| General information | |||
| TC034585 | 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: 1429027..1437300 | |
| 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 | TC034585 | ||
| General information | ||||
| iB_00474 | 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: 0.0% (includes death as pupa, adult)
potentially confused/crosscontaminated with : adults of..., iB # (comma separated)Adult morphology
potentially confused/crosscontaminated with : adults of..., iB # (comma separated)
Analysis of embryonic musculature and early development
number of eggs/embryos/larvae on slide: > 50
embryo/egg not developed/not fertilized -
Penetrance (percentage of animals/eggs on slide): more than 80%
affected eggs/embryos/larvae total percentage of affected eggs/embryos/larvae on slide - Penetrance (percentage of animals/eggs on slide): more than 80%
potentially confused/crosscontaminated with : adults of..., iB # (comma separated) (egglay)
affected eggs/embryos/larvae total percentage of affected eggs/embryos/larvae on slide - Penetrance (percentage of animals/eggs on slide): more than 80%
potentially confused/crosscontaminated with : adults of..., iB # (comma separated) (egglay)
Analysis of larval stage 1 cuticle
number of eggs/embryos/larvae on slide: > 50
egg without cuticle embryo/egg without visible larval cuticle in cuticle preparation - Penetrance (percentage of animals/eggs on slide): more than 80%
egg without cuticle embryo/egg without visible larval cuticle in cuticle preparation - Penetrance (percentage of animals/eggs on slide): more than 80%
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: 0.0% (includes death as larva, prepupa, pupa)
Lethalities 22 days after larval injection: 0.0% (includes death as larva, prepupa, pupa, adult)
Pupal morphology
remarks: later than schedule, hours 24
Adult morphology
remarks: later than schedule, hours 24
Fertility
remarks: later than schedule, hours 24
- >TC034585 RA
MYGSFNELY SGREIWNSN NWRNRNSRS GVRRNRSPS TSSERSFPG RNRSGFVPG QQDQRGGRR PNQNARKMG FKALQDLLN KDSEDLILE LSNIKRGIQ DYIKTSLSD DGIVLFVKV LKKVSDSAF VENKYKVLQ LYLDEVFVD KLVKYVALL PMQSDLDRT RNQFFWENT DEFWNNVIG ICQDIYNAI PTFGFKVLP NILKSLLVC FPAFQDQHK QHISDGIRD SVNKLNESL QIMIAEEEE KKKASIQKK VSTLIEEEP PDDFHQISV YPNSLELLG GTRVFLRKN IVEGPYKDV HHYLDVQFR LLREDFVGP LRNGIATYR TESTQKQRI ENVNIYKKV QFLNQETIN EQYCIRLRF DFSRKTKKF RFEDSKKFM FGSLLCFTC DNFKSLLFG KIAQRDVKD LEKGELIVG FDQNVLVDY KANYLMIEC GIYFEPYFH VLNVLKNIN TDEFPMEKY IIRVDPTPQ PPEYLNDPT PRKISIRGH DFFPLLNWP NTNFYTFNQ SQLRAFRAA LTQEFSVIQ GPPGTGKTF LGLKIAHTL LQNQAIWFK KTPMLVICY TNHALDQFL EGLALATDR IIRIGGQSR NQNVAKFNL RNIKFRTNP AVLQQRHGV KVLLLQIQN ITENLKEID FYHSIRDFK VFIDVIPNF ITTWFHRAT IDELLDWLL PEYENEDLD STNEVKSPR SVAEVQDKV ANLQIEDEM FDENLSDYE IEEDENMKI ELDDLFDEV NAMAVQCSI PLITLSSLQ QRISALERT FQQLKQKEE ITMEDIFEE DRIVFELYK LKFRYDSLN FRLTEGKQM RFMKRRVPD WSEFTNPHK LSPDDRWNL YFFCLNLYR QKLNEKLSV LNEQFREQH KIYEEMRDI ENTKAMKNV LVVGMTTTG AARLRSSLQ TLKSPIVIV EEAAEILEA HIVSSLTKH CKHLILIGD HQQLKPSTA SYNIEKFYN LGISLFERM VVNRIQLNT LNVQHRMRP EIASLVSPT IYPTLQDHP SVNDRPDIK GVDNCLFFI DHKHPEANC EGKSKKNYH EVDFLIYFA RHLILNGYE PGNITILAA YLGQMFELQ KERRKHNEL LANVRIAVL DNYQGEECD IILLSLVRN NEENKIGFL SIENRVCVA LSRARNGFY LMGNMDQLC AASQLWREI YKTFERQNA IGPHLALRC QVHPDKVTY VASGKDFLN ISEGGCNQK CGADLNCGH QCTSLCHVL NRDHASYRC LETCGKKLC DKNELHLCQ RRCYQECGP CTYKVSRTL KCGHVVNLE CHLDPATYS CEELVPTQL PCGHNADKP CHCNPETFP CPFPCEKQV EPCGHACTR NCHVRQDPD HLEYKCKKR CAKNQKNCK AEPEKHKCK KLCFEDCDA CMVRVRKQR TICPHFYDV PCSSNVDDI SCEKPCTKI LPCNHKCKN KCNQPCGNC KEKVEKQIP VCAHTIKVK CSEEPLRKF CTKKCPLTL PCGHPCTKK CNESCTPTC TVLATCALK APCGHTVTK LPCHLAHGA LDPKTLLQH CNHPCGGQL LCEHTCSGT CGECSQGRI HKRCDAKCG NPLVCNHIC EFPCRQACR PCVRPCPYK CKHSACGKR CGDICVPCK ESCPRRCKH RRCSKYCGE ICDVPPCQE PCSKLLACG HQCIGFCGD PCPQKCRIC DEDEVTEVF FGFEAEDDA RFVLLEDCG HIFENTGLE TWLGQDDGE VIKFKECPR CKTAIKTTA RFSDYVKKA KADVAKVKK NLHGTENEL NMKRQELAR LLLKVRTNA QDVKCSNFF KDILGAMET RLRISLMGK RQRINKFEL NAIESKLQI IEHIVDIGI NCPTALSKN RCSSQVKFI LKILEREDN YVTDQEIDD IPYELNRLS RIAELEEIE SSPNFLQRY SNSSTVRRV TEQIEGFLF GFGKFYDQD EQTVKELLQ ELSRLVASG IGISEQEKR EIVKAMGFR QGHWFKCPN GHPYCIADC GGAMVESKC PECACKIGG GSHTLRADN ALASEMDGA QFPAWSETA NNMGNFLLD FDD
- >TC034585 RA
atgtacgga agttttaac gaattatat tcaggaaga gaaatttgg aattccaac aactggaga aaccgtaat tctaggagc ggagtacga cgaaaccga agtccaagt accagttcc gagcggtcc tttcctggt aggaatcgt tctggattc gtcccagga caacaagac caaagaggt ggcagaaga cctaaccaa aatgcaaga aaaatgggt ttcaaagcc ctgcaagat ttgttaaac aaggattcc gaggattta atcctagaa ttgtccaac attaaaaga ggcatacag gactacatc aaaacgagt ttgtccgac gatggcatt gtgctgttc gtcaaagtt ttgaagaaa gtgtcggac tcggccttt gttgaaaac aagtacaag gttttacaa ttatatctg gatgaagta tttgttgac aaattggta aagtatgtt gctttattg ccaatgcaa agcgattta gaccgaaca aggaatcaa tttttttgg gaaaacacc gatgagttt tggaacaat gttatagga atttgccag gatatctat aatgcaatc cccacattt gggttcaaa gttttacca aatatactg aaaagtttg ctcgtctgt ttccccgcg tttcaagac caacacaag caacatatt tcggatggg ataagggac agtgtgaac aaactcaat gaaagtttg caaatcatg attgcagag gaagaagaa aagaagaaa gcttctata caaaagaaa gtttcaaca ttaattgag gaagaacca ccggatgat tttcatcaa atttctgtc taccccaat tcacttgaa ctccttgga ggcactcga gtttttctg cgtaaaaac atagtagaa ggaccttat aaagacgta caccactac ttggacgtc cagtttaga ctgctaaga gaagacttt gttgggcct ttgagaaat gggatagcc acgtaccga acagaatca acccagaag cagcgaatt gaaaatgtc aatatttac aaaaaagta cagttttta aatcaagag acaatcaac gaacaatac tgcattcga ttaaggttc gacttttca cgcaaaaca aagaaattt cgctttgaa gactccaag aaattcatg tttggttct ctcctctgc ttcacctgc gacaacttc aagtcacta ctctttggc aaaattgct cagcgagac gtgaaagac cttgaaaag ggggaactg attgtaggt tttgatcaa aatgtgctc gtggactac aaagcgaac tatttgatg atcgaatgc ggaatctac ttcgagccg tatttccac gtcctcaac gtcctcaaa aacataaac actgacgaa ttcccgatg gaaaaatac atcattcgc gtggacccc acacctcaa ccgccggaa tacctcaac gacccaacc ccccgtaaa atctcaatc cgtggtcat gattttttc cctctcctc aactggccc aacaccaac ttctacact ttcaaccaa tcgcaacta agggccttc cgggcagct ttaacgcaa gagttctcc gttattcaa ggcccacct ggcactggg aagaccttc cttggcctc aaaatcgcc cacactttg ctgcaaaat caggccatt tggttcaag aaaaccccg atgttggtc atctgctac accaaccat gccctcgac cagttcttg gagggtttg gctctggcc actgacaga atcattcga attggggga cagagcaga aaccaaaat gtggcaaag ttcaatttg agaaatata aagttccgg acaaacccg gccgtttta caacaaaga cacggagtg aaagtcctc ttgctacaa atccagaat ataacggaa aacttgaaa gaaatcgat ttttatcac agcatcaga gatttcaaa gtttttatt gatgtgata ccgaatttt attacaaca tggtttcat cgcgctaca atagacgaa ctactggac tggttgctg cccgaatat gaaaatgaa gacttggat tcgactaat gaagttaaa agtcctagg agtgttgca gaggtccag gacaaagtc gctaacttg cagattgaa gatgaaatg tttgatgaa aatttgtcg gattatgaa atcgaagaa gatgaaaat atgaaaatt gagttggat gatcttttc gatgaagtg aatgcaatg gcggttcag tgcagcatt ccccttatt acgctctcc tcacttcag caaaggatt tctgcacta gaaagaact ttccagcaa ctgaagcaa aaggaagag ataactatg gaggatatt ttcgaagaa gatcgaatt gtcttcgaa ttgtataaa ctgaaattt cgttacgac agtttaaat tttagacta accgagggc aagcaaatg cgtttcatg aaaagaaga gttcctgat tggtcagag ttcacgaat cctcacaaa ttgtcaccc gacgatcgc tggaattta tactttttt tgtttaaat ttatacaga cagaaatta aatgagaag ctgagcgta ctaaatgag cagttccgc gaacaacac aaaatctat gaagaaatg cgggacatt gaaaacacg aaagcaatg aaaaacgtg cttgttgtt ggtatgacg accactgga gcagcccgt ctaagatct tcgttgcaa acgttgaaa agtccaatc gttattgtc gaggaagca gcggagatt ctagaagct cacatcgtt tcctctttg accaaacat tgcaaacac ttgattctc atcggagac catcagcag ttgaaaccg agcactgcg agttataac atagaaaag ttttacaat ttggggata agtctattt gagagaatg gttgtgaac agaatccag ttgaatacc ctaaacgtg caacacaga atgaggccg gagattgcc agtttggtc tcgccgacc atttatcca accctgcaa gatcacccg tctgtgaac gacagacct gatataaaa ggggtcgat aattgcttg tttttcatc gaccacaag caccctgaa gcaaattgt gaaggcaaa agcaagaag aattaccat gaagttgac ttcttgatc tatttcgca aggcatctt attctgaac ggttacgaa cccggaaac atcacaatc ctggcggct tacttgggg caaatgttc gagttgcag aaagagagg cgcaaacac aatgagctt ctagcgaac gtaagaatt gcagttttg gataactat caaggtgaa gaatgcgat attatccta ctttcgctg gttcgaaac aacgaggaa aacaaaatc gggttccta tcaattgaa aacagagtt tgtgtggcg ctttcaaga gccagaaac ggcttctat ctcatgggg aacatggac cagttgtgt gccgctagt caactttgg cgtgagatt tacaagact tttgaacgc cagaatgcc attggaccc cacctagcc ctacggtgc caagtgcac ccagataag gtcacgtat gtggcaagc ggaaaggat tttctgaat atatcagaa ggagggtgc aaccagaaa tgtggggct gatttgaat tgcggtcat cagtgcact tctctttgc cacgtgtta aatcgagat catgcaagc tacaggtgc cttgaaacg tgcgggaaa aaattgtgc gataaaaat gaactacat ttatgccaa agaaggtgt taccaagag tgtggtcct tgcacttac aaggtgtca aggacccta aaatgtggg catgttgtg aacttggag tgtcacctg gatcctgca acttacagt tgtgaggag ctcgtgcct acgcaactt ccctgtggt cacaacgct gacaagcct tgccactgc aatccggaa acttttcct tgcccgttc ccctgcgaa aaacaggtg gaaccttgt ggccatgcg tgcactaga aattgccac gttcggcag gatccagat cacttagaa tacaaatgt aagaaacgt tgcgctaaa aaccaaaag aattgcaaa gccgaaccc gaaaaacac aagtgcaag aaactatgt tttgaggat tgtgacgcc tgtatggtc cgtgttagg aaacagcga accatctgt ccacatttt tacgacgtg ccttgttcc agtaacgtg gacgatata agctgcgaa aagccttgc accaaaata ttgccttgc aatcacaaa tgcaaaaat aaatgcaat caaccgtgc ggaaactgc aaagaaaag gttgagaaa caaatcccg gtatgtgcc catacaatc aaagtaaaa tgtagcgaa gaacctctc cgaaaattc tgtaccaaa aaatgtcct ctgacacta ccctgtggc cacccttgc acgaaaaaa tgcaacgag tcttgcacc ccaacttgc accgttctg gccacctgt gcattgaaa gccccttgt ggccacacc gtgaccaag ttaccgtgc cacctagcc catggtgcc ctcgaccct aaaaccctg ctacaacac tgcaaccac ccttgcggt ggccagtta ctgtgcgag cacacctgc tcgggaacc tgcggggag tgctcgcag ggccgcatc cacaaacgc tgcgacgca aaatgcggg aaccccctg gtgtgcaac cacatctgc gagttcccc tgcagacag gcctgcagg ccctgcgtg cgcccttgc ccctacaag tgcaagcac agcgcctgt gggaagcgc tgcggcgat atctgcgtc ccttgcaag gagagctgc ccccgcagg tgcaagcac cgaaggtgc tccaagtac tgcggggaa atctgcgac gtgccccct tgtcaagag ccttgcagc aaattactg gcatgtggc caccaatgt atcgggttc tgcggcgat ccttgccct caaaaatgc cgcatttgt gacgaagac gaggtgacc gaagtgttt ttcgggttc gaggctgaa gacgatgcc cggtttgtg ctcctggag gactgcggc cacattttc gaaaatacg ggactggaa acgtggctg gggcaagac gacggggaa gtgatcaag ttcaaggag tgtccgcgg tgcaaaacg gcgatcaaa acaacggcc cgattcagt gattatgtg aagaaagcg aaggccgat gtggccaaa gttaagaaa aatctgcac gggactgag aatgagttg aacatgaag aggcaagaa ctggcgaga ctgctacta aaagttagg acgaatgca caagatgtg aaatgttcc aacttcttt aaagatatt ttgggcgcg atggaaacc cgtctgcga atttctcta atgggcaag cgccaacgt atcaacaaa tttgagttg aatgcaatt gaatccaaa ctccaaatc atcgaacac atcgttgac attggcata aactgcccc acggctctt tcaaaaaat cgctgctcc tctcaggtc aaattcata ctcaagatt ttggaacgt gaggacaac tacgtaacg gatcaagaa atagacgat atcccgtac gaattgaac cgtttgtca cgaatcgcc gaacttgaa gaaatcgaa agttcgcca aacttcctc caacgatat tcaaattct tccacagtt aggcgcgtt accgaacaa attgaaggc tttctcttc ggtttcggc aagttttat gaccaggat gaacaaacg gtcaaggaa ttattgcaa gagttgagt cgattggtc gcgtcggga ataggaatt tcagaacag gaaaaaagg gaaattgtt aaagccatg ggtttcagg caagggcat tggttcaag tgccccaat ggccaccct tactgcata gctgactgt gggggcgca atggtggag tcaaaatgc ccggaatgt gcgtgcaaa ataggggga ggtagtcac acactgcgc gccgataat gcgttggct tctgaaatg gatggagcc cagtttccc gcttggtcg gaaactgca aacaatatg ggcaatttt ttgttagat ttcgatgat tga
- >TC034585 RA
cgcggacgc ggtggtgga aaccctaac tggcggtcc cgtcgagtt gataatgaa atgtacgga agttttaac gaattatat tcaggaaga gaaatttgg aattccaac aactggaga aaccgtaat tctaggagc ggagtacga cgaaaccga agtccaagt accagttcc gagcggtcc tttcctggt aggaatcgt tctggattc gtcccagga caacaagac caaagaggt ggcagaaga cctaaccaa aatgcaaga aaaatgggt ttcaaagcc ctgcaagat ttgttaaac aaggattcc gaggattta atcctagaa ttgtccaac attaaaaga ggcatacag gactacatc aaaacgagt ttgtccgac gatggcatt gtgctgttc gtcaaagtt ttgaagaaa gtgtcggac tcggccttt gttgaaaac aagtacaag gttttacaa ttatatctg gatgaagta tttgttgac aaattggta aagtatgtt gctttattg ccaatgcaa agcgattta gaccgaaca aggaatcaa tttttttgg gaaaacacc gatgagttt tggaacaat gttatagga atttgccag gatatctat aatgcaatc cccacattt gggttcaaa gttttacca aatatactg aaaagtttg ctcgtctgt ttccccgcg tttcaagac caacacaag caacatatt tcggatggg ataagggac agtgtgaac aaactcaat gaaagtttg caaatcatg attgcagag gaagaagaa aagaagaaa gcttctata caaaagaaa gtttcaaca ttaattgag gaagaacca ccggatgat tttcatcaa atttctgtc taccccaat tcacttgaa ctccttgga ggcactcga gtttttctg cgtaaaaac atagtagaa ggaccttat aaagacgta caccactac ttggacgtc cagtttaga ctgctaaga gaagacttt gttgggcct ttgagaaat gggatagcc acgtaccga acagaatca acccagaag cagcgaatt gaaaatgtc aatatttac aaaaaagta cagttttta aatcaagag acaatcaac gaacaatac tgcattcga ttaaggttc gacttttca cgcaaaaca aagaaattt cgctttgaa gactccaag aaattcatg tttggttct ctcctctgc ttcacctgc gacaacttc aagtcacta ctctttggc aaaattgct cagcgagac gtgaaagac cttgaaaag ggggaactg attgtaggt tttgatcaa aatgtgctc gtggactac aaagcgaac tatttgatg atcgaatgc ggaatctac ttcgagccg tatttccac gtcctcaac gtcctcaaa aacataaac actgacgaa ttcccgatg gaaaaatac atcattcgc gtggacccc acacctcaa ccgccggaa tacctcaac gacccaacc ccccgtaaa atctcaatc cgtggtcat gattttttc cctctcctc aactggccc aacaccaac ttctacact ttcaaccaa tcgcaacta agggccttc cgggcagct ttaacgcaa gagttctcc gttattcaa ggcccacct ggcactggg aagaccttc cttggcctc aaaatcgcc cacactttg ctgcaaaat caggccatt tggttcaag aaaaccccg atgttggtc atctgctac accaaccat gccctcgac cagttcttg gagggtttg gctctggcc actgacaga atcattcga attggggga cagagcaga aaccaaaat gtggcaaag ttcaatttg agaaatata aagttccgg acaaacccg gccgtttta caacaaaga cacggagtg aaagtcctc ttgctacaa atccagaat ataacggaa aacttgaaa gaaatcgat ttttatcac agcatcaga gatttcaaa gtttttatt gatgtgata ccgaatttt attacaaca tggtttcat cgcgctaca atagacgaa ctactggac tggttgctg cccgaatat gaaaatgaa gacttggat tcgactaat gaagttaaa agtcctagg agtgttgca gaggtccag gacaaagtc gctaacttg cagattgaa gatgaaatg tttgatgaa aatttgtcg gattatgaa atcgaagaa gatgaaaat atgaaaatt gagttggat gatcttttc gatgaagtg aatgcaatg gcggttcag tgcagcatt ccccttatt acgctctcc tcacttcag caaaggatt tctgcacta gaaagaact ttccagcaa ctgaagcaa aaggaagag ataactatg gaggatatt ttcgaagaa gatcgaatt gtcttcgaa ttgtataaa ctgaaattt cgttacgac agtttaaat tttagacta accgagggc aagcaaatg cgtttcatg aaaagaaga gttcctgat tggtcagag ttcacgaat cctcacaaa ttgtcaccc gacgatcgc tggaattta tactttttt tgtttaaat ttatacaga cagaaatta aatgagaag ctgagcgta ctaaatgag cagttccgc gaacaacac aaaatctat gaagaaatg cgggacatt gaaaacacg aaagcaatg aaaaacgtg cttgttgtt ggtatgacg accactgga gcagcccgt ctaagatct tcgttgcaa acgttgaaa agtccaatc gttattgtc gaggaagca gcggagatt ctagaagct cacatcgtt tcctctttg accaaacat tgcaaacac ttgattctc atcggagac catcagcag ttgaaaccg agcactgcg agttataac atagaaaag ttttacaat ttggggata agtctattt gagagaatg gttgtgaac agaatccag ttgaatacc ctaaacgtg caacacaga atgaggccg gagattgcc agtttggtc tcgccgacc atttatcca accctgcaa gatcacccg tctgtgaac gacagacct gatataaaa ggggtcgat aattgcttg tttttcatc gaccacaag caccctgaa gcaaattgt gaaggcaaa agcaagaag aattaccat gaagttgac ttcttgatc tatttcgca aggcatctt attctgaac ggttacgaa cccggaaac atcacaatc ctggcggct tacttgggg caaatgttc gagttgcag aaagagagg cgcaaacac aatgagctt ctagcgaac gtaagaatt gcagttttg gataactat caaggtgaa gaatgcgat attatccta ctttcgctg gttcgaaac aacgaggaa aacaaaatc gggttccta tcaattgaa aacagagtt tgtgtggcg ctttcaaga gccagaaac ggcttctat ctcatgggg aacatggac cagttgtgt gccgctagt caactttgg cgtgagatt tacaagact tttgaacgc cagaatgcc attggaccc cacctagcc ctacggtgc caagtgcac ccagataag gtcacgtat gtggcaagc ggaaaggat tttctgaat atatcagaa ggagggtgc aaccagaaa tgtggggct gatttgaat tgcggtcat cagtgcact tctctttgc cacgtgtta aatcgagat catgcaagc tacaggtgc cttgaaacg tgcgggaaa aaattgtgc gataaaaat gaactacat ttatgccaa agaaggtgt taccaagag tgtggtcct tgcacttac aaggtgtca aggacccta aaatgtggg catgttgtg aacttggag tgtcacctg gatcctgca acttacagt tgtgaggag ctcgtgcct acgcaactt ccctgtggt cacaacgct gacaagcct tgccactgc aatccggaa acttttcct tgcccgttc ccctgcgaa aaacaggtg gaaccttgt ggccatgcg tgcactaga aattgccac gttcggcag gatccagat cacttagaa tacaaatgt aagaaacgt tgcgctaaa aaccaaaag aattgcaaa gccgaaccc gaaaaacac aagtgcaag aaactatgt tttgaggat tgtgacgcc tgtatggtc cgtgttagg aaacagcga accatctgt ccacatttt tacgacgtg ccttgttcc agtaacgtg gacgatata agctgcgaa aagccttgc accaaaata ttgccttgc aatcacaaa tgcaaaaat aaatgcaat caaccgtgc ggaaactgc aaagaaaag gttgagaaa caaatcccg gtatgtgcc catacaatc aaagtaaaa tgtagcgaa gaacctctc cgaaaattc tgtaccaaa aaatgtcct ctgacacta ccctgtggc cacccttgc acgaaaaaa tgcaacgag tcttgcacc ccaacttgc accgttctg gccacctgt gcattgaaa gccccttgt ggccacacc gtgaccaag ttaccgtgc cacctagcc catggtgcc ctcgaccct aaaaccctg ctacaacac tgcaaccac ccttgcggt ggccagtta ctgtgcgag cacacctgc tcgggaacc tgcggggag tgctcgcag ggccgcatc cacaaacgc tgcgacgca aaatgcggg aaccccctg gtgtgcaac cacatctgc gagttcccc tgcagacag gcctgcagg ccctgcgtg cgcccttgc ccctacaag tgcaagcac agcgcctgt gggaagcgc tgcggcgat atctgcgtc ccttgcaag gagagctgc ccccgcagg tgcaagcac cgaaggtgc tccaagtac tgcggggaa atctgcgac gtgccccct tgtcaagag ccttgcagc aaattactg gcatgtggc caccaatgt atcgggttc tgcggcgat ccttgccct caaaaatgc cgcatttgt gacgaagac gaggtgacc gaagtgttt ttcgggttc gaggctgaa gacgatgcc cggtttgtg ctcctggag gactgcggc cacattttc gaaaatacg ggactggaa acgtggctg gggcaagac gacggggaa gtgatcaag ttcaaggag tgtccgcgg tgcaaaacg gcgatcaaa acaacggcc cgattcagt gattatgtg aagaaagcg aaggccgat gtggccaaa gttaagaaa aatctgcac gggactgag aatgagttg aacatgaag aggcaagaa ctggcgaga ctgctacta aaagttagg acgaatgca caagatgtg aaatgttcc aacttcttt aaagatatt ttgggcgcg atggaaacc cgtctgcga atttctcta atgggcaag cgccaacgt atcaacaaa tttgagttg aatgcaatt gaatccaaa ctccaaatc atcgaacac atcgttgac attggcata aactgcccc acggctctt tcaaaaaat cgctgctcc tctcaggtc aaattcata ctcaagatt ttggaacgt gaggacaac tacgtaacg gatcaagaa atagacgat atcccgtac gaattgaac cgtttgtca cgaatcgcc gaacttgaa gaaatcgaa agttcgcca aacttcctc caacgatat tcaaattct tccacagtt aggcgcgtt accgaacaa attgaaggc tttctcttc ggtttcggc aagttttat gaccaggat gaacaaacg gtcaaggaa ttattgcaa gagttgagt cgattggtc gcgtcggga ataggaatt tcagaacag gaaaaaagg gaaattgtt aaagccatg ggtttcagg caagggcat tggttcaag tgccccaat ggccaccct tactgcata gctgactgt gggggcgca atggtggag tcaaaatgc ccggaatgt gcgtgcaaa ataggggga ggtagtcac acactgcgc gccgataat gcgttggct tctgaaatg gatggagcc cagtttccc gcttggtcg gaaactgca aacaatatg ggcaatttt ttgttagat ttcgatgat tgatccagt taattaaac acgattata cttt