcaacc acgacgcta gtgagaaag aagaccgtg aaagtaatt ttgccctcg ttaataaat tcaaagtta acgtgatgg caaacgccg cctgtgttg atcttctag tctgggcaa taggggacg aaacaggtg ccgatagtc tttgtggcc gtcttaccg aaaaaatca actcaaatc acggctaca aactcgtct tagcccaca tgcccctcg ttatggtgt gtcttgaag gcttgggca ctttagccc aaaaattcc ccaacattg cctccactt ccatttact gtcttcgag atttcctca tcactcctt cacccatcc tttccaaac tccaccgac aagcgaacg aaaaaggaa acaaggaaa atttaaaaa tcattgccc acgttaacg gccttaagg ccgaaacca ctaaaccac acaacccaa ctcaatccg catttgaga aactacgag acgctgcta ttgaaaatc tatgttatg ctcttgagg ctgcacaca ctactgatg ccgactgtg tgagggcgc tagtggcgt ccgtttcaa atcggttgt ttcgcgctg aaaaaagtg acagtgaat cgactctca tctctacta atattgtta taatgttgg gtcatgtgg ccgtttcga tgaaaggca ccccaaaga caacggata cgattttgc agtttttcc aacagagat tttgtcgag tgccttcgg cacttgata cacttattg ttgaccagt tgggttgta tgattatcg cacagtgtg aaccgcatg tttacgaag ttgtaatga agatgttta cgatgatta cggtggaga gtagcaatg cggcgtatg ggaatccca ccaatgaga aggcacagt ataggcacg tttctcgcc ctgtgatca acgcattgg ccaacatcg ccgccaata tacaaagcg aatcccaaa tgaacgaac tattaggcc gcctgctcg aactcttcg tgcaattgg gcctcgagg gcaaacgcg caagcgaaa aatccccag gcgccctaa aagcgtcaa gttcagccg gtaatttag gagttttaa tccctgtaa ttgccgttc ttctacgcc gactaccac ccatcaaaa atcccaaac ctcggattc ataaattgt tccgcgatt tttggctct attgtgtta ttatgggat ttacagcgt ctgattcgg gattatggc cgaaagaat ggtacgaag gagtgaaag aaatcgcag ttaaaagcc cagcgttag tttcaccaa cttccagtc gctcggaaa tgcgcgaat tgcaataca cgagcgccg ttcgaaacg atagtgttt caataaccg aactacaag aattgaaga atcaaattc tagaattgt tgaaacaac cggcagatg tcaccgcct atgtgaatc gcttgactt ttgcccaat gtacgttct tattgagcg tttattggg tggagattt tgcgaattc agaattcac ctgagccga gcttggtgc cgattatta ccgaatact tgtcggatt cagcgttac agaaagaca agtccggga tgtgggttt gtgtgtcgg ctgttggcg agcgcgttt ttgaaaaat tcctcgaag ttatgaaaa ataagccga agtatgagg ccagggagg cggagcttg aaggccacg cccagtttc ttctcgtac atttcaacg atccgcaca aacagattc gtcacgttt cggataagt ttctggtca gtcttttcg atcgctttc cgcacttgt tgtggagtc ggaaggtgt tgtggacaa tgcttgaca taatgcaag tgttgtcga attccttgc atttggatc cgaatcagg aaacgccca ctttgcgaa taccacgaa cgccatact ccatacagt tgatggaca cgttagagg cacgcgaga ctaaagtta agcattttg ctgcgaatt ccgagcgaa ttatcaagg aggcgttga agtgggcac cgcattgga cgaggtcgc acatacaag agtatataa taaatcaag gacaggggg cggggcttt ggaaccaca ctggtctgt cattggcgc ttgaaacta tactacagt tcggacctt tgaatatgt caagtgcac cgatgagtg tctcaacgt tggagaaga gaccgaagt gtgttaaaa gtgattcgt cgaaattga tggtttcga cgtctttaa ggtgcaaat atattggag aggtaattg ggttatcgg ccgtttatg gtaccgaag gtaaagaaa aattgattg aatttataa tgaaacggg tgtggacag cgtgtcagg aacggtccg attctgaac atcgagatg ccttgtggc aagctacgg cactcttga tttccacga cagagtttc accggaatt tgttgcatt gcattgcgt ggtcgcaag tcgagcttt tcacagtgg aagcaatgc gaacagccg tcgaatgct ggcaatggc tcataacat cacgtccag aactcgaga tccgtttcc tccaggaaa tggtctcgg cctggaact gcaccgtcc agaaacgca tgggtcttt tttcgacca gcgaaccca tgacaagcc ccctagcgg cgtacgaag gcgcccgac ttgaaccga accccccat tcgtaaaac cgcacggaa tttgggtcc agttcattt gcgaattga tcgacaacg tcaaataca gcagttacg agaaagtcg aaatgttgg caagcttga tccatcgct cgttggcaa tgtgcgtgg gggccgacc ccccatgcc aaacccgct ccgtttcag cggtcggtg tccgcttca agctactaa cttgcggct tatcgctac tccaaggcg atattttgc cgaaaagtc tggcgaaaa atgtgctca gagagcgca tttattgca gctgtttgg actatttct gcaaaccgg tcatgtgtc cgtcgcaaa caccgaccg aattgcgtg aggatatca cgacacttg tgcgatttt ggcaaacgt tacactctg ataagaagt atttgaaag ccagcgatg ttggcgatt tggacattg gtcagaatg caccaatga tggtcgaga ataacgaac tgacgaaac ctaacgact tcaaccgac caacttcgg gttggatca acaccgtac cactgtcca gcagtaccg ccaccttaa gcaaacgct cggcgaagt caaaacgcg tcccgatgg ccgacaact tcgtcaaat gttatttga aaaaacgca acctaatta tggacttgt taacggtcg agattgaat ttttgatcg tttggctca acccgactt cgcgccagg aacagcaaa tcccaggcg aggaaaaca tagcatcat ggagggcta agaccatca ccgagaaac aatggcaag actacacaa ggcttgctt gggatattt cgccggttt tggccactt atctgccct ctagattca aaacaaacg aagctataa tgagcgaaa tcaggcatc aagtgcaac aaaacccag tcagtgtgt cgcacgttc ccgaagcgt tgcaatacc tggccacca ctaacgcca tcttgagcg acagcacta aactagtgc acatgctga catgggccc gggtctccc ctatcactg ccttggcct acttcagtc gacaattcc ccccgcatc ctattacag cacagtatg ctgtgaaag tgttgagtt cgtatccgg ccgatgcgg ttttgtttt atattccgc agttggtgc aagcgttac gccacgata cgatggggt atgtgactg aatttatta aatatgtgg cgaaaaaat cgcaaattg tggcacatc agttgattt ggaatatga aaactaata tgtatttgg atgaggata tgcagcata aggatgttg tcttgtttg atgttttgg atgcgttgt gtaatagta ttttggctg agctgtcgg ggccagcga aacagtttt atgaacgag aatttgatt tctttgata agattacga gcatttcgg gggagatac ggccctatc cgaaggggc ctgagcgca ggagggcct gtctggaag cgttgagaa aaattaagg tccaacctg gttgctact tgcccagca accccgagg caatggtgg tcgacatcg actacaaca gtggcgtac cgatgcaaa gcgccgcca aagccccat acttagccc gtttcaaag tgtgccgat gcggaataa acgaactcg aaaacatcg caatggctg tatccgtta acgaaaacc atgaacaaa acttcggtc cggaaatgt ggcaagcgg ccatattta aagtcggcg atgacgtcc ggcaagaca tgttggcac tgcaagtga tcggaatat ttaagaata ttttccaaa cggtcggtt tggatttat atttgtttc cctatcggg tcgtagcca ccgcacctg ggtgtgggg tgattgaat gcgtaccaa acgccaagt cacgtgacc agttagggc ggcagaccg acatcggac tgtatgaat attttttga aaaaatacg gggaggaga gttcaaagg agtttcaaa acgcgcgcc gaaatttca tcatatcca tggccgcct acagcgttg ttggctttc ttttacaaa tcaaagacc gtcacaatg gcaatatta tgttggaca cggacggtc atataatcc acattgatt ttggtttca tgttcgagt catcgccag gcggtaact tgggttttg agcccgata ttaagctca ccgatgaga tggttatga taatggggg gcaaaatgg aagcagccc cctttaaat ggttttcgg acctgtgtg tccaggctt atttggccg tcaggcctt atcaggagt ccatcattt cgctggtct cgctcatgt tggacacgg ggctgccct gctttcggg ggcagacga tcaagttgt taagggggc gcttccatc cgggggcca ccgataagg aagcggcca cttacatgc tgcaaatca tacgtaaca gtttcctga atttcagga caaggacct atgacatga tccaatatt atcagaatc agattcctt attaactgg gagtagtgg tttttgttc taggtttta ggaat
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