| General information | |||
| TC010123 | 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_007422.5: 14374359..14405187 | |
| Sequence information | |||
| Transcripts / Proteins (2) | 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 |
|
TC010123 | |
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_01652 | 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: 100.0% (includes death as pupa, adult)
eclosion not fulfilled (with pupal and adult features) - Number of animals showing the phenotype: 2 (3 dpi)adult lethality - Number of animals showing the phenotype: 2 (9 dpi)
adult lethality - Number of animals showing the phenotype: 6 (11 dpi)
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: 100.0% (includes death as larva, prepupa, pupa)
Lethalities 22 days after larval injection: 100.0% (includes death as larva, prepupa, pupa, adult)
- >TC010123 RA
MIANGQIVG DGSWNLRVL VTDLQAERQ LRVKSDLHI GGVMLRLVE DLEISMDWS DHALWWPSK NIWLTRTRS TLDQYGVQA DALLHFTPM HKNLRVQLP DLRYLDMKV DFSVKTFSA VIHLCKDLG IRHPEELSL CKPLEPCHL KYNYNSMPK KKVENGVAK NGHPNLPPD TNTFIANSS PDGSTGSLD KSGFMCAPV TPSRPPPSS TPVSSPVGN TGSWGRQVY NSETNGYSP NLTTSISSE QLNGSYDSF LAQTPPPPS PEARKHMIR PKSLVEKAR MNVAWLDSS LSIMEQGVR EYDTLCLRF KFYVFYDLN PKYDAIRIN QIYEQARWQ LLNEEIDCT EEEMLMFAA LQLQVNLQA KQPQPHLDN GNVSSPVED DIDAALTDL QTTLEGSTI NSYANDITQ VPELCDELR FLKPKRFTL KAYKRYWFT CRDLHLYLY KSKEEMIHR QNSIYDINL RGCEVTPDV NISQNKYVI KLEVPSAEG MSEMYIRCD EESQYAKWM ACCRLASKG RSLADSSYE TEKQTILEF LQMQRPADE PAINPSSLD LQAEDYIAP RFLKKFRGK NQRILEAHA NVKDLPLIE AKMNYIKAW QSLPDYGLT LFVIKFMGS KKEELLGVA PTRIMKMDI TTGDHLKTW RYNTMIAWN VNWEVKHMM IQFEDCNIM FSCSSADCK VVHEFIGGY IFLSTRSKD ASQTLNEEL FHKLTGGWS
- >TC010123 RB
MIANGQIVG DGSWNLRVL VTDLQAERQ LRVKSDLHI GGVMLRLVE DLEISMDWS DHALWWPSK NIWLTRTRS TLDQYGVQA DALLHFTPM HKNLRVQLP DLRYLDMKV DFSVKTFSA VIHLCKDLG IRHPEELSL CKPLEPCHL KYNYNSMPK KKVENGVAK NGHPNLPPD TNTFIANSS PDGSTGSLD KSGFMCAPV TPSRPPPSS TPTGSWGRQ VYNSETNGY SPNLTTSIS SEQLNGSYD SFLAQTPPP PSPEARKHM IRPKSLVEK ARMNVAWLD SSLSIMEQG VREYDTLCL RFKFYVFYD LNPKYDAIR INQIYEQAR WQLLNEEID CTEEEMLMF AALQLQVNL QAKQPQPHL DNGNVSSPV EDDIDAALT DLQTTLEGS TINSYANDI TQVPELCDE LRFLKPKRF TLKAYKRYW FTCRDLHLY LYKSKEEMI HRQNSIYDI NLRGCEVTP DVNISQNKY VIKLEVPSA EGMSEMYIR CDEESQYAK WMACCRLAS KGRSLADSS YETEKQTIL EFLQMQRPA DEPAINPSS LDLQAEDYI APRFLKKFR GKNQRILEA HANVKDLPL IEAKMNYIK AWQSLPDYG LTLFVIKFM GSKKEELLG VAPTRIMKM DITTGDHLK TWRYNTMIA WNVNWEVKH MMIQFEDCN IMFSCSSAD CKVVHEFIG GYIFLSTRS KDASQTLNE ELFHKLTGG WS
- >TC010123 RA
atgatagcg aacggacag atagtgggt gacggctcc tggaacctg cgggtgcta gtcaccgat ttgcaagcg gagcgccaa ctccgagtg aagagcgac ttacacatc ggaggagtt atgctgaga ctcgtcgag gacttggaa atctcaatg gactggtcg gaccatgcc ctgtggtgg cccagcaag aacatttgg ttgacgcga acccgctca accctcgac caatacggc gtccaagcc gatgccctt ttgcacttc acccccatg cacaaaaac ttgcgcgtc caactgccc gacttgcgc tacttggac atgaaagtg gatttttcc gtaaaaaca ttttcggcc gttattcac ctgtgtaaa gacctgggt attaggcat ccggaagag ttgtcgttg tgcaaaccg ctcgaacct tgccattta aaatacaat tacaactcg atgccaaag aagaaagtt gaaaacggg gttgcgaaa aacgggcat cccaacctt ccgccagac accaacacc ttcatcgcc aacagcagt cccgatggg agcaccggg agcttggac aagagtggg tttatgtgc gctcctgtc acaccctcg aggcctccg ccctcctcc acaccggtc agcagtcct gtgggaaat acaggatcg tggggtcgt caggtgtac aattcggag actaacggt tattccccc aatttgacg acaagtatc agttcggag caattgaat ggcagttac gacagtttt cttgcccaa actccacct ccgccgagt cctgaggct cggaaacat atgattagg ccgaaatcg cttgtagaa aaagcgaga atgaatgtg gcttggtta gattcgtct ttgtcaatt atggaacaa ggagtcaga gaatacgat acgttatgt ttgaggttt aaattttac gtgttttat gacctaaac cccaaatac gatgcgatt cgaatcaat cagatttat gaacaagcg aggtggcaa ttgttaaat gaagagatt gattgtacg gaagaagaa atgttaatg tttgccgcc ttacagttg caagttaat ttacaagct aaacaaccc caacctcat ttggataat ggcaatgtg tcttcccct gttgaagac gacatcgac gcggcctta accgacctt caaacaact ctcgaggga tcaacaata aattcctac gccaacgac atcacacaa gtgcctgaa ttatgcgac gaattacgg tttttaaaa ccgaaaagg ttcacatta aaagcgtac aaaagatac tggttcaca tgccgcgac ctacatctc tacctgtac aaatccaaa gaggaaatg atccatcga caaaactcg atctacgat atcaatttg cgcggctgc gaggtcact ccagacgtc aacattagt caaaataaa tatgtaatc aagttggaa gtaccgagc gccgaaggc atgtctgaa atgtacata cgatgtgat gaagaaagt cagtacgca aaatggatg gcctgctgt cgattggct tcaaaaggc cgatcgctg gccgacagc agttacgaa acggagaaa caaacaatt ttggaattt ttgcaaatg caaaggcca gcggatgag cctgccatt aatcctagc agtctcgat cttcaagca gaggattac atcgcaccg cgatttttg aagaaattt cgagggaaa aatcaaaga attttggaa gcacacgcc aatgttaaa gaccttccg ttgattgag gccaaaatg aactacatc aaggcgtgg cagtcgctc cctgactat ggtctaact ctatttgtt atcaaattt atgggaagc aaaaaagag gagttgcta ggggtggcc ccaacgcga atcatgaaa atggacata acgacaggt gatcacttg aagacatgg aggtacaac accatgatc gcctggaat gttaactgg gaagtgaaa catatgatg atacagttc gaagattgt aacatcatg ttcagttgc tcctcagct gattgtaaa gtcgttcac gaatttatc ggcggttat attttcttg tcaacgcgg tcgaaggat gcgagtcag acgctaaac gaagaactc ttccacaaa ctgacaggt ggctggtcc taa
- >TC010123 RB
atgatagcg aacggacag atagtgggt gacggctcc tggaacctg cgggtgcta gtcaccgat ttgcaagcg gagcgccaa ctccgagtg aagagcgac ttacacatc ggaggagtt atgctgaga ctcgtcgag gacttggaa atctcaatg gactggtcg gaccatgcc ctgtggtgg cccagcaag aacatttgg ttgacgcga acccgctca accctcgac caatacggc gtccaagcc gatgccctt ttgcacttc acccccatg cacaaaaac ttgcgcgtc caactgccc gacttgcgc tacttggac atgaaagtg gatttttcc gtaaaaaca ttttcggcc gttattcac ctgtgtaaa gacctgggt attaggcat ccggaagag ttgtcgttg tgcaaaccg ctcgaacct tgccattta aaatacaat tacaactcg atgccaaag aagaaagtt gaaaacggg gttgcgaaa aacgggcat cccaacctt ccgccagac accaacacc ttcatcgcc aacagcagt cccgatggg agcaccggg agcttggac aagagtggg tttatgtgc gctcctgtc acaccctcg aggcctccg ccctcctcc acaccgaca ggatcgtgg ggtcgtcag gtgtacaat tcggagact aacggttat tcccccaat ttgacgaca agtatcagt tcggagcaa ttgaatggc agttacgac agttttctt gcccaaact ccacctccg ccgagtcct gaggctcgg aaacatatg attaggccg aaatcgctt gtagaaaaa gcgagaatg aatgtggct tggttagat tcgtctttg tcaattatg gaacaagga gtcagagaa tacgatacg ttatgtttg aggtttaaa ttttacgtg ttttatgac ctaaacccc aaatacgat gcgattcga atcaatcag atttatgaa caagcgagg tggcaattg ttaaatgaa gagattgat tgtacggaa gaagaaatg ttaatgttt gccgcctta cagttgcaa gttaattta caagctaaa caaccccaa cctcatttg gataatggc aatgtgtct tcccctgtt gaagacgac atcgacgcg gccttaacc gaccttcaa acaactctc gagggatca acaataaat tcctacgcc aacgacatc acacaagtg cctgaatta tgcgacgaa ttacggttt ttaaaaccg aaaaggttc acattaaaa gcgtacaaa agatactgg ttcacatgc cgcgaccta catctctac ctgtacaaa tccaaagag gaaatgatc catcgacaa aactcgatc tacgatatc aatttgcgc ggctgcgag gtcactcca gacgtcaac attagtcaa aataaatat gtaatcaag ttggaagta ccgagcgcc gaaggcatg tctgaaatg tacatacga tgtgatgaa gaaagtcag tacgcaaaa tggatggcc tgctgtcga ttggcttca aaaggccga tcgctggcc gacagcagt tacgaaacg gagaaacaa acaattttg gaatttttg caaatgcaa aggccagcg gatgagcct gccattaat cctagcagt ctcgatctt caagcagag gattacatc gcaccgcga tttttgaag aaatttcga gggaaaaat caaagaatt ttggaagca cacgccaat gttaaagac cttccgttg attgaggcc aaaatgaac tacatcaag gcgtggcag tcgctccct gactatggt ctaactcta tttgttatc aaatttatg ggaagcaaa aaagaggag ttgctaggg gtggcccca acgcgaatc atgaaaatg gacataacg acaggtgat cacttgaag acatggagg tacaacacc atgatcgcc tggaatgtt aactgggaa gtgaaacat atgatgata cagttcgaa gattgtaac atcatgttc agttgctcc tcagctgat tgtaaagtc gttcacgaa tttatcggc ggttatatt ttcttgtca acgcggtcg aaggatgcg agtcagacg ctaaacgaa gaactcttc cacaaactg acaggtggc tggtcctaa
- >TC010123 RA
ggacagacg attggtgac actgttcca tgactttct ccctccact tttacaaga gataatgat aaacacacg aagcaccga ccaatcacg cgtagttag ttgttctta gaaggacta agtcacgag acgattcgg gtttgcaat gagccccgc gttgtaaac aatgtcgga ataaataat tagagtttt attatctta tcttaatga tagtgaagt gacaagtaa ttgataata atgatagcg aacggacag atagtgggt gacggctcc tggaacctg cgggtgcta gtcaccgat ttgcaagcg gagcgccaa ctccgagtg aagagcgac ttacacatc ggaggagtt atgctgaga ctcgtcgag gacttggaa atctcaatg gactggtcg gaccatgcc ctgtggtgg cccagcaag aacatttgg ttgacgcga acccgctca accctcgac caatacggc gtccaagcc gatgccctt ttgcacttc acccccatg cacaaaaac ttgcgcgtc caactgccc gacttgcgc tacttggac atgaaagtg gatttttcc gtaaaaaca ttttcggcc gttattcac ctgtgtaaa gacctgggt attaggcat ccggaagag ttgtcgttg tgcaaaccg ctcgaacct tgccattta aaatacaat tacaactcg atgccaaag aagaaagtt gaaaacggg gttgcgaaa aacgggcat cccaacctt ccgccagac accaacacc ttcatcgcc aacagcagt cccgatggg agcaccggg agcttggac aagagtggg tttatgtgc gctcctgtc acaccctcg aggcctccg ccctcctcc acaccggtc agcagtcct gtgggaaat acaggatcg tggggtcgt caggtgtac aattcggag actaacggt tattccccc aatttgacg acaagtatc agttcggag caattgaat ggcagttac gacagtttt cttgcccaa actccacct ccgccgagt cctgaggct cggaaacat atgattagg ccgaaatcg cttgtagaa aaagcgaga atgaatgtg gcttggtta gattcgtct ttgtcaatt atggaacaa ggagtcaga gaatacgat acgttatgt ttgaggttt aaattttac gtgttttat gacctaaac cccaaatac gatgcgatt cgaatcaat cagatttat gaacaagcg aggtggcaa ttgttaaat gaagagatt gattgtacg gaagaagaa atgttaatg tttgccgcc ttacagttg caagttaat ttacaagct aaacaaccc caacctcat ttggataat ggcaatgtg tcttcccct gttgaagac gacatcgac gcggcctta accgacctt caaacaact ctcgaggga tcaacaata aattcctac gccaacgac atcacacaa gtgcctgaa ttatgcgac gaattacgg tttttaaaa ccgaaaagg ttcacatta aaagcgtac aaaagatac tggttcaca tgccgcgac ctacatctc tacctgtac aaatccaaa gaggaaatg atccatcga caaaactcg atctacgat atcaatttg cgcggctgc gaggtcact ccagacgtc aacattagt caaaataaa tatgtaatc aagttggaa gtaccgagc gccgaaggc atgtctgaa atgtacata cgatgtgat gaagaaagt cagtacgca aaatggatg gcctgctgt cgattggct tcaaaaggc cgatcgctg gccgacagc agttacgaa acggagaaa caaacaatt ttggaattt ttgcaaatg caaaggcca gcggatgag cctgccatt aatcctagc agtctcgat cttcaagca gaggattac atcgcaccg cgatttttg aagaaattt cgagggaaa aatcaaaga attttggaa gcacacgcc aatgttaaa gaccttccg ttgattgag gccaaaatg aactacatc aaggcgtgg cagtcgctc cctgactat ggtctaact ctatttgtt atcaaattt atgggaagc aaaaaagag gagttgcta ggggtggcc ccaacgcga atcatgaaa atggacata acgacaggt gatcacttg aagacatgg aggtacaac accatgatc gcctggaat gttaactgg gaagtgaaa catatgatg atacagttc gaagattgt aacatcatg ttcagttgc tcctcagct gattgtaaa gtcgttcac gaatttatc ggcggttat attttcttg tcaacgcgg tcgaaggat gcgagtcag acgctaaac gaagaactc ttccacaaa ctgacaggt ggctggtcc taagagtag tgccttaat tattattaa cgtgtcaat taaaaaaca aggaatttt gttgtagca acttctaat ccggttatt tgttagcgt tgattcagt atttggcga cattgaagt gtctatggt aaagttgta gcccgttga acctctcca attattgtt attcaaatt ttataaata tttttcaaa taatgtggc aatatctaa tagcaggtt gcgcgaaag cagtttttc ttgtaactt tactaattt actagtcta atgtcacat tttagtgac cgcctaaaa acttttggt aattctttc acctctcta tcaacccga ctagcaaaa gtgcttata ttttaaaac ataatcgtg cgctctttt gcactaaat taaaacgaa acaacagag gtaagtaaa atatttgaa atgaagaga aatgtgata acaatttaa cggaaataa ctaaatatg gtaaagggg gttttagga aacaaaata ttcactaaa taaaatgat tccttagat aaaccccga tattcaaaa atcgatgtg tccgtggta attacttta ccatcaata aagtggaca gatgctttt caactcatc tgtaactat ttattatgt aaaggtcca accaaacga ggggcctat ttcaccaac gaaattttc aatttttta catggtaac atacaattt ggtgataaa gaaattttt ccaagatgt caatgtcgc ttttcacct ttcaaacgt tttgtaaag tgtaaacag actaagaag aaagattta taaataaga acaaatttt ggttaaaca tttcaagtt aattctaac aattaaaaa aattgttaa attataatc actaaaaag atctgaaac agttactac acattgtaa aattaatta tttgaaatt ttacgcatg ttttatttt gtcagtcaa cttctttct gtgaaaaag aagactgac gaattctga gaaaaaggt ttacttcgt atagacaaa aaaacattg tgaaaaact gaaaagtaa ataaaattg tatttcttt aataattat acttttgaa acagaaaca ctctttact ttgcagaat ctgcaaagt ggaaaagca tagcgtaga agtttgttg gtgaaatgg acctctgat tttattaca gttctcagt ttttttgtt tttaacttg ttatcagga aacagataa aaaacgatc aataaaagt ttaaataaa acgctacat tcaacacta ctaacaggt gtttcttta aaaatacat aagtttgca tttttgtat atatttttt taatacgca tgcttggac attctgtat agaggcacc aatttacct acctcaggt gaaattgtc tagattcgg tttggaatt ttaacgcat tattatgca tttttaaaa aggctaaaa attttatta aacagtact gtacaat
- >TC010123 RB
ggacagacg attggtgac actgttcca tgactttct ccctccact tttacaaga gataatgat aaacacacg aagcaccga ccaatcacg cgtagttag ttgttctta gaaggacta agtcacgag acgattcgg gtttgcaat gagccccgc gttgtaaac aatgtcgga ataaataat tagagtttt attatctta tcttaatga tagtgaagt gacaagtaa ttgataata atgatagcg aacggacag atagtgggt gacggctcc tggaacctg cgggtgcta gtcaccgat ttgcaagcg gagcgccaa ctccgagtg aagagcgac ttacacatc ggaggagtt atgctgaga ctcgtcgag gacttggaa atctcaatg gactggtcg gaccatgcc ctgtggtgg cccagcaag aacatttgg ttgacgcga acccgctca accctcgac caatacggc gtccaagcc gatgccctt ttgcacttc acccccatg cacaaaaac ttgcgcgtc caactgccc gacttgcgc tacttggac atgaaagtg gatttttcc gtaaaaaca ttttcggcc gttattcac ctgtgtaaa gacctgggt attaggcat ccggaagag ttgtcgttg tgcaaaccg ctcgaacct tgccattta aaatacaat tacaactcg atgccaaag aagaaagtt gaaaacggg gttgcgaaa aacgggcat cccaacctt ccgccagac accaacacc ttcatcgcc aacagcagt cccgatggg agcaccggg agcttggac aagagtggg tttatgtgc gctcctgtc acaccctcg aggcctccg ccctcctcc acaccgaca ggatcgtgg ggtcgtcag gtgtacaat tcggagact aacggttat tcccccaat ttgacgaca agtatcagt tcggagcaa ttgaatggc agttacgac agttttctt gcccaaact ccacctccg ccgagtcct gaggctcgg aaacatatg attaggccg aaatcgctt gtagaaaaa gcgagaatg aatgtggct tggttagat tcgtctttg tcaattatg gaacaagga gtcagagaa tacgatacg ttatgtttg aggtttaaa ttttacgtg ttttatgac ctaaacccc aaatacgat gcgattcga atcaatcag atttatgaa caagcgagg tggcaattg ttaaatgaa gagattgat tgtacggaa gaagaaatg ttaatgttt gccgcctta cagttgcaa gttaattta caagctaaa caaccccaa cctcatttg gataatggc aatgtgtct tcccctgtt gaagacgac atcgacgcg gccttaacc gaccttcaa acaactctc gagggatca acaataaat tcctacgcc aacgacatc acacaagtg cctgaatta tgcgacgaa ttacggttt ttaaaaccg aaaaggttc acattaaaa gcgtacaaa agatactgg ttcacatgc cgcgaccta catctctac ctgtacaaa tccaaagag gaaatgatc catcgacaa aactcgatc tacgatatc aatttgcgc ggctgcgag gtcactcca gacgtcaac attagtcaa aataaatat gtaatcaag ttggaagta ccgagcgcc gaaggcatg tctgaaatg tacatacga tgtgatgaa gaaagtcag tacgcaaaa tggatggcc tgctgtcga ttggcttca aaaggccga tcgctggcc gacagcagt tacgaaacg gagaaacaa acaattttg gaatttttg caaatgcaa aggccagcg gatgagcct gccattaat cctagcagt ctcgatctt caagcagag gattacatc gcaccgcga tttttgaag aaatttcga gggaaaaat caaagaatt ttggaagca cacgccaat gttaaagac cttccgttg attgaggcc aaaatgaac tacatcaag gcgtggcag tcgctccct gactatggt ctaactcta tttgttatc aaatttatg ggaagcaaa aaagaggag ttgctaggg gtggcccca acgcgaatc atgaaaatg gacataacg acaggtgat cacttgaag acatggagg tacaacacc atgatcgcc tggaatgtt aactgggaa gtgaaacat atgatgata cagttcgaa gattgtaac atcatgttc agttgctcc tcagctgat tgtaaagtc gttcacgaa tttatcggc ggttatatt ttcttgtca acgcggtcg aaggatgcg agtcagacg ctaaacgaa gaactcttc cacaaactg acaggtggc tggtcctaa gagtagtgc cttaattat tattaacgt gtcaattaa aaaacaagg aattttgtt gtagcaact tctaatccg gttatttgt tagcgttga ttcagtatt tggcgacat tgaagtgtc tatggtaaa gttgtagcc cgttgaacc tctccaatt attgttatt caaatttta taaatattt ttcaaataa tgtggcaat atctaatag caggttgcg cgaaagcag tttttcttg taactttac taatttact agtctaatg tcacatttt agtgaccgc ctaaaaact tttggtaat tctttcacc tctctatca acccgacta gcaaaagtg cttatattt taaaacata atcgtgcgc tcttttgca ctaaattaa aacgaaaca acagaggta agtaaaata tttgaaatg aagagaaat gtgataaca atttaacgg aaataacta aatatggta aagggggtt ttaggaaac aaaatattc actaaataa aatgattcc ttagataaa ccccgatat tcaaaaatc gatgtgtcc gtggtaatt actttacca tcaataaag tggacagat gcttttcaa ctcatctgt aactattta ttatgtaaa ggtccaacc aaacgaggg gcctatttc accaacgaa attttcaat tttttacat ggtaacata caatttggt gataaagaa atttttcca agatgtcaa tgtcgcttt tcacctttc aaacgtttt gtaaagtgt aaacagact aagaagaaa gatttataa ataagaaca aattttggt taaacattt caagttaat tctaacaat taaaaaaat tgttaaatt ataatcact aaaaagatc tgaaacagt tactacaca ttgtaaaat taattattt gaaatttta cgcatgttt tattttgtc agtcaactt ctttctgtg aaaaagaag actgacgaa ttctgagaa aaaggttta cttcgtata gacaaaaaa acattgtga aaaactgaa aagtaaata aaattgtat ttctttaat aattatact tttgaaaca gaaacactc tttactttg cagaatctg caaagtgga aaagcatag cgtagaagt ttgttggtg aaatggacc tctgatttt attacagtt ctcagtttt tttgttttt aacttgtta tcaggaaac agataaaaa acgatcaat aaaagttta aataaaacg ctacattca acactacta acaggtgtt tctttaaaa atacataag tttgcattt ttgtatata tttttttaa tacgcatgc ttggacatt ctgtataga ggcaccaat ttacctacc tcaggtgaa attgtctag attcggttt ggaatttta acgcattat tatgcattt ttaaaaagg ctaaaaatt ttattaaac agtactgta caat
