| General information | |||
| TC002876 | 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. |
scaffold_seqid16: 3035312..3042390 | |
| 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 |
|
TC002876 | |
| General information | ||||
| iB_00461 | 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
Fertility
eggs lost during procedure: potentially due to defects in egg-shells egg-morphology or egg-size, (14 dpi)
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: 60.0% (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 delayed (still larva) - Number of animals showing the phenotype: 4 (11 dpi)
Days post injection
larva lethality late (L7) - Number of animals showing the phenotype: 1 (16 dpi)
Days post injection
pupal molt delayed (still larva) - Number of animals showing the phenotype: 3 (16 dpi)
Days post injection
remarks: later than schedule, pupal molt not fulfilled (died as L7 or prepupa) - Number of animals showing the phenotype: 4 (19 dpi)
Days post injection
Pupal morphology
remarks: later than schedule,
Adult morphology
remarks: later than schedule,
- >TC002876 RA
MSTSLSEQL KRLALPQTS LLQRDKKRP SLLFDPKEA AGLKRETVY QIGLDGLEE LITKNPVFE QFKSNIFHI TSKKFERAV QSADANKKL DKVIRKFLL QLSPYFLLN CAHKALEWL INRYNIHEF NRDDLLMLI MPYHESNIF VRVVQLMKF KDENDSWFF LKSIQKTGV HLTKQSLLN KASSDVYFL KFVAKFISL LVKQFEKPS LLTVAFNFY CTVFVGAIE YSSELNEAQ ITQMLPALL KGLNSDIPD FCAASYVIL GRLVMKTPL TDAILDKFV EKVSVSKVE TLKTESVLT LLILCQSQT HYTQLPESA CYSLIGKNW LPKVLQDLS NGNCFVTPF LKVLVKTCL YGAVKLDLE EYRAFMKTL LAAIKFDDA FMEIFLSLL LDRTKTKKP YSEHLQKWL TEIIETLER QYPDQFDKE VYKILSTTQ QGKVIKRRK SLQKILTET MNIRCKFDV LAKLYHPNV TYRKEAIKY LVNNYDSLR DREKEMIKS SFIDRLNDD SVEVVHETL NLIKNISVL KTDALKDVL TKLAYKCQS DLKTWGSVS TEIVLMLCE QSEANDWET FVAVLPYLL PNSENQLSA AQTIAESSF CRKNCLLKK INAKGEDCA QFCNSVFDS MRNNNKELV NDFIACIHK TTDRSVLSK YLIILILSI ILPENSSIE SNITIVKIF IEFIENCEI KYEKGNAFM QKHISVARA GQFHIEGFL LCLKNVIVK TLKPTVELG RVDLHEDTD FNQYFTLLA TTLLKNKSN HKKYVHVFI NYFCKTFRS KIEFLLNLV NSGKSQDLE FVTESVSLI AKLLSSVKD SNEMSSLLE SDIFIPQLL ALLLHKSED VKQKSFRIV EILTEVKND TFGHLFREL LDQREEIVV DNDQLPLIM FNYLSPSAV KDTKVSGNA STLKTLLIK TSCGKYPVY LKSKMLELL SHINTIEIF EETSHDCLH LLKTQSKNL DEVKSSIVV RNLQRCQPT IAGNIAVGT SIWTLIETC LKDSDFTLT VDSKPEFAT VLLLKQFDR EFFAQLDAK VETKLLDLI VETATLTKN PDVLPAVRH TIKVIELDA KLILSHFVK MRDVQSPKL DPNKLKRRI SVVPTVDIL DTLDWRKGL TALEFIQDK KKLTNIEVL FPVLFEILK KCLDFDEQA AVEYPKQLI LLSILYLGE KIDRDSIGE TTFNMELIV QCIRASQNP QTHYCALLV LAFSAELFP SQVLHHIMA IFTFMGSSV LRHEDAYSF QIITRIIDT VIPILVKDN NPETIARVL RVFVDALLD VPEHRRMPI YGHLLTKLQ VKDNLYIFL LLIFESHVN HSHRDKNKE DDMKRLEIA ASVCREFSP EIVIQNCIN LMKYLKELP DEVDSDNTS SPFDIHYKT PKQFRHYKY TLIIFTARL LSSKEFVFQ VASLKNDEL LDLQQLYKD MIINILTYI YRISKIADK NANTQHAKY WKVILHLSY DILDSVNAL LTPQMFLLV VKGLMLHNF STIRRRSME LLNSKLQNN PSFFDDCTS EQIYSLIPP LISIIKNVE EAQSESDDE LIVQTALLS LKLLTKFLV SDYHEKLAE ILDFVTDLA KSGKVQNNV LASVLLCLA ELCTNLRAH AIPSLPNFM PAILRILKQ QKSQESSSL LLLSSVTTV QKIVETLPL FLSPYLEKL LYELSLLAS NVAEEERLQ PVVNKLSNL KQQIGTSIP PRVLIPVLG QSYDRLITK QAFSAVGFL LDTLAENLN HLNGSEIGA NLPELTSFF LNALQFRTD QDTTFEQAN EVEAQIVKA LTKLILKLS ESTFKPLYY KLFDWAARH EQKTERLIT FYALSSGIA EALKSLFVL FAGHFLNNA AQILDSCNV TKTDALYFD DEKKNVLLL ENVLKTLHS VFLYDSHKF VNKDRFEVL MQPLVDQLE NTLGGVGGL ERRNGELVT PVIVQFAVA TADDSLWKQ LNYQILLKM KHNSPEIRL ISLNCLKEM VTKLGVDYL PLLPETIPV LAELLEDEE ESVEKACRK AVQEMEKIL GEPIEKYFK M
- >TC002876 RA
atgagcacc tcactgagc gagcaactc aagcgcctg gctttgccc caaacctcg ctgttacaa agggacaag aaacgccca tctttgctc ttcgaccct aaggaagcg gctggtttg aaacgtgag accgtttac caaatcggt cttgacggt ttagaagaa ttgatcacg aaaaaccca gtctttgag cagtttaaa agcaacatt tttcacata acttcaaaa aaatttgag cgcgcggtg caatccgct gacgcaaac aaaaagttg gataaagtg attaggaaa tttctgctg caattgtcg ccttatttt ttgctgaac tgtgcacat aaggcgctt gaatggctc ataaacaga tacaacatt catgaattt aacagagac gatttgctt atgcttata atgccgtat cacgagtcg aatattttc gttagagtg gtgcagttg atgaaattc aaggatgag aatgacagt tggttcttt ttgaagtcg attcagaaa actggggtt catctcact aaacaaagc ctactaaac aaggcatca agtgatgtt tatttccta aaatttgtg gccaaattc atctctctg ctcgtgaag caatttgaa aaaccctcg ttactcacc gtcgctttc aacttttat tgcaccgtt ttcgtgggg gctattgaa tattcctcc gagttaaat gaagctcag atcactcag atgctacca gcgctcctc aaaggccta aattcggac attccggat ttttgcgcc gcttcgtac gttattctg ggacgactt gtaatgaaa acgccgcta actgatgca attttggac aaattcgtc gaaaaagta tccgtgtcg aaagttgaa actttgaaa actgaatca gttttgact cttttgatt ctttgccaa tctcagact cattacact cagttaccg gaatcggct tgctatagt ttgattggg aaaaattgg ttaccgaaa gtcttgcaa gatttgagc aacggaaat tgtttcgtt accccattt ttgaaggtt ttggtcaag acgtgcttg tatggggct gtcaaactg gacctggag gagtatcgc gcttttatg aagactctt ctagcggcg attaaattc gacgacgct ttcatggaa atattttta agtcttctt ctcgaccgt accaagacc aagaaaccc tactcggaa cacctccaa aaatggcta acggaaata atcgaaacg ctcgaacgc caatacccc gaccaattc gataaagaa gtctacaaa atcctctcg acaacgcaa caaggcaaa gtgatcaag cgacgaaaa tcgcttcaa aaaatcctc accgaaaca atgaacatt cggtgcaag ttcgacgtt ttggccaag ttatatcac cccaacgtc acataccga aaagaagcc atcaaatat cttgtcaat aattatgac tctctccgc gaccgtgaa aaagaaatg atcaaaagc tcgttcatt gatcgcctg aacgacgac agcgtggaa gtcgttcat gaaacacta aacctgatc aaaaatata tctgttctg aagacagac gcgctgaag gacgttttg accaaactg gcctacaag tgccagagc gatttgaag acttggggg tccgtctcg accgaaatt gtgctcatg ttgtgcgag caaagcgag gcaaacgac tgggaaacg ttcgtagcc gtgcttccc tacttatta ccaaactct gaaaatcag ttaagtgca gcacaaaca attgccgaa agttcgttc tgtcgtaaa aattgcttg ctgaagaag attaacgcc aaaggtgag gattgtgcc cagttttgt aacagcgtt tttgactca atgcggaac aacaacaaa gaactggtc aacgacttt atcgcctgt attcacaaa acgacagat cgttccgta ctcagcaaa tatctgata attcttatt ctgagcatc atcctacct gaaaatagc tcaattgag agcaacatc acaatcgtc aaaatcttt atcgagttt atcgaaaat tgtgagatt aaatatgaa aaagggaac gcttttatg caaaaacac atcagtgtg gctcgcgcc ggccagttc cacatcgaa gggtttcta ctgtgtttg aaaaacgta attgtgaag acacttaaa cccacggtt gagttgggc cgagttgac ttacatgag gacactgat tttaatcaa tactttact ttacttgcc accacttta ctcaaaaac aagtcaaac cacaaaaag tacgtccac gtttttatt aactacttc tgcaaaacg ttccgaagc aaaatagag tttttatta aacttggtc aacagtggg aagagccag gatttagag ttcgtgacc gaaagtgtg tcactaatt gcgaaattg ctgtccagt gtcaaggac agcaacgaa atgagttcg cttctcgag agtgatatt tttatcccc caacttctg gccttgttg ttacacaaa tcggaagac gtcaagcaa aaaagtttc cgaattgtt gaaatctta accgaagtg aagaacgat acttttggc cacctcttt agggagttg ttggaccaa agggaggag attgttgtt gacaatgac cagttgcca ctcatcatg tttaactac ttgtcacct tctgcagtc aaagacacg aaagtttct ggcaatgca tcaacgctt aaaacacta ttaattaaa acaagctgt ggcaaatac ccggtctat ctgaaatcg aaaatgttg gagctccta tcccacatt aacacgatc gaaatcttc gaggaaacg tcgcacgat tgcttgcac ttgctcaaa acccagagc aagaatttg gacgaagtc aagtcatcg atagtcgtc cgaaacttg caacgttgc cagcccaca atcgccgga aatatcgca gtggggacg tccatctgg actttgatt gagacgtgt ctcaaagac tcggacttc actctcact gttgactcc aaaccggag tttgcaaca gtattactt ttgaagcag ttcgatcgc gagtttttc gcccaactg gacgcgaaa gtcgagacg aagctcctc gacttgatc gtcgaaacg gccactctg accaaaaac ccggacgtt ctgccggcc gttcggcac acgatcaag gtgatagaa ctggacgcg aagctaatt ctgagccat ttcgtcaaa atgcgagac gtgcagtcg cccaaactc gaccccaac aagctaaaa cggcggatc agtgttgtg cccactgtg gatattttg gacactttg gactggcgc aagggcctg acggcgctc gagtttatc caagacaag aagaaactg accaatatt gaggtcctg ttccccgtg ttgttcgag attttgaag aagtgtctc gattttgac gaacaggcg gccgtggag taccccaag cagttgatc ctcttgtcg attctctac ctgggggag aaaatcgat agggatagt ataggagag actacgttc aacatggag ttgattgtt cagtgtatc agggcgtcg caaaacccg cagacccac tattgcgcc cttttggtg ctggccttt tcggcggag ttgttccct tcgcaagtt ttgcaccac ataatggcc attttcacg tttatgggc tcgtctgtt ctgagacac gaggatgcg tacagtttt cagatcatt acgaggatt atcgatact gtgataccg attctagtc aaggataat aatcctgaa actatagcc agggttttg agggttttt gtggacgcc cttctggac gttcccgag caccggagg atgccgatt tatgggcac ttgctgacg aagttgcaa gtcaaggac aatctttac atatttttg ttgctgatt ttcgagtca cacgttaat cacagccac agggacaaa aacaaagag gacgatatg aagcggctc gaaatcgca gcgagtgtt tgccgcgag ttttcgcca gaaatcgta atccagaat tgtatcaac ttgatgaaa tacttgaag gagttaccc gacgaagtc gacagtgac aacacgagt tcgcctttt gatatccat tataaaacg ccaaagcaa ttccgacat tacaaatac actttaatt attttcacc gcgaggtta ctgtcgtcc aaagagttt gttttccaa gtggcttcg ctgaaaaac gacgaactg ctcgacttg cagcagctg tacaaagac atgattata aacatcctg acgtacatt taccgcatt tccaaaatt gcggacaag aacgcaaac acgcaacac gccaagtac tggaaggtc atcctccac ttgagttac gatatcctg gacagcgtc aacgcactg ttaaccccc cagatgttc ttgttggtg gtcaaaggc ctgatgctg cacaacttc agcacgatt cggaggcgc agcatggag ttgttgaac tccaaactg cagaacaac ccttcgttt ttcgacgac tgcacgtcg gagcaaatc tactcgtta atcccgccc ctcatttca atcattaag aacgtggaa gaggcgcag agcgagagc gacgacgag ctgatcgtg caaactgcg ctcctctcc ttgaagctt ctgaccaag ttcttagtc tcggactac cacgagaaa ctggccgag attctcgac ttcgtgacc gatttggcc aaaagtggc aaagtgcaa aacaacgtc cttgcctca gttttgctg tgcttggcc gagctttgc acgaacttg cgggcacat gccatcccc agcctcccg aacttcatg cccgcgatt ttgaggatt ttgaagcaa cagaagagt caagaatcg tcgagtttg ctcctcttg agttcggtc accactgtg caaaagatt gtcgaaacg ctcccactt ttcctcagc ccgtatctc gagaaatta ttgtatgag ctgtcgcta cttgcgagt aacgtagcc gaggaggaa cgcctccaa cccgttgtg aataaacta tcaaacttg aagcaacaa atcgggact tccatcccc ccacgggtg ttaataccg gttttgggg caaagctac gaccgcctg atcacgaaa caggcgttt agcgctgtg ggatttctg ctcgacact ttggccgag aatttgaac cacttgaac gggtccgaa atcggcgcg aatttgccc gaactgacc agctttttc ctgaatgcg ctccagttc aggacggat caagacacg acgttcgaa caggccaac gaagtggag gcgcaaatc gtcaaagct ttgactaaa ctgatcttg aaactgtcg gaaagtacg ttcaagccc ttgtattac aaactgttt gattgggcg gcgcgacac gagcagaaa actgaacga cttatcacg ttctatgcg ctgtctagc gggattgcc gaagctttg aaaagttta tttgtgctt ttcgctggc catttcctc aacaatgcg gcccagatt ctggattcg tgcaacgtc actaaaacc gacgctctc tatttcgac gatgagaag aagaatgtt ctcttgttg gagaatgtt ttgaagacg cttcattcc gtgtttctg tacgatagt cacaagttt gttaacaag gaccggttt gaggttttg atgcagccg ctggtggat cagttggaa aacacctta gggggtgtt gggggcttg gaaaggcgg aatggcgag ttggtgacg ccggtgatt gtgcagttt gcggtggca acggctgat gattcgttg tggaagcag cttaattac cagattttg ctgaaaatg aagcacaat tcgcctgag attaggctg atttcgttg aattgtttg aaggagatg gtgacgaag ctgggggtt gattatttg cccttgtta ccggagacg atcccagtg ttggccgaa ttgttggaa gatgaggag gaaagtgtg gagaaggcg tgcaggaag gcggtccaa gagatggag aagatttta ggggagcca attgaaaag tatttcaaa atgtaa
- >TC002876 RA
tcataacca cagtttttc agcgtgccc cagttgtca cgtgtaatg taatttctg gcgttgttt ttgtttaca ggcgccggt tttggataa aataaagat gagcacctc actgagcga gcaactcaa gcgcctggc tttgcccca aacctcgct gttacaaag ggacaagaa acgcccatc tttgctctt cgaccctaa ggaagcggc tggtttgaa acgtgagac cgtttacca aatcggtct tgacggttt agaagaatt gatcacgaa aaacccagt ctttgagca gtttaaaag caacatttt tcacataac ttcaaaaaa atttgagcg cgcggtgca atccgctga cgcaaacaa aaagttgga taaagtgat taggaaatt tctgctgca attgtcgcc ttatttttt gctgaactg tgcacataa ggcgcttga atggctcat aaacagata caacattca tgaatttaa cagagacga tttgcttat gcttataat gccgtatca cgagtcgaa tattttcgt tagagtggt gcagttgat gaaattcaa ggatgagaa tgacagttg gttcttttt gaagtcgat tcagaaaac tggggttca tctcactaa acaaagcct actaaacaa ggcatcaag tgatgttta tttcctaaa atttgtggc caaattcat ctctctgct cgtgaagca atttgaaaa accctcgtt actcaccgt cgctttcaa cttttattg caccgtttt cgtgggggc tattgaata ttcctccga gttaaatga agctcagat cactcagat gctaccagc gctcctcaa aggcctaaa ttcggacat tccggattt ttgcgccgc ttcgtacgt tattctggg acgacttgt aatgaaaac gccgctaac tgatgcaat tttggacaa attcgtcga aaaagtatc cgtgtcgaa agttgaaac tttgaaaac tgaatcagt tttgactct tttgattct ttgccaatc tcagactca ttacactca gttaccgga atcggcttg ctatagttt gattgggaa aaattggtt accgaaagt cttgcaaga tttgagcaa cggaaattg tttcgttac cccattttt gaaggtttt ggtcaagac gtgcttgta tggggctgt caaactgga cctggagga gtatcgcgc ttttatgaa gactcttct agcggcgat taaattcga cgacgcttt catggaaat atttttaag tcttcttct cgaccgtac caagaccaa gaaacccta ctcggaaca cctccaaaa atggctaac ggaaataat cgaaacgct cgaacgcca ataccccga ccaattcga taaagaagt ctacaaaat cctctcgac aacgcaaca aggcaaagt gatcaagcg acgaaaatc gcttcaaaa aatcctcac cgaaacaat gaacattcg gtgcaagtt cgacgtttt ggccaagtt atatcaccc caacgtcac ataccgaaa agaagccat caaatatct tgtcaataa ttatgactc tctccgcga ccgtgaaaa agaaatgat caaaagctc gttcattga tcgcctgaa cgacgacag cgtggaagt cgttcatga aacactaaa cctgatcaa aaatatatc tgttctgaa gacagacgc gctgaagga cgttttgac caaactggc ctacaagtg ccagagcga tttgaagac ttgggggtc cgtctcgac cgaaattgt gctcatgtt gtgcgagca aagcgaggc aaacgactg ggaaacgtt cgtagccgt gcttcccta cttattacc aaactctga aaatcagtt aagtgcagc acaaacaat tgccgaaag ttcgttctg tcgtaaaaa ttgcttgct gaagaagat taacgccaa aggtgagga ttgtgccca gttttgtaa cagcgtttt tgactcaat gcggaacaa caacaaaga actggtcaa cgactttat cgcctgtat tcacaaaac gacagatcg ttccgtact cagcaaata tctgataat tcttattct gagcatcat cctacctga aaatagctc aattgagag caacatcac aatcgtcaa aatctttat cgagtttat cgaaaattg tgagattaa atatgaaaa agggaacgc ttttatgca aaaacacat cagtgtggc tcgcgccgg ccagttcca catcgaagg gtttctact gtgtttgaa aaacgtaat tgtgaagac acttaaacc cacggttga gttgggccg agttgactt acatgagga cactgattt taatcaata ctttacttt acttgccac cactttact caaaaacaa gtcaaacca caaaaagta cgtccacgt ttttattaa ctacttctg caaaacgtt ccgaagcaa aatagagtt tttattaaa cttggtcaa cagtgggaa gagccagga tttagagtt cgtgaccga aagtgtgtc actaattgc gaaattgct gtccagtgt caaggacag caacgaaat gagttcgct tctcgagag tgatatttt tatccccca acttctggc cttgttgtt acacaaatc ggaagacgt caagcaaaa aagtttccg aattgttga aatcttaac cgaagtgaa gaacgatac ttttggcca cctctttag ggagttgtt ggaccaaag ggaggagat tgttgttga caatgacca gttgccact catcatgtt taactactt gtcaccttc tgcagtcaa agacacgaa agtttctgg caatgcatc aacgcttaa aacactatt aattaaaac aagctgtgg caaataccc ggtctatct gaaatcgaa aatgttgga gctcctatc ccacattaa cacgatcga aatcttcga ggaaacgtc gcacgattg cttgcactt gctcaaaac ccagagcaa gaatttgga cgaagtcaa gtcatcgat agtcgtccg aaacttgca acgttgcca gcccacaat cgccggaaa tatcgcagt ggggacgtc catctggac tttgattga gacgtgtct caaagactc ggacttcac tctcactgt tgactccaa accggagtt tgcaacagt attactttt gaagcagtt cgatcgcga gtttttcgc ccaactgga cgcgaaagt cgagacgaa gctcctcga cttgatcgt cgaaacggc cactctgac caaaaaccc ggacgttct gccggccgt tcggcacac gatcaaggt gatagaact ggacgcgaa gctaattct gagccattt cgtcaaaat gcgagacgt gcagtcgcc caaactcga ccccaacaa gctaaaacg gcggatcag tgttgtgcc cactgtgga tattttgga cactttgga ctggcgcaa gggcctgac ggcgctcga gtttatcca agacaagaa gaaactgac caatattga ggtcctgtt ccccgtgtt gttcgagat tttgaagaa gtgtctcga ttttgacga acaggcggc cgtggagta ccccaagca gttgatcct cttgtcgat tctctacct gggggagaa aatcgatag ggatagtat aggagagac tacgttcaa catggagtt gattgttca gtgtatcag ggcgtcgca aaacccgca gacccacta ttgcgccct tttggtgct ggccttttc ggcggagtt gttcccttc gcaagtttt gcaccacat aatggccat tttcacgtt tatgggctc gtctgttct gagacacga ggatgcgta cagttttca gatcattac gaggattat cgatactgt gataccgat tctagtcaa ggataataa tcctgaaac tatagccag ggttttgag ggtttttgt ggacgccct tctggacgt tcccgagca ccggaggat gccgattta tgggcactt gctgacgaa gttgcaagt caaggacaa tctttacat atttttgtt gctgatttt cgagtcaca cgttaatca cagccacag ggacaaaaa caaagagga cgatatgaa gcggctcga aatcgcagc gagtgtttg ccgcgagtt ttcgccaga aatcgtaat ccagaattg tatcaactt gatgaaata cttgaagga gttacccga cgaagtcga cagtgacaa cacgagttc gccttttga tatccatta taaaacgcc aaagcaatt ccgacatta caaatacac tttaattat tttcaccgc gaggttact gtcgtccaa agagtttgt tttccaagt ggcttcgct gaaaaacga cgaactgct cgacttgca gcagctgta caaagacat gattataaa catcctgac gtacattta ccgcatttc caaaattgc ggacaagaa cgcaaacac gcaacacgc caagtactg gaaggtcat cctccactt gagttacga tatcctgga cagcgtcaa cgcactgtt aacccccca gatgttctt gttggtggt caaaggcct gatgctgca caacttcag cacgattcg gaggcgcag catggagtt gttgaactc caaactgca gaacaaccc ttcgttttt cgacgactg cacgtcgga gcaaatcta ctcgttaat cccgcccct catttcaat cattaagaa cgtggaaga ggcgcagag cgagagcga cgacgagct gatcgtgca aactgcgct cctctcctt gaagcttct gaccaagtt cttagtctc ggactacca cgagaaact ggccgagat tctcgactt cgtgaccga tttggccaa aagtggcaa agtgcaaaa caacgtcct tgcctcagt tttgctgtg cttggccga gctttgcac gaacttgcg ggcacatgc catccccag cctcccgaa cttcatgcc cgcgatttt gaggatttt gaagcaaca gaagagtca agaatcgtc gagtttgct cctcttgag ttcggtcac cactgtgca aaagattgt cgaaacgct cccactttt cctcagccc gtatctcga gaaattatt gtatgagct gtcgctact tgcgagtaa cgtagccga ggaggaacg cctccaacc cgttgtgaa taaactatc aaacttgaa gcaacaaat cgggacttc catcccccc acgggtgtt aataccggt tttggggca aagctacga ccgcctgat cacgaaaca ggcgtttag cgctgtggg atttctgct cgacacttt ggccgagaa tttgaacca cttgaacgg gtccgaaat cggcgcgaa tttgcccga actgaccag ctttttcct gaatgcgct ccagttcag gacggatca agacacgac gttcgaaca ggccaacga agtggaggc gcaaatcgt caaagcttt gactaaact gatcttgaa actgtcgga aagtacgtt caagccctt gtattacaa actgtttga ttgggcggc gcgacacga gcagaaaac tgaacgact tatcacgtt ctatgcgct gtctagcgg gattgccga agctttgaa aagtttatt tgtgctttt cgctggcca tttcctcaa caatgcggc ccagattct ggattcgtg caacgtcac taaaaccga cgctctcta tttcgacga tgagaagaa gaatgttct cttgttgga gaatgtttt gaagacgct tcattccgt gtttctgta cgatagtca caagtttgt taacaagga ccggtttga ggttttgat gcagccgct ggtggatca gttggaaaa caccttagg gggtgttgg gggcttgga aaggcggaa tggcgagtt ggtgacgcc ggtgattgt gcagtttgc ggtggcaac ggctgatga ttcgttgtg gaagcagct taattacca gattttgct gaaaatgaa gcacaattc gcctgagat taggctgat ttcgttgaa ttgtttgaa ggagatggt gacgaagct gggggttga ttatttgcc cttgttacc ggagacgat cccagtgtt ggccgaatt gttggaaga tgaggagga aagtgtgga gaaggcgtg caggaaggc ggtccaaga gatggagaa gattttagg ggagccaat tgaaaagta tttcaaaat gtaaataga attttaatg atttaagaa ggcaataaa cggttttta attaataca ttgtttatt ttgta