| General information | |||
| TC033419
Former TC numbers If one gene is splitted or genes are merged, the original TC numbers are listed. One listed former TC number indicates the previous gene has be splitted, serveral former TC number represents a merged gene. |
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: 4894287..4905991 | |
| 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 | TC033419 | ||
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_02375 | 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 |
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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: 10.0% (includes death as pupa, adult)
eclosion not fulfilled (with pupal and adult features) - Number of animals showing the phenotype: 1 (3 dpi)gena triplet bristle pattern irregular - Penetrance (percentage of animals/eggs on slide): less than 30%
legs shape irregular - Penetrance (percentage of animals/eggs on slide): 30% - 50%
legs potentially swollen - Penetrance (percentage of animals/eggs on slide): 30% - 50%
affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): 30% - 50%
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)
- >TC033419 RA
MEASQESIS SRRDKLNTQ IIGDQKLTL INRECLLDA LQALYDECS LDALQKYDK QIAQFVSKY KNTLLEVKK LRVNETDFE IKDVIGRGH FGEVHVVKE KQTGDIYAM KTIRKCDSF EKTNFDVER DIMAFAASP WITSLQYAF QDSVYLYYV MEYHPGGDL LGLLYRQGG TLPESAATF YLAEIVLAL EDLHSMGFV HRDIKPDNI LLDRCGHLK LVDFGSAAK LNQQGAVDK GLPVGTPDY IAPEVLQSL ERNSKSHYG VSCDFWSLG ALAFELVVG KPPFSGQNT SVTYSRIIN HSNNLKFPQ DVCLTQAYV SLVRGLLTE AKARLRGPQ IKKHPLFKH IHFDSLRDQ VPPYVPKIS SMEDTSNFA DVGAKKTTP SMENFKKKT QFSGRNLPF IGFTFTPGA SSCGREFDR NSRAKDEII RNMREELEV VRRDLMKRE DFEEVKLVL EKKILEKNQ KLENIESVR EKLERDLAA SMAESTALK RTLEIERKD RAELERKAL DLIKSAKIK WETSERNKI ESLKLEIEQ QKEKIAQLG TTNKMLNEQ LQKAFKMEN KHKESIEAV EHLNRRSVI GLESRIEKV ALDSKQKIM SLQATLDET IKQKKELEF EIEKLKDSE DELRKKLSK SCDDYTVLN GAALEAEKL VKELGQKVG DFEREFEKA ESYKAKIKT LSREIDDFK AKIRDLENA NLLLKEENK SIEEFKNQI NSLKDTKVQ NERKINELE SKLQEEEQK CVTLKKELD NSQNLINES QQLKELRVK MWKMEKELG NAKIDKRIV ERELKDAEN EIKNLNEEI KNLKDKIEE NRKIHSAAL LELNNINEN LSLEMVKVR ESHKNLEEN LESEKNRNG EEKQLIVQL RDTISEKNK TISTIQSQL QSSRHELAT LEEKNQKLL ESNLKKSKQ FEEISNEKE KLSREVNET KRQLENAQT NLEALREAC LIIENQVIE YEKINSSIQ AEKSSLAEN TEKLIKDLC KAKEEIQEA KKATNEEKS LRLLAETKA KRLAEDVEL LQNECEAYK DQCFKFKEC SNNLSEELT LSEERVSDL EVTLKSYER QIEDFVSES NILKEEISQ YLTQMAALK ESNFKLKQQ IQDLKEARN VLVEKVEEL ERVLNGKVN YYEEREMKN ESIIKQQTK LIEYLQVKI DEHNSKKRT LTDKLFGSK KENVQPLVL PVNKDLEGQ LAKQKQINK NLQEEIYKL KAELNCRTP ITKVQRHKS EIVSPKTRS PQIISHKFI LKLCKNLTR CDHCAKQIM VGRDVNTCT KCHVNVHQN CVSSVKSTC GLPNTCVKN RKSESNVVI YEGWIKLLT KNNTWDKKY ACLTDSKLN VYLTKDKKS LTESFDLKL PNSRKKVVL EPLPSEIGV NVASTDLPF VMKVEVLVQ GGPMVSMVF MALSVEERN QWYKAFQEL FDEAAGQHL EAIFTLPDQ IYANCVLDL TKNIKVLGT DQGLFSYHN DQLVHIEGP TDIHHISII PNSNTVVMI SDLTSILIS CDLNHLINL TQCAPCTKP VLNYRTINI KEMDGFHYL QTSKFPKHQ LFCAATCRQ LVIVKYDCE NYEFVPLRI MDTAEPLGC ALFTEHSLI VGADKFFEI DLATFKAEE FLDGSDARL KFAVKCHKM QSFPVAILE ISQNPKEFL VCFNEFSVF VDEYGRSSR EEELKTSYL PIGFHFVAP FLYIIQYSG VEVLKVGGN VANVTVDLK NMRFLGESP NGIYLYHDG RVKLLEGRR VVDCDDVSS VSQSEAGDE DRFSFTSSM VQSLDGNSS DLENSSEEN LFLEHYEVI NRQTLDTAD VKTLTTVVE KECYKMKVT IDAFNDIFG WSFLLFTLY SSLLLLDYV SCVVRNEYK YKSDKMAEI ISSDLLQLC IFFVALIIL ILQCGRVRS EVVKILNVL KKIALRRQT TNGDLDELI VTLSTHYFP VFSACDYFT IETKTVLDV LGVTVTYLI VIIQFDDKI VQV
- >TC033419 RA
atggaagcg tcccaagag tccatcagt tcccgaagg gacaaactc aacactcaa ataatcggc gaccaaaaa ctcaccctc atcaaccga gaatgcctc ctggacgcc cttcaggcc ctttacgac gaatgcagt ctggacgct ttgcaaaaa tacgacaaa caaatcgcc caatttgtg agcaagtac aaaaacacg ctccttgag gtgaaaaaa ctgcgtgtg aatgagacc gatttcgag ataaaggat gtgatagga cgggggcac tttggggag gtgcatgtc gtgaaggag aagcaaact ggggatatt tacgccatg aagacgata cgaaagtgt gacagtttc gagaaaact aactttgat gttgagcga gatataatg gcctttgcg gcttcgccc tggatcaca agtctccag tacgcgttc caggacagt gtctacttg tactacgtg atggagtac caccctggg ggcgacttg ctggggctc ttgtaccgc caagggggc accttgccc gaaagcgcc gccaccttc tacctcgcc gaaattgtc ctcgctctg gaagacctg cactcgatg gggttcgtc caccgggac atcaaaccc gacaacatc ctcctcgac cgctgcggc cacctaaag ttggtcgac ttcggctca gccgccaaa ctcaaccag cagggggcc gtggacaag gggcttccg gtgggcacc cccgactac atcgccccc gaggtgctc cagtccctc gagcgcaac tccaagtcg cactacggc gtgtcgtgc gatttctgg tcgttgggg gccctcgcc ttcgagctg gtcgtgggc aagcccccg ttttcgggg caaaacacc tcagtcacc tactcgcgg ataatcaac cactcgaac aatttgaaa ttcccccag gatgtgtgc ttgacacaa gcctacgtc tccctggtt cggggtctc ctgactgag gcgaaggcc cgcctgagg gggccccag atcaagaaa caccccctt ttcaagcac attcatttc gattcgttg cgcgatcaa gtgcccccc tatgtcccc aaaatcagc tcgatggag gatacgagt aattttgca gacgtgggg gcgaagaaa accacccca agtatggag aacttcaag aagaagacg cagttttcc gggaggaat ttgcccttt attgggttt acgttcacc ccaggggcg agctcttgt gggagggag tttgatcgg aattcgagg gccaaggac gagattatt aggaatatg agggaagag ttggaggtg gtgaggagg gatttgatg aaacgggag gattttgag gaggtcaag ttggttttg gagaaaaag attctcgag aaaaaccag aagttggag aatattgaa agtgtgagg gagaagttg gagagggat ttggctgca agcatggcc gagagtacg gcattgaaa agaactctc gaaatcgag cgaaaagac cgcgccgaa ctcgaacgc aaagccctc gacttaata aaatcggca aaaatcaaa tgggaaaca tccgaacgc aacaaaatc gaatcatta aaactagaa attgagcaa caaaaagaa aaaatcgcc caacttgga accaccaac aaaatgcta aacgaacaa ttacaaaaa gctttcaaa atggaaaac aaacataaa gaatcgata gaagcagtc gaacactta aatcggcga agcgtaata ggacttgag tcacgaatt gaaaaagtc gccttagat tcgaaacaa aagattatg agtcttcaa gccacctta gacgaaaca attaaacaa aagaaggag ttagagttt gaaattgag aaattaaaa gacagtgag gacgaattg aggaaaaaa ctgtcgaaa tcttgtgat gattatact gttctgaat ggagcggct ctcgaggcc gaaaaactc gttaaagaa cttgggcag aaagtgggc gattttgag cgtgaattt gaaaaggcg gaaagctac aaagcgaag atcaagact ttaagtcgg gaaattgac gattttaag gcgaaaatt cgtgatttg gaaaatgcg aatttgtta ctcaaagag gaaaataag agtattgaa gagtttaag aatcagata aattcacta aaagacact aaggtacaa aatgagcga aaaatcaac gaactggag agcaagtta caagaagag gagcaaaaa tgtgtcaca ctgaagaaa gaattggac aattcgcaa aatctaatc aacgagagt caacagttg aaggaattg agggtaaaa atgtggaaa atggagaag gagttaggt aatgcaaag atcgacaaa cgaatagtt gagcgtgaa cttaaagat gcagaaaat gaaataaaa aatctcaat gaagaaata aaaaatttg aaagataag atagaagag aatcggaaa attcactcc gccgccttg ttagaattg aacaatatt aacgaaaac ctgtctttg gaaatggtc aaagtcagg gaatcccac aaaaacttg gaagaaaac ctcgaaagt gagaagaac cgaaacggt gaagagaaa caactaatc gtccaactc agagacact atcagcgaa aagaacaaa acaatttcc acaatccag tcccaactg caatcttcc agacacgaa ctcgcaacc cttgaagag aaaaaccag aaattgctc gaatcaaac ctgaaaaag tccaaacaa tttgaagaa atctcgaac gaaaaagag aaattatca agggaagtg aacgaaacg aagagacaa ttagaaaac gcccaaacg aatctagaa gccttgcgt gaggcttgc ctcatcatc gaaaatcaa gtgatcgag tacgaaaaa atcaactct tcgatccag gccgaaaaa tcgagtctt gccgaaaac accgagaaa ctgatcaag gatttgtgc aaagccaag gaggaaatc caggaggcg aagaaggcc accaatgag gagaaatcg ctgcggcta ttggccgaa actaaggcg aaacgattg gcggaggat gtggagttg ttgcagaat gagtgtgag gcttacaaa gatcagtgt tttaaattt aaagagtgt tcgaataat ttgagtgaa gagctgacg ctttcggag gagagggtc tccgatttg gaggttacc ctcaaatcg tacgagagg caaattgag gatttcgtc tccgaaagt aacatcttg aaagaggag atttcgcaa tatttgact caaatggcc gccctcaaa gagtcgaat ttcaaactc aaacagcaa attcaggac ttgaaggag gcgcggaat gtactagtg gagaaggtt gaggagttg gagagggtc ctcaatggt aaagtcaac tattacgaa gaacgggag atgaagaac gagtcgatt attaagcaa caaaccaag ctgattgag tatctccaa gttaagatt gacgaacac aacagtaaa aaacgaaca ctgactgat aaattgttc ggttcgaag aaagaaaat gtgcaacct ttggtcctt ccggtcaat aaagacctg gaaggacaa ttggcgaaa caaaaacaa atcaacaaa aacctacaa gaggaaata tacaaattg aaggcggag cttaactgt cgcacacca atcacaaaa gtgcaacgt cacaaatcg gaaatcgtg tcacctaag accagatcg cctcaaata atatcacac aaatttata ctaaagctt tgtaaaaac ttgacacgt tgtgaccac tgcgccaaa caaatcatg gtgggccgt gacgtcaac acatgtacg aaatgtcac gtcaacgtg catcaaaat tgtgtcagt tctgtcaaa agtacttgc ggcttgcct aacacatgt gtcaaaaac cgtaaaagc gaatcaaat gtcgtaatt tacgaaggt tggattaaa ttacttact aaaaacaac acttgggac aaaaaatac gcttgtttg actgattcc aaattgaat gtttattta actaaagat aagaaaagt ctgacggaa tcgtttgat ttgaaattg ccaaattcg aggaagaaa gttgttttg gaaccgctg ccgtctgaa atcggggtg aatgttgcc agtacggac ctgccgttt gtgatgaaa gtggaggtt ttggtacag ggtgggccg atggtttcg atggttttt atggccttg tcggtggag gagaggaat cagtggtat aaagcattt caagagctc ttcgacgaa gccgccggc caacacctg gaggccatc ttcaccctc cctgaccaa atttacgcc aactgcgtc ctggatctc accaaaaac atcaaagtt ttaggcacc gaccaaggt ctattttcc taccacaac gatcaactt gtacatatc gagggcccc accgacata catcacata tcaataatc cccaactct aacaccgtg gtcatgatc tcagaccta actagcatt ttaatctca tgtgactta aaccacctc attaattta acccaatgc gcaccttgt actaaacca gtccttaat tatcgcaca atcaatatc aaagaaatg gacggtttt cactacttg caaacgtcg aaattcccc aaacaccaa ttattttgt gccgcgact tgtcgccag ttggtaatc gtgaaatac gactgcgaa aattacgaa ttcgttccg ttgcggatt atggacacg gccgaaccg ctggggtgt gcgttattt acggaacac tcgctgatt gtgggcgct gataaattt ttcgaaatc gatttagcc actttcaag gccgaggaa ttcctcgat gggtcagac gcgaggttg aaattcgca gtcaagtgt cacaaaatg cagtcgttt ccggtcgca attttggaa atttcgcaa aatccgaag gagtttttg gtctgtttt aacgaattt tcggtgttt gtggacgag tatgggcgg agttcgcgc gaggaggag ttgaaaacg agttatctg ccaattggg ttccacttt gtggcgcct tttttgtat attattcag tacagtgga gtggaagtt ttgaaagtt ggtggaaac gtggcgaat gtgacggtt gatttgaaa aacatgcgg tttttgggg gaaagcccg aacgggatt tatctctat catgacgga agagttaag ttgttggag gggcgcaga gtggttgat tgtgacgat gtcagttcg gtcagccaa tcggaggcc ggggatgag gataggttt agttttacg agctcgatg gtgcaaagt ctcgatggg aattccagc gatttggag aacagtagc gaggagaat ttgtttttg gaacattac gaagtcata aatagacaa acgcttgac actgcagat gtcaaaact ttgacaaca gtcgtcgag aaagagtgt tacaaaatg aaagtcacc attgatgcg tttaatgat attttcggc tggtctttc ctacttttt acgctatat tccagcctt ttacttctc gactacgtc agttgtgtg gtaaggaat gaatataag tacaaatcg gacaaaatg gccgaaatc atatcatca gatttgcta caactttgc atttttttc gtcgccctg attattttg atattgcaa tgcgggaga gtacgatca gaagtcgta aaaatctta aatgtgctt aagaaaatt gccttaagg cgccaaaca accaatggc gacttggac gaacttatc gtcacgctt tcaactcat tattttccg gttttcagc gcttgtgat tatttcact attgaaact aagaccgtg ttggatgtt ttgggggtc actgtgacc tacctgatc gttattata caatttgac gataaaata gtgcaagtt tag
- >TC033419 RA
tgtaacggt tttttcaaa tttcaatca aacccgtgt ttactttcc acttttccg tgtttaaaa tgtgtgtat gagtgtgtt tttgtcatg gaagcgtcc caagagtcc atcagttcc cgaagggac aaactcaac actcaaata atcggcgac caaaaactc accctcatc aaccgagaa tgcctcctg gacgccctt caggccctt tacgacgaa tgcagtctg gacgctttg caaaaatac gacaaacaa atcgcccaa tttgtgagc aagtacaaa aacacgctc cttgaggtg aaaaaactg cgtgtgaat gagaccgat ttcgagata aaggatgtg ataggacgg gggcacttt ggggaggtg catgtcgtg aaggagaag caaactggg gatatttac gccatgaag acgatacga aagtgtgac agtttcgag aaaactaac tttgatgtt gagcgagat ataatggcc tttgcggct tcgccctgg atcacaagt ctccagtac gcgttccag gacagtgtc tacttgtac tacgtgatg gagtaccac cctgggggc gacttgctg gggctcttg taccgccaa gggggcacc ttgcccgaa agcgccgcc accttctac ctcgccgaa attgtcctc gctctggaa gacctgcac tcgatgggg ttcgtccac cgggacatc aaacccgac aacatcctc ctcgaccgc tgcggccac ctaaagttg gtcgacttc ggctcagcc gccaaactc aaccagcag ggggccgtg gacaagggg cttccggtg ggcaccccc gactacatc gcccccgag gtgctccag tccctcgag cgcaactcc aagtcgcac tacggcgtg tcgtgcgat ttctggtcg ttgggggcc ctcgccttc gagctggtc gtgggcaag cccccgttt tcggggcaa aacacctca gtcacctac tcgcggata atcaaccac tcgaacaat ttgaaattc ccccaggat gtgtgcttg acacaagcc tacgtctcc ctggttcgg ggtctcctg actgaggcg aaggcccgc ctgaggggg ccccagatc aagaaacac ccccttttc aagcacatt catttcgat tcgttgcgc gatcaagtg cccccctat gtccccaaa atcagctcg atggaggat acgagtaat tttgcagac gtgggggcg aagaaaacc accccaagt atggagaac ttcaagaag aagacgcag ttttccggg aggaatttg ccctttatt gggtttacg ttcacccca ggggcgagc tcttgtggg agggagttt gatcggaat tcgagggcc aaggacgag attattagg aatatgagg gaagagttg gaggtggtg aggagggat ttgatgaaa cgggaggat tttgaggag gtcaagttg gttttggag aaaaagatt ctcgagaaa aaccagaag ttggagaat attgaaagt gtgagggag aagttggag agggatttg gctgcaagc atggccgag agtacggca ttgaaaaga actctcgaa atcgagcga aaagaccgc gccgaactc gaacgcaaa gccctcgac ttaataaaa tcggcaaaa atcaaatgg gaaacatcc gaacgcaac aaaatcgaa tcattaaaa ctagaaatt gagcaacaa aaagaaaaa atcgcccaa cttggaacc accaacaaa atgctaaac gaacaatta caaaaagct ttcaaaatg gaaaacaaa cataaagaa tcgatagaa gcagtcgaa cacttaaat cggcgaagc gtaatagga cttgagtca cgaattgaa aaagtcgcc ttagattcg aaacaaaag attatgagt cttcaagcc accttagac gaaacaatt aaacaaaag aaggagtta gagtttgaa attgagaaa ttaaaagac agtgaggac gaattgagg aaaaaactg tcgaaatct tgtgatgat tatactgtt ctgaatgga gcggctctc gaggccgaa aaactcgtt aaagaactt gggcagaaa gtgggcgat tttgagcgt gaatttgaa aaggcggaa agctacaaa gcgaagatc aagacttta agtcgggaa attgacgat tttaaggcg aaaattcgt gatttggaa aatgcgaat ttgttactc aaagaggaa aataagagt attgaagag tttaagaat cagataaat tcactaaaa gacactaag gtacaaaat gagcgaaaa atcaacgaa ctggagagc aagttacaa gaagaggag caaaaatgt gtcacactg aagaaagaa ttggacaat tcgcaaaat ctaatcaac gagagtcaa cagttgaag gaattgagg gtaaaaatg tggaaaatg gagaaggag ttaggtaat gcaaagatc gacaaacga atagttgag cgtgaactt aaagatgca gaaaatgaa ataaaaaat ctcaatgaa gaaataaaa aatttgaaa gataagata gaagagaat cggaaaatt cactccgcc gccttgtta gaattgaac aatattaac gaaaacctg tctttggaa atggtcaaa gtcagggaa tcccacaaa aacttggaa gaaaacctc gaaagtgag aagaaccga aacggtgaa gagaaacaa ctaatcgtc caactcaga gacactatc agcgaaaag aacaaaaca atttccaca atccagtcc caactgcaa tcttccaga cacgaactc gcaaccctt gaagagaaa aaccagaaa ttgctcgaa tcaaacctg aaaaagtcc aaacaattt gaagaaatc tcgaacgaa aaagagaaa ttatcaagg gaagtgaac gaaacgaag agacaatta gaaaacgcc caaacgaat ctagaagcc ttgcgtgag gcttgcctc atcatcgaa aatcaagtg atcgagtac gaaaaaatc aactcttcg atccaggcc gaaaaatcg agtcttgcc gaaaacacc gagaaactg atcaaggat ttgtgcaaa gccaaggag gaaatccag gaggcgaag aaggccacc aatgaggag aaatcgctg cggctattg gccgaaact aaggcgaaa cgattggcg gaggatgtg gagttgttg cagaatgag tgtgaggct tacaaagat cagtgtttt aaatttaaa gagtgttcg aataatttg agtgaagag ctgacgctt tcggaggag agggtctcc gatttggag gttaccctc aaatcgtac gagaggcaa attgaggat ttcgtctcc gaaagtaac atcttgaaa gaggagatt tcgcaatat ttgactcaa atggccgcc ctcaaagag tcgaatttc aaactcaaa cagcaaatt caggacttg aaggaggcg cggaatgta ctagtggag aaggttgag gagttggag agggtcctc aatggtaaa gtcaactat tacgaagaa cgggagatg aagaacgag tcgattatt aagcaacaa accaagctg attgagtat ctccaagtt aagattgac gaacacaac agtaaaaaa cgaacactg actgataaa ttgttcggt tcgaagaaa gaaaatgtg caacctttg gtccttccg gtcaataaa gacctggaa ggacaattg gcgaaacaa aaacaaatc aacaaaaac ctacaagag gaaatatac aaattgaag gcggagctt aactgtcgc acaccaatc acaaaagtg caacgtcac aaatcggaa atcgtgtca cctaagacc agatcgcct caaataata tcacacaaa tttatacta aagctttgt aaaaacttg acacgttgt gaccactgc gccaaacaa atcatggtg ggccgtgac gtcaacaca tgtacgaaa tgtcacgtc aacgtgcat caaaattgt gtcagttct gtcaaaagt acttgcggc ttgcctaac acatgtgtc aaaaaccgt aaaagcgaa tcaaatgtc gtaatttac gaaggttgg attaaatta cttactaaa aacaacact tgggacaaa aaatacgct tgtttgact gattccaaa ttgaatgtt tatttaact aaagataag aaaagtctg acggaatcg tttgatttg aaattgcca aattcgagg aagaaagtt gttttggaa ccgctgccg tctgaaatc ggggtgaat gttgccagt acggacctg ccgtttgtg atgaaagtg gaggttttg gtacagggt gggccgatg gtttcgatg gtttttatg gccttgtcg gtggaggag aggaatcag tggtataaa gcatttcaa gagctcttc gacgaagcc gccggccaa cacctggag gccatcttc accctccct gaccaaatt tacgccaac tgcgtcctg gatctcacc aaaaacatc aaagtttta ggcaccgac caaggtcta ttttcctac cacaacgat caacttgta catatcgag ggccccacc gacatacat cacatatca ataatcccc aactctaac accgtggtc atgatctca gacctaact agcatttta atctcatgt gacttaaac cacctcatt aatttaacc caatgcgca ccttgtact aaaccagtc cttaattat cgcacaatc aatatcaaa gaaatggac ggttttcac tacttgcaa acgtcgaaa ttccccaaa caccaatta ttttgtgcc gcgacttgt cgccagttg gtaatcgtg aaatacgac tgcgaaaat tacgaattc gttccgttg cggattatg gacacggcc gaaccgctg gggtgtgcg ttatttacg gaacactcg ctgattgtg ggcgctgat aaatttttc gaaatcgat ttagccact ttcaaggcc gaggaattc ctcgatggg tcagacgcg aggttgaaa ttcgcagtc aagtgtcac aaaatgcag tcgtttccg gtcgcaatt ttggaaatt tcgcaaaat ccgaaggag tttttggtc tgttttaac gaattttcg gtgtttgtg gacgagtat gggcggagt tcgcgcgag gaggagttg aaaacgagt tatctgcca attgggttc cactttgtg gcgcctttt ttgtatatt attcagtac agtggagtg gaagttttg aaagttggt ggaaacgtg gcgaatgtg acggttgat ttgaaaaac atgcggttt ttgggggaa agcccgaac gggatttat ctctatcat gacggaaga gttaagttg ttggagggg cgcagagtg gttgattgt gacgatgtc agttcggtc agccaatcg gaggccggg gatgaggat aggtttagt tttacgagc tcgatggtg caaagtctc gatgggaat tccagcgat ttggagaac agtagcgag gagaatttg tttttggaa cattacgaa gtcataaat agacaaacg cttgacact gcagatgtc aaaactttg acaacagtc gtcgagaaa gagtgttac aaaatgaaa gtcaccatt gatgcgttt aatgatatt ttcggctgg tctttccta ctttttacg ctatattcc agcctttta cttctcgac tacgtcagt tgtgtggta aggaatgaa tataagtac aaatcggac aaaatggcc gaaatcata tcatcagat ttgctacaa ctttgcatt tttttcgtc gccctgatt attttgata ttgcaatgc gggagagta cgatcagaa gtcgtaaaa atcttaaat gtgcttaag aaaattgcc ttaaggcgc caaacaacc aatggcgac ttggacgaa cttatcgtc acgctttca actcattat tttccggtt ttcagcgct tgtgattat ttcactatt gaaactaag accgtgttg gatgttttg ggggtcact gtgacctac ctgatcgtt attatacaa tttgacgat aaaatagtg caagtttag gcaaaaaat attcttact ggtcctacg ctcaaataa ttttattcg ctcgcttgt tgatttgtt ttaactgtt taaaacttg ccacaattg tcaagaata aataaatga cgttact
