| General information | |||
| TC002957 | 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_007418.3: 5820132..5827882 | |
| 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 | TC002957 | ||
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_07970 | 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 |
||
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.
Lethality 11 days after pupal injection: 0.0% (includes death as pupa, adult)
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.
Lethalities 11 days after larval injection: % (includes death as larva, prepupa, pupa)
Lethalities 22 days after larval injection: % (includes death as larva, prepupa, pupa, adult)
- >TC002957 RA
MFPTFISIL DIQSWWEVP SIAHFCSLF RTAFNLLDF DIEDLEEAL LTDGTEETS WLQELIVRL LSGCLPNNE ISTFNYQMF LRRLFRQKC QEHDQYNPF NTDVDFQLL PLRTKVDIL HALCDFRLD ADDVLDQLK NLEADSLRV EPLGYDSNE SAYWYFYGT RLYREDFKT KNNKKKSVW QVICFTEDD WFQLNKKFK KSTAKCERE LHRTLSENF LPELPRLFR EKERLARKR LLENQPRRT SRRLRKENT TEKEEVTCE SPKTNPLEE EERQRKRDL EIKRKEAYE RSNRLKEQS EDSQSEDSE VRYEMGGTK TRQKKSKGR QRKLSTSSS NSKDASSAP KSSKRKKTG EETSKSSTS SATTTVGRQ TNNSLAAAT GQIVIQPVT NSHKKKLKT SQVFRQTDE DLITGMHKI LDYVKNHED AWPFADPVD EDYAPNYYT VIRKPMDLQ RMEERLDAG YYKNFAKFR DDFQLIVDN CRLYNGVEN EYTEMVDNL LKVFERATE KFLDQISSS DDEIAVEFP NTDLDAKDT KKNNKPESK SSKRKPTLA AKTDKKREH SPSEESLGK RSLSCDSLI EEDEKISSN KKDKNTKKT NKVAKKDKN DTKKNTRDK KNSNSKHKK EKFSSPLRS RSPSRSVSR DSFRRSYSS SPPPSSVPS SPDNRFDFF PKKHKSFED KDSDTHSGP KGKQKQKQD DTNTCDTSA KKQNDKPVT SPSKPKDKH NKLRETIEK LKAKSEISN KLNDDEFRF DLDTFKENK EKSKKDDKS RKKKDKVTK PHLKKDLDF EDSNSTMES LHTTDNKKK KTNTKNCKN ASIDALTIA TEQTLKDIN KWLDDTPKY SEFSSASNS PCYPPLDDF DIVNKIDND PRKKVEKFN SKKDGIGKD IKRRTFSRD PAKFLKRRE VQRTIDRLQ PGKSKGNLI SSVQSSKQT DELFPLGPL SKNKDTKNS LIVKTDDNA PKLSLGSVL DSFGRHKFV DDTKQDESE NATKKDEKF SISTEIKEE TQIKPKLED VKKESPVRE ESSMMSGAT PNLSAWFKA FGAPKIQSS QKKSEKSEG KESEKTDDS KVDPKSKEE AKVPVPEHS PSTDSPVAT HGQPVPRQR KVSTGSSIS ERSSFSQDM DSPRVGIDE RLGAYPAPY PSPMHRSPS GASPVMASP RPDVSPKAA PYPPFNGQI RVGFYQDTV SNKSSPDKS CSPRDNPQS PYAQCSEHV YTPNTTQNM PYTYPNSPY YSHTPNYSN TNPTPPYNP DSVNPPSYY DTNKSLTDQ YQAKNIQNY VNSPNNVQS PSPHPPIET FRQEDQNQN KVNEKMVQS AMFPVKKRA YNESDSAQM SHRYEEVNR NIQPMEGQN TQNVDTMHM NAREESMKH YIHQNIPDP KPLTVQTNV SPQQQLASP IINYSTPSN PYNNYSNES NYPLNILSR SLASTPGIE RKLDIGKYT NMGYSGPEV NFPRGMQQY SRSELSYAR PPSNDQQPV SNQGQQPSE VLHYSNASP HMGYKNVDM NAATSNPQQ AHTNYTNTS VHSNPNRLG QPLDVQSSY KSNEVSIPR PTYNTHIDM DQPLGLRGN IANLSHIVD RYNNDERML TGLQPAASQ YYGDKGLGG PHMFNKPIC TSSSGLPIF NQPNMIQSY NQNMQVPTS SMYNRQMTE LQNPMSGEA KNANVAQQG SEKKSKKRK SAKTAQAAS ASEPPAQAN NQGFQSYAG LKSTSPMEP SAISLKTAS VVPGSAFNF GPSPSALGI GSGLYGDKD AYPNFLEDF RAAPNYYIA AATAAHHRS TPDAPDKQT RVAAHQSSA RQAPSYPFI GARQATPPS YPLSSHFMP SAQAPLMDA SSPLYQHYL QAGVLNQGL LGPPGPYPP GYHPALSMR QPYDSMTRP PWL
- >TC002957 RA
atgtttcca actttcatt agcatccta gatattcaa tcatggtgg gaggttcca agcattgca catttttgt tcattgttt cgaacagca tttaacctt ttggatttt gatatcgag gatttggag gaggctctc ttgactgat gggacggag gaaacctct tggcttcaa gaactgatc gtgaggctt ctcagtggg tgcttgccg aacaatgaa atctcaacg ttcaattat cagatgttc ctgagacgt ctctttcgc cagaaatgt caggaacat gatcaatac aacccattt aacacagac gttgatttc caattgtta cctttgaga acaaaagtg gacatttta catgcactc tgtgatttt cgtttagat gctgatgat gtgcttgac cagctgaaa aatttagaa gcagacagc ttgagggtt gagcccctg ggttatgac agtaacgaa tctgcttac tggtatttt tacggtact agactgtac agagaggac tttaaaacc aagaataat aagaagaaa tccgtgtgg caggttatt tgtttcact gaagacgat tggtttcaa cttaacaag aaatttaaa aaatcgact gctaagtgt gaaagagag ttacatcgt actttaagt gagaacttc ttgcctgaa ttgcccaga ttgttccgg gaaaaggaa agactggcc aggaagcgt ctgctggaa aatcaaccg agacgcact tctcgacgt ttgagaaaa gaaaataca actgaaaaa gaagaagta acgtgtgaa tcgccgaaa accaatcct cttgaagag gaagaaaga caaagaaag cgcgatttg gaaatcaag agaaaagaa gcttatgag agaagtaac agacttaaa gagcagagc gaagatagc caaagtgag gactctgaa gtacggtat gagatgggc gggactaaa acgcgacaa aagaagtcc aaaggaaga caaagaaaa ttgtcaaca agttcctct aacagtaaa gatgcatca tctgcgccg aagagctca aaaaggaag aaaacaggt gaggagact tcaaaatcc tcgacttct tccgcaact acaacagtt ggtcgacaa accaacaat tcgctggcg gcagctact ggacaaatt gttatccaa cctgttacc aactcacac aagaaaaaa ctaaaaact tctcaagtg tttcgtcaa accgacgaa gatctaata acaggaatg cacaagatc ttagattac gtaaaaaat catgaagac gcatggccc tttgccgat cctgttgat gaggattac gcacccaat tattacaca gttattcgg aaaccaatg gatttgcaa agaatggaa gagaggctg gatgcaggt tattacaag aactttgca aaattcaga gacgatttc cagttaatc gtggacaat tgtcgactt tataacggt gttgaaaat gaatatacg gaaatggtc gataatttg ttgaaagta tttgaaaga gcaactgag aaatttttg gaccaaatc tcatcatcc gacgacgaa atcgccgtt gaattccct aatactgat ctggacgca aaagatacc aaaaagaac aacaaacca gaatctaaa tcatcaaag cgtaaaccg actcttgca gctaaaaca gataaaaag agggagcat tcgccctct gaagaatct ctaggtaaa aggagtctt tcctgtgat tcgcttatt gaggaagat gaaaagata tcgtcgaat aaaaaagac aaaaacact aagaaaaca aataaagta gctaaaaag gataaaaac gacactaag aaaaacaca cgagataag aagaattct aatagtaaa cacaagaaa gagaaattt tcaagtccc ttgagaagt agaagtcct agcaggagc gttagtcga gattccttt aggcgaagt tatagcagt agtcctccg ccctcaagt gtcccttcc agtcccgat aacaggttc gattttttc ccaaaaaaa cataagtct tttgaagat aaggactcg gacacccat tcagggcct aaaggtaaa cagaagcag aagcaagac gacacaaac acatgtgat acttcagca aagaaacaa aatgacaaa ccggtcaca tcgccatca aaaccgaaa gataaacat aacaagctt cgagaaact atagaaaaa ttaaaagcc aaaagtgaa atttcgaat aaattgaac gacgacgag ttccgtttc gacttggat acctttaaa gaaaacaag gaaaagtcc aagaaagac gacaaaagt cgaaagaag aaggataaa gtgactaag ccacacttg aagaaagac ctcgatttt gaagatagt aatagcaca atggagagt ttgcatact actgacaat aagaagaag aaaacaaac acgaaaaat tgtaaaaat gcgtcgatt gatgctttg acaattgct acagaacag acgctgaaa gacatcaac aagtggtta gacgacaca ccaaaatat tcggaattt agttcggcg agtaattct ccgtgttat cctcctttg gatgacttc gatattgtc aataagatt gacaacgat cctcggaag aaagtcgaa aaattcaat tcgaaaaag gacggaatt ggtaaagat ataaagaga cgaacattt agtagagat ccggcgaaa tttctcaaa cgccgggaa gtgcaaagg accatagat cgtctacag cctggaaaa agcaaaggc aatttgatt tcgagcgtt cagagttcg aaacaaaca gacgaactg tttcctcta ggtccgcta tcaaagaat aaagacaca aaaaattct ttgatcgtt aaaactgac gataacgca ccaaagttg agtttaggc tcggttttg gacagtttc ggacggcac aagtttgtc gacgacacg aaacaggac gaatccgaa aatgcgacc aaaaaagac gaaaagttt agcatcagt acggaaatt aaagaggaa actcaaatt aaacccaaa ctggaagat gtaaagaaa gaatctccc gtgcgagaa gaatctagt atgatgagt ggagcaact ccaaattta agtgcctgg tttaaagct ttcggtgcc ccgaaaatt caatcatcc caaaagaaa agtgaaaag tcggaagga aaagaaagc gagaaaact gacgatagt aaagttgat cccaaatcg aaggaggaa gcaaaagtt ccagtccct gaacattca ccgagcact gacagcccc gtggccact cacggccaa cccgtgcca agacaaagg aaagtgagt actggaagt agcatatcg gagcgttcg tcattcagc caagacatg gactcgccc agagtggga atcgacgaa agacttggg gcctacccg gccccctac ccatccccc atgcaccgt tcgccctct ggggcctcg ccagtgatg gcctcgccc cgtccggac gtctccccc aaagcagcc ccatatcct cccttcaac ggacaaatc agagtaggg ttctaccaa gacacggtc tcgaacaaa agcagccct gacaagtcg tgcagcccc cgagacaat ccccagtcg ccctacgcc caatgctct gaacacgtc tacacccct aataccacc caaaacatg ccttatacg tatcccaac tcgccctat tactcacac acaccgaat tatagcaac acgaatcct acgcctccc tataatccg gatagtgtg aaccctcca tcttactac gacacgaat aagtcacta acggatcag taccaagcc aagaatata caaaattat gttaattcg cccaataat gtgcaatca ccgtcgcca catcctcca atagagacg ttccgacaa gaggatcag aaccagaac aaagtgaac gagaagatg gtgcaaagt gcgatgttt ccagtcaag aaacgggca tacaatgag tcagactcg gcccagatg agtcacagg tacgaggaa gtcaaccga aatatccaa cccatggag ggccaaaac acccagaat gttgacacc atgcatatg aatgcgcgt gaggagtcc atgaaacac tacatccat caaaatatc ccagaccct aaaccacta acagtccag accaatgtt tcgccccaa caacaactc gcctccccg attataaac tactctacg ccgtcaaat ccgtacaat aactacagt aacgagtct aattacccc ctaaatata ctatcgcga agtctggcc tcgacccca ggcattgag cgaaaactg gacattggg aaatatacc aatatgggg tactccggc cccgaggtg aacttccca aggggtatg caacagtac agtcggtca gagttgagc tatgcgaga cccccatcc aacgaccaa caaccagta tccaaccaa ggacaacaa ccatccgag gttttgcac tactcgaac gccagcccc catatgggg tacaaaaac gtcgacatg aacgcagcg acttccaac ccacaacaa gcacacacc aactataca aacacttcg gtgcattcg aaccccaat cgtttaggg cagcctcta gacgtgcag agttcgtac aagtcgaac gaagtgtcg atacccagg ccgacgtac aatactcac atcgatatg gaccaacct ttaggttta agagggaat atagcgaat ttgtcgcat atcgtcgac cggtataat aatgacgag aggatgttg accggattg caaccagca gcaagtcag tattatggt gataagggc ttaggaggg ccccatatg tttaataaa cccatctgt acgagctcg tcaggactg ccgattttt aatcagccc aatatgata cagtcatat aatcagaat atgcaagtg ccgacttct tcaatgtat aataggcag atgacggaa ttgcagaat ccgatgagt ggcgaagcg aagaatgcc aatgttgcg caacagggg agtgagaag aagtccaag aagcggaag agtgcaaaa acagcccag gccgcgtcc gcgtcagag ccgccggcg caggccaac aatcagggc ttccagtcg tacgcgggc ttgaagtcg acctccccc atggaacca agtgcaata tcgctgaag acagcgagc gtcgtcccc ggcagcgca ttcaatttc ggaccatcg ccgtcagcg ctgggcata ggctcaggc ctgtacggt gacaaagat gcatatccc aacttcctg gaggacttc cgcgccgcc cccaactac tacatcgcg gcggcgacg gcggcccac cacagatcc accccggac gcccccgac aagcaaacc cgcgtggcg gcgcaccag tcgtccgcc cggcaggcc cccagttac cccttcata ggcgcgcgg caggcgacg ccccctagt taccccctg agcagccac ttcatgccg tctgcgcag gcgcccctc atggatgcc agctcgccg ctctaccag cactatctg caagcgggg gtgctgaac cagggactt ttgggcccc ccagggccg tacccgccc gggtaccat cccgcgcta agtatgagg caaccatac gattccatg acaaggccg ccttggctc taa
- >TC002957 RA
gagtcgtaa attcgttaa ttcatggct ttttgggga gtttcgtcg agattaatg tgaattaac aacgttaca tgtattgat caccgtata ggttatgtt tccaacttt cattagcat cctagatat tcaatcatg gtgggaggt tccaagcat tgcacattt ttgttcatt gtttcgaac agcatttaa ccttttgga ttttgatat cgaggattt ggaggaggc tctcttgac tgatgggac ggaggaaac ctcttggct tcaagaact gatcgtgag gcttctcag tgggtgctt gccgaacaa tgaaatctc aacgttcaa ttatcagat gttcctgag acgtctctt tcgccagaa atgtcagga acatgatca atacaaccc atttaacac agacgttga tttccaatt gttaccttt gagaacaaa agtggacat tttacatgc actctgtga ttttcgttt agatgctga tgatgtgct tgaccagct gaaaaattt agaagcaga cagcttgag ggttgagcc cctgggtta tgacagtaa cgaatctgc ttactggta tttttacgg tactagact gtacagaga ggactttaa aaccaagaa taataagaa gaaatccgt gtggcaggt tatttgttt cactgaaga cgattggtt tcaacttaa caagaaatt taaaaaatc gactgctaa gtgtgaaag agagttaca tcgtacttt aagtgagaa cttcttgcc tgaattgcc cagattgtt ccgggaaaa ggaaagact ggccaggaa gcgtctgct ggaaaatca accgagacg cacttctcg acgtttgag aaaagaaaa tacaactga aaaagaaga agtaacgtg tgaatcgcc gaaaaccaa tcctcttga agaggaaga aagacaaag aaagcgcga tttggaaat caagagaaa agaagctta tgagagaag taacagact taaagagca gagcgaaga tagccaaag tgaggactc tgaagtacg gtatgagat gggcgggac taaaacgcg acaaaagaa gtccaaagg aagacaaag aaaattgtc aacaagttc ctctaacag taaagatgc atcatctgc gccgaagag ctcaaaaag gaagaaaac aggtgagga gacttcaaa atcctcgac ttcttccgc aactacaac agttggtcg acaaaccaa caattcgct ggcggcagc tactggaca aattgttat ccaacctgt taccaactc acacaagaa aaaactaaa aacttctca agtgtttcg tcaaaccga cgaagatct aataacagg aatgcacaa gatcttaga ttacgtaaa aaatcatga agacgcatg gccctttgc cgatcctgt tgatgagga ttacgcacc caattatta cacagttat tcggaaacc aatggattt gcaaagaat ggaagagag gctggatgc aggttatta caagaactt tgcaaaatt cagagacga tttccagtt aatcgtgga caattgtcg actttataa cggtgttga aaatgaata tacggaaat ggtcgataa tttgttgaa agtatttga aagagcaac tgagaaatt tttggacca aatctcatc atccgacga cgaaatcgc cgttgaatt ccctaatac tgatctgga cgcaaaaga taccaaaaa gaacaacaa accagaatc taaatcatc aaagcgtaa accgactct tgcagctaa aacagataa aaagaggga gcattcgcc ctctgaaga atctctagg taaaaggag tctttcctg tgattcgct tattgagga agatgaaaa gatatcgtc gaataaaaa agacaaaaa cactaagaa aacaaataa agtagctaa aaaggataa aaacgacac taagaaaaa cacacgaga taagaagaa ttctaatag taaacacaa gaaagagaa attttcaag tcccttgag aagtagaag tcctagcag gagcgttag tcgagattc ctttaggcg aagttatag cagtagtcc tccgccctc aagtgtccc ttccagtcc cgataacag gttcgattt tttcccaaa aaaacataa gtcttttga agataagga ctcggacac ccattcagg gcctaaagg taaacagaa gcagaagca agacgacac aaacacatg tgatacttc agcaaagaa acaaaatga caaaccggt cacatcgcc atcaaaacc gaaagataa acataacaa gcttcgaga aactataga aaaattaaa agccaaaag tgaaatttc gaataaatt gaacgacga cgagttccg tttcgactt ggatacctt taaagaaaa caaggaaaa gtccaagaa agacgacaa aagtcgaaa gaagaagga taaagtgac taagccaca cttgaagaa agacctcga ttttgaaga tagtaatag cacaatgga gagtttgca tactactga caataagaa gaagaaaac aaacacgaa aaattgtaa aaatgcgtc gattgatgc tttgacaat tgctacaga acagacgct gaaagacat caacaagtg gttagacga cacaccaaa atattcgga atttagttc ggcgagtaa ttctccgtg ttatcctcc tttggatga cttcgatat tgtcaataa gattgacaa cgatcctcg gaagaaagt cgaaaaatt caattcgaa aaaggacgg aattggtaa agatataaa gagacgaac atttagtag agatccggc gaaatttct caaacgccg ggaagtgca aaggaccat agatcgtct acagcctgg aaaaagcaa aggcaattt gatttcgag cgttcagag ttcgaaaca aacagacga actgtttcc tctaggtcc gctatcaaa gaataaaga cacaaaaaa ttctttgat cgttaaaac tgacgataa cgcaccaaa gttgagttt aggctcggt tttggacag tttcggacg gcacaagtt tgtcgacga cacgaaaca ggacgaatc cgaaaatgc gaccaaaaa agacgaaaa gtttagcat cagtacgga aattaaaga ggaaactca aattaaacc caaactgga agatgtaaa gaaagaatc tcccgtgcg agaagaatc tagtatgat gagtggagc aactccaaa tttaagtgc ctggtttaa agctttcgg tgccccgaa aattcaatc atcccaaaa gaaaagtga aaagtcgga aggaaaaga aagcgagaa aactgacga tagtaaagt tgatcccaa atcgaagga ggaagcaaa agttccagt ccctgaaca ttcaccgag cactgacag ccccgtggc cactcacgg ccaacccgt gccaagaca aaggaaagt gagtactgg aagtagcat atcggagcg ttcgtcatt cagccaaga catggactc gcccagagt gggaatcga cgaaagact tggggccta cccggcccc ctacccatc ccccatgca ccgttcgcc ctctggggc ctcgccagt gatggcctc gccccgtcc ggacgtctc ccccaaagc agccccata tcctccctt caacggaca aatcagagt agggttcta ccaagacac ggtctcgaa caaaagcag ccctgacaa gtcgtgcag cccccgaga caatcccca gtcgcccta cgcccaatg ctctgaaca cgtctacac ccctaatac cacccaaaa catgcctta tacgtatcc caactcgcc ctattactc acacacacc gaattatag caacacgaa tcctacgcc tccctataa tccggatag tgtgaaccc tccatctta ctacgacac gaataagtc actaacgga tcagtacca agccaagaa tatacaaaa ttatgttaa ttcgcccaa taatgtgca atcaccgtc gccacatcc tccaataga gacgttccg acaagagga tcagaacca gaacaaagt gaacgagaa gatggtgca aagtgcgat gtttccagt caagaaacg ggcatacaa tgagtcaga ctcggccca gatgagtca caggtacga ggaagtcaa ccgaaatat ccaacccat ggagggcca aaacaccca gaatgttga caccatgca tatgaatgc gcgtgagga gtccatgaa acactacat ccatcaaaa tatcccaga ccctaaacc actaacagt ccagaccaa tgtttcgcc ccaacaaca actcgcctc cccgattat aaactactc tacgccgtc aaatccgta caataacta cagtaacga gtctaatta ccccctaaa tatactatc gcgaagtct ggcctcgac cccaggcat tgagcgaaa actggacat tgggaaata taccaatat ggggtactc cggccccga ggtgaactt cccaagggg tatgcaaca gtacagtcg gtcagagtt gagctatgc gagaccccc atccaacga ccaacaacc agtatccaa ccaaggaca acaaccatc cgaggtttt gcactactc gaacgccag cccccatat ggggtacaa aaacgtcga catgaacgc agcgacttc caacccaca acaagcaca caccaacta tacaaacac ttcggtgca ttcgaaccc caatcgttt agggcagcc tctagacgt gcagagttc gtacaagtc gaacgaagt gtcgatacc caggccgac gtacaatac tcacatcga tatggacca acctttagg tttaagagg gaatatagc gaatttgtc gcatatcgt cgaccggta taataatga cgagaggat gttgaccgg attgcaacc agcagcaag tcagtatta tggtgataa gggcttagg agggcccca tatgtttaa taaacccat ctgtacgag ctcgtcagg actgccgat ttttaatca gcccaatat gatacagtc atataatca gaatatgca agtgccgac ttcttcaat gtataatag gcagatgac ggaattgca gaatccgat gagtggcga agcgaagaa tgccaatgt tgcgcaaca ggggagtga gaagaagtc caagaagcg gaagagtgc aaaaacagc ccaggccgc gtccgcgtc agagccgcc ggcgcaggc caacaatca gggcttcca gtcgtacgc gggcttgaa gtcgacctc ccccatgga accaagtgc aatatcgct gaagacagc gagcgtcgt ccccggcag cgcattcaa tttcggacc atcgccgtc agcgctggg cataggctc aggcctgta cggtgacaa agatgcata tcccaactt cctggagga cttccgcgc cgcccccaa ctactacat cgcggcggc gacggcggc ccaccacag atccacccc ggacgcccc cgacaagca aacccgcgt ggcggcgca ccagtcgtc cgcccggca ggcccccag ttacccctt cataggcgc gcggcaggc gacgccccc tagttaccc cctgagcag ccacttcat gccgtctgc gcaggcgcc cctcatgga tgccagctc gccgctcta ccagcacta tctgcaagc gggggtgct gaaccaggg acttttggg ccccccagg gccgtaccc gcccgggta ccatcccgc gctaagtat gaggcaacc atacgattc catgacaag gccgccttg gctctaatc gacggtttg tttttcggg aaagttatt agtgattgt ttttatttg ccaaagaca gtttattcc gtttgttaa attgtacat taaatttta ttttggaaa ttcgacgat tttgcgatt attgtttta tttttttat agcggcgtg cggaaagct ctgtgattt actttacgt gtgatcgta aatattttt gttatgtct ttttcatgt ttatggatt cactgtcaa tagtttttc gtaactgtt agcatcgga ccattattt tacctcatg ttttatatc atagtatat cgaactttt gtagtgctt acaatagtt ctgtgttgt ttattttaa aatgaaatt gttgtatac tgggtaact ctgaccaga agttgtctt cgattaaca tgttgctat acggtcggg tgtatccgc aaacattcc caaattttg tgcaaaata tctacttac ctagactaa ggttcttaa tagcagaga tatatgtca atgaattta ttattcatc acggtttga tttatgttt atacattta tgtatagta cataaatac acaacttgt gttaat
