| General information | |||
| TC003518 | 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: 6207472..6220748 | |
| Sequence information | |||
| Transcripts / Proteins (3) | 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 |
|
TC003518 | |
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_03568 | 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: 20.0% (includes death as pupa, adult)
eclosion not fulfilled (with pupal and adult features) - Number of animals showing the phenotype: 2 (3 dpi)
affected eggs/embryos/larvae total percentage of affected eggs/embryos/larvae on slide - Penetrance (percentage of animals/eggs on slide): less than 30%
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: 20.0% (includes death as larva, prepupa, pupa)
Lethalities 22 days after larval injection: 20.0% (includes death as larva, prepupa, pupa, adult)
remarks: later than schedule,
- >TC003518 RA
MNKVLPGLY VGNYRDSKD ANQLAKHNI THILAIHDS AKRIHSDKH YLCVMASDS PDQNLTQYF SICNDFIHA ARLREGNVL IHCLAGMSR SVTVAVAYI MSVTNLNWK EALKVVRAG RAVANPNLG FQKQLQDFE ATRLAKERR RLKERYPSL ALVNIDREQ CSLMLNNYE ELLQSRSIC EGRCALGEE CPTGICRSK NQNPAAHQP RSRRQSRST RSRSEMCRS PSGTSAAST STRPRSGPA GLRSYTGSA PPSRTSSRV DISSIGVSG NLTAPTTPQ TSPPISPKR RGTVAVPLA LKEDDFDL
- >TC003518 RB
MNKVLPGLY VGNYRDSKD ANQLAKHNI THILAIHDS AKRIHSDKH YLCVMASDS PDQNLTQYF SICNDFIHA ARLREGNVL IHCLAGMSR SVTVAVAYI MSVTNLNWK EALKVVRAG RAVANPNLG FQKQLQDFE ATRLAKERR RLKERYPSL ALVNIDREQ CSLMLNNYE ELLQSRSIC EGRCALGEE CPTGICRSK NQNPAAHQP RSRRQSRST RSRSEMCRS PSGTSAAST STRPRSGPA GLRSYTGAL YIYVVMVQL RHLGPVPA
- >TC003518 RC
MGNGMNKVL PGLYVGNYR DSKDANQLA KHNITHILA IHDSAKRIH SDKHYLCVM ASDSPDQNL TQYFSICND FIHAARLRE GNVLIHCLA GMSRSVTVA VAYIMSVTN LNWKEALKV VRAGRAVAN PNLGFQKQL QDFEATRLA KERRRLKER YPSLALVNI DREQCSLML NNYEELLQS RSICEGRCA LGEECPTGI CRSKNQNPA AHQPRSRRQ SRSTRSRSE MCRSPSGTS AASTSTRPR SGPAGLRSY TGSAPPSRT SSRVDISSI GVSGNLTAP TTPQTSPPI SPKRRGTVA VPLALKEDD FDL
- >TC003518 RA
atgaataag gtgttgcct ggcctttat gtgggaaat tacagggac tcaaaagac gctaaccaa ctagccaag cacaacata acacacatc cttgccatc catgactct gcgaaaaga atacattcc gataagcac tatctttgc gtgatggcg tcggattct ccggatcaa aacctgact caatatttt tccatatgc aacgatttc atccacgct gcgaggctc cgagaaggc aacgtttta atacattgt ctggctgga atgtctcgc agtgtgaca gtggcagtg gcgtacatc atgtctgta actaatctc aactggaag gaagcactt aaagtcgtt agagctggt cgcgcggta gccaacccc aacctgggc ttccaaaag caattacag gattttgaa gccacgcgg ttagcaaag gagcgccgc cgcctcaag gagcgctac ccaagttta gctttagta aacatagac agagaacag tgttctttg atgctcaat aactacgag gagttgctg caatctcgg tcaatttgc gaaggtcga tgcgcgttg ggcgaagaa tgtccaacg ggaatttgc agatcgaag aatcaaaac ccagctgct caccagccg cgttcacgg agacagagc agaagcaca agatcgcgt tcggagatg tgtcgaagt cctagtggc actagcgcc gctagcacc agcaccaga ccgcgctcg gggcccgca ggacttcgc agctataca ggttcagct ccgccatct cggaccagt tcccgcgta gatatttct tccattggt gtttccgga aaccttaca gctccaaca acaccgcaa acaagccct ccaatttca ccgaaacgg cgaggaacg gtcgcagtg cctctggct ttgaaagaa gacgatttt gatttatag
- >TC003518 RB
atgaataag gtgttgcct ggcctttat gtgggaaat tacagggac tcaaaagac gctaaccaa ctagccaag cacaacata acacacatc cttgccatc catgactct gcgaaaaga atacattcc gataagcac tatctttgc gtgatggcg tcggattct ccggatcaa aacctgact caatatttt tccatatgc aacgatttc atccacgct gcgaggctc cgagaaggc aacgtttta atacattgt ctggctgga atgtctcgc agtgtgaca gtggcagtg gcgtacatc atgtctgta actaatctc aactggaag gaagcactt aaagtcgtt agagctggt cgcgcggta gccaacccc aacctgggc ttccaaaag caattacag gattttgaa gccacgcgg ttagcaaag gagcgccgc cgcctcaag gagcgctac ccaagttta gctttagta aacatagac agagaacag tgttctttg atgctcaat aactacgag gagttgctg caatctcgg tcaatttgc gaaggtcga tgcgcgttg ggcgaagaa tgtccaacg ggaatttgc agatcgaag aatcaaaac ccagctgct caccagccg cgttcacgg agacagagc agaagcaca agatcgcgt tcggagatg tgtcgaagt cctagtggc actagcgcc gctagcacc agcaccaga ccgcgctcg gggcccgca ggacttcgc agctataca ggtgcacta tacatatat gttgtaatg gttcagctc cgccatctc ggaccagtt cccgcgtag
- >TC003518 RC
atggggaat ggcatgaat aaggtgttg cctggcctt tatgtggga aattacagg gactcaaaa gacgctaac caactagcc aagcacaac ataacacac atccttgcc atccatgac tctgcgaaa agaatacat tccgataag cactatctt tgcgtgatg gcgtcggat tctccggat caaaacctg actcaatat ttttccata tgcaacgat ttcatccac gctgcgagg ctccgagaa ggcaacgtt ttaatacat tgtctggct ggaatgtct cgcagtgtg acagtggca gtggcgtac atcatgtct gtaactaat ctcaactgg aaggaagca cttaaagtc gttagagct ggtcgcgcg gtagccaac cccaacctg ggcttccaa aagcaatta caggatttt gaagccacg cggttagca aaggagcgc cgccgcctc aaggagcgc tacccaagt ttagcttta gtaaacata gacagagaa cagtgttct ttgatgctc aataactac gaggagttg ctgcaatct cggtcaatt tgcgaaggt cgatgcgcg ttgggcgaa gaatgtcca acgggaatt tgcagatcg aagaatcaa aacccagct gctcaccag ccgcgttca cggagacag agcagaagc acaagatcg cgttcggag atgtgtcga agtcctagt ggcactagc gccgctagc accagcacc agaccgcgc tcggggccc gcaggactt cgcagctat acaggttca gctccgcca tctcggacc agttcccgc gtagatatt tcttccatt ggtgtttcc ggaaacctt acagctcca acaacaccg caaacaagc cctccaatt tcaccgaaa cggcgagga acggtcgca gtgcctctg gctttgaaa gaagacgat tttgattta tag
- >TC003518 RA
cgggggcag acagagcgc cgccgccgt ttcagacga tgggatgcg agccgtttc cagttgagc cgtgtttag acatgtgat cgctgcccg agccagcaa ctgccgccc cggagacat ggggaatgg catgaataa ggtgttgcc tggccttta tgtgggaaa ttacaggga ctcaaaaga cgctaacca actagccaa gcacaacat aacacacat ccttgccat ccatgactc tgcgaaaag aatacattc cgataagca ctatctttg cgtgatggc gtcggattc tccggatca aaacctgac tcaatattt ttccatatg caacgattt catccacgc tgcgaggct ccgagaagg caacgtttt aatacattg tctggctgg aatgtctcg cagtgtgac agtggcagt ggcgtacat catgtctgt aactaatct caactggaa ggaagcact taaagtcgt tagagctgg tcgcgcggt agccaaccc caacctggg cttccaaaa gcaattaca ggattttga agccacgcg gttagcaaa ggagcgccg ccgcctcaa ggagcgcta cccaagttt agctttagt aaacataga cagagaaca gtgttcttt gatgctcaa taactacga ggagttgct gcaatctcg gtcaatttg cgaaggtcg atgcgcgtt gggcgaaga atgtccaac gggaatttg cagatcgaa gaatcaaaa cccagctgc tcaccagcc gcgttcacg gagacagag cagaagcac aagatcgcg ttcggagat gtgtcgaag tcctagtgg cactagcgc cgctagcac cagcaccag accgcgctc ggggcccgc aggacttcg cagctatac aggttcagc tccgccatc tcggaccag ttcccgcgt agatatttc ttccattgg tgtttccgg aaaccttac agctccaac aacaccgca aacaagccc tccaatttc accgaaacg gcgaggaac ggtcgcagt gcctctggc tttgaaaga agacgattt tgatttata gccctcgcc acaagaggt caaactctt caaaaattc taggggatg agtaggttt tggtgcgct tggttggag gttaaaagg gtgcaaatt tttgttggt tgtgatttt tgaaagaat ttatataca ccgtggcca ctagaagct tcgtgctta tatattacg aggaaacac tgcttatat tccatccgt gtcgtgttt agatccact ctagggttg tagaaaaat tatgtggta taatgtaac tcatcatat catcaaccc attttgatg actgatttt tttacgaat caaacacct aaacttaca atcattttt caattattc gtgatgctg gggttgaga attcgtcca cttacattt atcacaatc acaatatgt ttctttaaa attttgttt gattgaata cgtctacca tatgattaa gaccttccc atttaaact cgcaatttc tttagtatt tttattttc cattgcata gtttgtcac cgttgttac atgccaaaa agacattat catcgtcaa agatgtttt atatacaca catatattt ataaaataa tctaataaa aaagtaaaa caacacaag taaatcgga gctttttta aacaaacgt tagtacctg tactaaaaa cgactctca tatcagaaa tatgatgtt aaccacgaa ttgtcctcg atttttgtt gaattgggt tcgttaaac tcgaaaaaa atgtgtttc tcgttctga tttgtggca agcgcaaag ttgttttac tgaattaaa aaaaacctt tacatgttt ataatggtg aatatttta atgttaaca agagcaatg acttttccc cagtggaat tagagatag catgctcaa aaccaggct tagtgaaaa taattaagt agcacaaat gaatagtga taaaataca tattttatt agtgaagta ggttgttgt tcatggcgc tggccgccg gatttattg caataaagt acctacaca tttgttatt acggtttat tgtgcagtt taataatgg aacaattgc actgtagct atatttatg tgtatagaa ttgatgata aaaatatgc tacattatt agaaaacca ttcaaaaaa ttaaaaccg gtagcagta aagcaatgc tgagtgtac ctaattgca gttttgtta attggttga cccaaacat tgtttggtg ggtgttata atagtgata tttttttac aagtgacaa aaatgttga tactcgtac tttctacag attttacaa attgtttga agtgaaaat agcaccacg ctgatttct tgttagtta gttactgta aggcagaaa ggcgtaaga tcagtggag gtaatttta tggtctcta aaaacaaca tccttaaaa ttctttcac tgtattatg aagtgatag agggaaaca atcaagaat tgacgacgt aaatggacg ttaaattca agagtacaa ataatttta aaatgtttt tttgtattt atgaggctt tgtactaga ggcatagat gcacacact ttttgtttc tttatactt tattagtca cactttatt gtacaaaat actgtaaaa aatagccac gcaatatac gtagaagtg tccttattt gagttgcta tttttatat ttcaattat tgtacccaa gaatatatt gtcaattca cacctattg attaatcaa caatattga ctcagtggt ccacaaaaa gtgatataa tcataacct gcgttccag tccaatata cattatgtt ttaaataca tttttctag tgtgaaatg acaaaatgc ttggtcact atacataca aatatagaa acgttataa tttacgtat atttacgaa attaataaa taaagaaat tacag
- >TC003518 RB
cgggggcag acagagcgc cgccgccgt ttcagacga tgggatgcg agccgtttc cagttgagc cgtgtttag acatgtgat cgctgcccg agccagcaa ctgccgccc cggagacat ggggaatgg catgaataa ggtgttgcc tggccttta tgtgggaaa ttacaggga ctcaaaaga cgctaacca actagccaa gcacaacat aacacacat ccttgccat ccatgactc tgcgaaaag aatacattc cgataagca ctatctttg cgtgatggc gtcggattc tccggatca aaacctgac tcaatattt ttccatatg caacgattt catccacgc tgcgaggct ccgagaagg caacgtttt aatacattg tctggctgg aatgtctcg cagtgtgac agtggcagt ggcgtacat catgtctgt aactaatct caactggaa ggaagcact taaagtcgt tagagctgg tcgcgcggt agccaaccc caacctggg cttccaaaa gcaattaca ggattttga agccacgcg gttagcaaa ggagcgccg ccgcctcaa ggagcgcta cccaagttt agctttagt aaacataga cagagaaca gtgttcttt gatgctcaa taactacga ggagttgct gcaatctcg gtcaatttg cgaaggtcg atgcgcgtt gggcgaaga atgtccaac gggaatttg cagatcgaa gaatcaaaa cccagctgc tcaccagcc gcgttcacg gagacagag cagaagcac aagatcgcg ttcggagat gtgtcgaag tcctagtgg cactagcgc cgctagcac cagcaccag accgcgctc ggggcccgc aggacttcg cagctatac aggtgcact atacatata tgttgtaat ggttcagct ccgccatct cggaccagt tcccgcgta gatatttct tccattggt gtttccgga aaccttaca gctccaaca acaccgcaa acaagccct ccaatttca ccgaaacgg cgaggaacg gtcgcagtg cctctggct ttgaaagaa gacgatttt gatttatag ccctcgcca caagaggtc aaactcttc aaaaattct aggggatga gtaggtttt ggtgcgctt ggttggagg ttaaaaggg tgcaaattt ttgttggtt gtgattttt gaaagaatt tatatacac cgtggccac tagaagctt cgtgcttat atattacga ggaaacact gcttatatt ccatccgtg tcgtgttta gatccactc tagggttgt agaaaaatt atgtggtat aatgtaact catcatatc atcaaccca ttttgatga ctgattttt ttacgaatc aaacaccta aacttacaa tcatttttc aattattcg tgatgctgg ggttgagaa ttcgtccac ttacattta tcacaatca caatatgtt tctttaaaa ttttgtttg attgaatac gtctaccat atgattaag accttccca tttaaactc gcaatttct ttagtattt ttattttcc attgcatag tttgtcacc gttgttaca tgccaaaaa gacattatc atcgtcaaa gatgtttta tatacacac atatattta taaaataat ctaataaaa aagtaaaac aacacaagt aaatcggag cttttttaa acaaacgtt agtacctgt actaaaaac gactctcat atcagaaat atgatgtta accacgaat tgtcctcga tttttgttg aattgggtt cgttaaact cgaaaaaaa tgtgtttct cgttctgat ttgtggcaa gcgcaaagt tgttttact gaattaaaa aaaaccttt acatgttta taatggtga atattttaa tgttaacaa gagcaatga cttttcccc agtggaatt agagatagc atgctcaaa accaggctt agtgaaaat aattaagta gcacaaatg aatagtgat aaaatacat attttatta gtgaagtag gttgttgtt catggcgct ggccgccgg atttattgc aataaagta cctacacat ttgttatta cggtttatt gtgcagttt aataatgga acaattgca ctgtagcta tatttatgt gtatagaat tgatgataa aaatatgct acattatta gaaaaccat tcaaaaaat taaaaccgg tagcagtaa agcaatgct gagtgtacc taattgcag ttttgttaa ttggttgac ccaaacatt gtttggtgg gtgttataa tagtgatat ttttttaca agtgacaaa aatgttgat actcgtact ttctacaga ttttacaaa ttgtttgaa gtgaaaata gcaccacgc tgatttctt gttagttag ttactgtaa ggcagaaag gcgtaagat cagtggagg taattttat ggtctctaa aaacaacat ccttaaaat tctttcact gtattatga agtgataga gggaaacaa tcaagaatt gacgacgta aatggacgt taaattcaa gagtacaaa taattttaa aatgttttt ttgtattta tgaggcttt gtactagag gcatagatg cacacactt tttgtttct ttatacttt attagtcac actttattg tacaaaata ctgtaaaaa atagccacg caatatacg tagaagtgt ccttatttg agttgctat ttttatatt tcaattatt gtacccaag aatatattg tcaattcac acctattga ttaatcaac aatattgac tcagtggtc cacaaaaag tgatataat cataacctg cgttccagt ccaatatac attatgttt taaatacat ttttctagt gtgaaatga caaaatgct tggtcacta tacatacaa atatagaaa cgttataat ttacgtata tttacgaaa ttaataaat aaagaaatt acag
- >TC003518 RC
cgggggcag acagagcgc cgccgccgt ttcagacga tgggatgcg agccgtttc cagttgagc cgtgtttag acatgtgat cgctgcccg agccagcaa ctgccgccc cggagacat ggggaatgg catgaataa ggtgttgcc tggccttta tgtgggaaa ttacaggga ctcaaaaga cgctaacca actagccaa gcacaacat aacacacat ccttgccat ccatgactc tgcgaaaag aatacattc cgataagca ctatctttg cgtgatggc gtcggattc tccggatca aaacctgac tcaatattt ttccatatg caacgattt catccacgc tgcgaggct ccgagaagg caacgtttt aatacattg tctggctgg aatgtctcg cagtgtgac agtggcagt ggcgtacat catgtctgt aactaatct caactggaa ggaagcact taaagtcgt tagagctgg tcgcgcggt agccaaccc caacctggg cttccaaaa gcaattaca ggattttga agccacgcg gttagcaaa ggagcgccg ccgcctcaa ggagcgcta cccaagttt agctttagt aaacataga cagagaaca gtgttcttt gatgctcaa taactacga ggagttgct gcaatctcg gtcaatttg cgaaggtcg atgcgcgtt gggcgaaga atgtccaac gggaatttg cagatcgaa gaatcaaaa cccagctgc tcaccagcc gcgttcacg gagacagag cagaagcac aagatcgcg ttcggagat gtgtcgaag tcctagtgg cactagcgc cgctagcac cagcaccag accgcgctc ggggcccgc aggacttcg cagctatac aggttcagc tccgccatc tcggaccag ttcccgcgt agatatttc ttccattgg tgtttccgg aaaccttac agctccaac aacaccgca aacaagccc tccaatttc accgaaacg gcgaggaac ggtcgcagt gcctctggc tttgaaaga agacgattt tgatttata gccctcgcc acaagaggt caaactctt caaaaattc taggggatg agtaggttt tggtgcgct tggttggag gttaaaagg gtgcaaatt tttgttggt tgtgatttt tgaaagaat ttatataca ccgtggcca ctagaagct tcgtgctta tatattacg aggaaacac tgcttatat tccatccgt gtcgtgttt agatccact ctagggttg tagaaaaat tatgtggta taatgtaac tcatcatat catcaaccc attttgatg actgatttt tttacgaat caaacacct aaacttaca atcattttt caattattc gtgatgctg gggttgaga attcgtcca cttacattt atcacaatc acaatatgt ttctttaaa attttgttt gattgaata cgtctacca tatgattaa gaccttccc atttaaact cgcaatttc tttagtatt tttattttc cattgcata gtttgtcac cgttgttac atgccaaaa agacattat catcgtcaa agatgtttt atatacaca catatattt ataaaataa tctaataaa aaagtaaaa caacacaag taaatcgga gctttttta aacaaacgt tagtacctg tactaaaaa cgactctca tatcagaaa tatgatgtt aaccacgaa ttgtcctcg atttttgtt gaattgggt tcgttaaac tcgaaaaaa atgtgtttc tcgttctga tttgtggca agcgcaaag ttgttttac tgaattaaa aaaaacctt tacatgttt ataatggtg aatatttta atgttaaca agagcaatg acttttccc cagtggaat tagagatag catgctcaa aaccaggct tagtgaaaa taattaagt agcacaaat gaatagtga taaaataca tattttatt agtgaagta ggttgttgt tcatggcgc tggccgccg gatttattg caataaagt acctacaca tttgttatt acggtttat tgtgcagtt taataatgg aacaattgc actgtagct atatttatg tgtatagaa ttgatgata aaaatatgc tacattatt agaaaacca ttcaaaaaa ttaaaaccg gtagcagta aagcaatgc tgagtgtac ctaattgca gttttgtta attggttga cccaaacat tgtttggtg ggtgttata atagtgata tttttttac aagtgacaa aaatgttga tactcgtac tttctacag attttacaa attgtttga agtgaaaat agcaccacg ctgatttct tgttagtta gttactgta aggcagaaa ggcgtaaga tcagtggag gtaatttta tggtctcta aaaacaaca tccttaaaa ttctttcac tgtattatg aagtgatag agggaaaca atcaagaat tgacgacgt aaatggacg ttaaattca agagtacaa ataatttta aaatgtttt tttgtattt atgaggctt tgtactaga ggcatagat gcacacact ttttgtttc tttatactt tattagtca cactttatt gtacaaaat actgtaaaa aatagccac gcaatatac gtagaagtg tccttattt gagttgcta tttttatat ttcaattat tgtacccaa gaatatatt gtcaattca cacctattg attaatcaa caatattga ctcagtggt ccacaaaaa gtgatataa tcataacct gcgttccag tccaatata cattatgtt ttaaataca tttttctag tgtgaaatg acaaaatgc ttggtcact atacataca aatatagaa acgttataa tttacgtat atttacgaa attaataaa taaagaaat tacag
