| General information | |||
| TC004742 | 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_007416.3: 6296636..6302543 | |
| 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 |
|
TC004742 | |
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_00761 | 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)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: 40.0% (includes death as larva, prepupa, pupa)
Lethalities 22 days after larval injection: 40.0% (includes death as larva, prepupa, pupa, adult)
remarks: later than schedule,
- >TC004742 RA
MSSSTPFWR KIHQTISGH SQNITEDIV AEILKLSKT ELLEGLLSY RPYTKKGYH DWVAVSDVS ICMKEFVHD ISQVLNLDT ITAYPVMCN YLMFEYYGK IKEFWQITR MGSTLHLKE SIWNFYTAE RMFLLKTWR HIFECVNKD YEFAEQYKD FLKTIKIEE LQNNLITQF EELLNEVTS NNFDERSPY NENWLCRNI REQIEILLI LILTATHKP LATAKLSHL IKLFTQNNF GTQPAFFDI RTYAKPQDL DQLMFAEVG CYFAILDNY WDKEIPDFW KQDSTSELD KSLLNLHLN PGHIIIMLA WLTLKFKSL NRDQDAEKI LELFKRQPF ILWHKILSD SMFKNCTVG EVVLRATHK MLDELCLMF DDRKILYED GGVIKILAE FLKQPDLAE ICLQTRNGL DCMTEIAIE SFPHDFYSF TTIAQSLLG LRDQYKQTM QLLNNFPSF LSEMTWTIP KEQISVLRK NQPAFPDSD LIVFPPKTV VRSYRIFGK NLIQYKGKY SYFFHLEYL VTTLINNGL CHRKFDEAL VQKVIAGYG VVIQAIRYC VDMDLRTVG LERIILKLN KLLVHLDGP LLHFELVKL YFDLQNALM LYRQSSFFK ICSEIVWKI FFPKLKPEY LTQDVKILL LNQYVFEDR IFMQLFQKE EFKEDHTLL LTYIKILSN IIKVKFQFK NFQLSGVVY LFNYVFLQH QTWVYKDEN QKTQITLEC LKIFHYVLQ KQVEDLTDI ELKIFQLCN HAFLHNVYV IDSFLRLFV KERYYLIYA MERESNWHQ GPSLDTLQC IRMQLALLL LIMNRKKAI GDCCFSERL PFVVKPVAS YFTNAYSPL IAELSCRFL EKLSQDPFV PLLALLELD HYQVQSLFL ERLRDPLEE ENVKMAIID LINTCISSQ DGMTAAFFN LKCFMYWDG AENDVINGD SVSDFMVDY LQNIKKSHE YFKNPLQLG ILRLLFNLW LNHRENLIE NIASLKDFW PVMADPFFC DYKQDIEIY TIILRIINL EIGANIDKV DEKLVKTID KFLKDKKRI AQWNKFVLA SSQSSAKNK LDLLSAWME FLVLTKKVI PDSFENDVK FALVACCLD GLHPPEGGF SNIETIYLW SQLYLLLIS TWSVYEKKE QVVLGKLQT FLHALNTYY KYLTPKIKE NILCAVNRT ILDLQEYFS VNTAQLLTF LYSVGPLID AEYYYLVSE AFETEDDAE KVRRLKPWL IIVFIGNSI MALDRCEEI GLWFYYEQF LQRVMDSVG SMMEHSATL PFAKLAIDF LITYAESPF VKDFLKEDL FSFYFKVRP PPLTISVGE AILSGLPVN LKEWWFILI TLIKLNRIL IQQIGDAML RTCFTFITQ HDLLLKEIV SLTKYTVDM SALTLVCET LKLILVVLT QMPHWRVDS PYSYNLIME GIKTTIHAC VLSILGYKK INFYDIING KNLNIIDVT PTDLLVSVL NKLTEITYW GCSCLKKTK TDLVTQLNA HYETENLAL LLVENDFSV PRFELPITP RLTYGSLLC LADYLCKTL NQLSPATPS RISRSRQVS LTDLNNQMK VKSLGKIFE HCTATYEEF IGFMVYIQK SDDLPNEKF SSFLHYDMT EFSGTTSAV SQLDYHLVK NALEALMSF LAGEIFFAI RNLRGEAVF AYRRELSSE IQFFYEFVR KRTSEQYSA MVATPRSTS SIAQTEPDL IKRYIITKV MDNDTSDIV DNNFVLVIS QWLIKFCHL SN
- >TC004742 RA
atgtcttct tccacccct ttttggaga aaaatccac caaacaata tcgggccat agccaaaac attactgaa gacatagtc gcggaaatt ttaaaactc tcgaaaaca gaattactt gaaggcctc ttgagctat cgaccgtac acaaaaaag ggttatcat gactgggtt gcggtctcg gacgttagc atttgcatg aaagaattc gttcatgat ataagccaa gttcttaat ctggacaca ataacggct tatccggtc atgtgcaat tacctgatg tttgagtat tatggaaaa attaaggag ttttggcaa ataactaga atgggttca accttacat ttgaaagag agtatttgg aatttttac acggctgaa aggatgttt ttgctgaag acgtggaga cacatattt gagtgtgtc aataaggat tatgagttt gctgagcag tacaaggat tttctaaaa acgatcaaa atcgaggaa ttgcaaaat aacttgatt acgcagttt gaggagctc cttaacgag gtcacgtct aataatttt gatgagagg agtccgtac aatgaaaac tggctgtgt cggaacatc agggaacaa atagaaatt ctcctgatt ctcatccta acagccacc cataaaccc ctggcaact gcaaaactc tctcatttg attaagtta ttcactcag aataatttc ggcacccag ccggcattt ttcgacatc cggacgtac gcaaaaccg caagacttg gaccagtta atgtttgcc gaagtcggg tgttacttt gccattttg gacaattat tgggataag gaaattcca gatttttgg aaacaagac tcaacaagc gagttggac aagtcgctg cttaactta cacctcaac ccagggcac attataata atgctggcc tggttaact ctgaaattt aaatcgcta aatcgtgac caagacgcc gaaaaaata ctagagctg ttcaaaaga cagccgttt attttgtgg cacaaaatc ctctcagat agtatgttt aagaactgt acggttgga gaagtcgtt ttgcgagca acgcataaa atgctcgac gagctttgt ttgatgttc gacgaccga aaaattctc tacgaggac ggcggcgtg attaaaatc cttgccgag tttttgaaa caaccagac ttggccgaa atttgtttg caaacgcga aatggttta gactgcatg acggaaatc gcgattgag tcatttcct cacgatttt tattcgttc accaccatt gcacaatcg ctgctaggc ctgcgtgac caatacaaa caaaccatg caattgttg aataatttc ccgtcgttt ttgtccgaa atgacctgg accattcca aaggaacaa atctcggtt ttgcgcaaa aaccagccg gcgtttccc gacagcgac ctgatagtc ttcccgccg aaaacggtg gtgcggtcg taccgaatt ttcggaaaa aacctgata cagtacaag gggaagtac agctacttc ttccacctc gagtatttg gtcacaacg ctgataaac aacgggctg tgccaccgc aagtttgac gaggctttg gtccagaag gtcattgcc gggtatggc gttgtgatc caagctatt aggtattgc gtggacatg gacttgaga acagtgggg cttgagaga attattctg aaattaaac aaactccta gtgcatttg gacggccct ttgctgcac tttgagttg gtcaaactt tacttcgac ttgcaaaat gcgctaatg ctgtaccga caaagcagc ttcttcaag atctgctcc gaaatcgta tggaaaata tttttcccg aagctcaaa ccggaatat ttgacccaa gacgttaaa attttgctg ctgaatcag tacgtgttt gaggaccga attttcatg caattgttc caaaaagag gagtttaag gaagatcac actctcctt cttacctac atcaagatt ctaagcaac attattaag gtgaaattc caattcaag aatttccaa ctctcgggc gttgtttat ttatttaat tacgtcttt ttgcaacac caaacttgg gtctataag gacgagaat caaaagacg caaatcaca ctcgagtgc cttaaaatt tttcattac gttttgcaa aaacaggtc gaggatttg accgacatt gagctgaaa atcttccag ttgtgcaat cacgctttt ttgcacaac gtctacgta atcgactcg ttcctgcgc ttgtttgtt aaggagaga tattatttg atttacgca atggaacgg gaatcgaat tggcatcag ggcccgtct ttagacacc ttacaatgt atcaggatg cagttagct cttctcctg ttaattatg aatcggaaa aaagcaatt ggtgattgt tgcttcagt gagagactg cccttcgtc gtaaaacca gtagcttcg tattttacc aacgcgtat agcccgctt attgccgag ctgtcctgc cgttttttg gaaaaatta tcacaagat ccgtttgtt cccttactt gcgttactt gagctggac cattatcaa gtccagtcg ctgtttttg gaacggtta agggaccct ctggaggag gaaaacgtg aaaatggca atcatcgac ctaattaac acgtgtatt tcaagccag gacggcatg actgcggcc ttcttcaac ttgaagtgt ttcatgtac tgggacggg gccgagaac gatgttatt aacggcgac agcgtgagc gactttatg gtcgattac ttgcaaaat attaaaaag tcgcacgaa tatttcaaa aatccgctc cagctcgga atcctccga ctcctcttc aacttgtgg ctaaaccac cgggaaaat ttaatcgaa aacatcgca agcttgaag gatttctgg cccgtgatg gcggacccg tttttctgc gactataag caagacatt gaaatttac acaatcatc ctccgcata attaacttg gaaatcggg gctaacata gacaaggtg gacgaaaaa ctcgtaaaa accatcgat aagtttttg aaggacaag aaacgcatt gcacagtgg aacaaattc gtgcttgcg tcctctcaa agcagtgcg aaaaacaag ctggacttg ctcagtgcc tggatggag tttttggtt ttgacgaag aaagttata cctgatagt ttcgagaac gacgttaag ttcgccctc gttgcctgt tgtctggac ggcttgcac cctcctgaa ggcgggttc agtaacatc gaaacgatt tatctgtgg tcacagctg tacttgctg ttaataagt acgtggtca gtctacgag aaaaaggag caagtggtt ttggggaag ctacaaacg ttcctgcac gcgcttaac acgtattat aaatatctg actccgaaa atcaaggag aatatttta tgtgcggtg aatcggact atcctcgat ctgcaggag tacttcagt gttaataca gcgcagttg ttgacattt ttgtactcc gttgggccg ctgattgac gcggaatat tactacctg gtgtccgaa gcgttcgag actgaggac gatgcggaa aaagtgcgg cgacttaag ccgtggttg attattgtt tttatcgga aactcgata atggcgctc gatcgctgc gaggaaatt gggttgtgg ttctactac gagcagttt ttgcaacgg gtgatggac tctgtgggg tcaatgatg gaacatagc gccactttg ccgtttgca aagctagct attgacttt cttatcact tatgcggag tctccgttc gttaaggac ttcctaaag gaggatctg ttcagtttc tactttaaa gtgcgaccg ccaccgctc acaataagt gtgggggag gcaattcta tcgggactt ccggtcaat ttgaaggag tggtggttt attttaatc actttgata aagctgaac cgtattttg atacaacag atcggcgat gcaatgcta aggacgtgc ttcacgttc atcacacaa cacgacttg ttgctcaag gaaattgtt tctttgacc aaatacacc gttgatatg tcggccttg actctcgtt tgtgaaact ttgaaactt attctggtt gttctgaca caaatgcca cattggcga gtcgactcg ccgtactct tacaactta ataatggag ggtataaaa acgacaata cacgcctgc gttttgtca atcctgggg tacaaaaaa attaatttt tatgacatc atcaacgga aaaaatctg aacattatt gacgtcacc ccaaccgac ttactagtc tcagtgttg aataaacta acagaaatt acgtactgg ggctgcagc tgtttgaag aaaaccaag accgactta gtaacccag ctaaacgcc cactacgag actgaaaac ctcgcatta ttattggtc gaaaacgac tttagtgtt ccccgattt gagttgccg atcactccg agactgacg tacgggtct ctattgtgc ctggctgac tatttgtgc aaaaccctc aatcagttg tctcctgca actccttcg agaatttcg agaagtcgt caagtgtct ctaaccgat cttaacaat caaatgaaa gtcaagagt ttggggaaa attttcgag cattgcacc gcaacgtat gaggaattt ataggtttt atggtctac atccaaaaa agcgacgat cttccgaat gaaaagttc agttcgttt ttgcactac gacatgacg gaattttcc gggacgact tcggcggtt tcccagttg gattatcat ttggttaag aacgcgcta gaggcgttg atgagtttc ctggctggg gaaatcttc ttcgcaatt cggaatctc cgaggcgaa gctgtattt gcatacagg agggagttg agttcggaa atacagttc ttctacgag tttgtccga aagcggact agtgaacaa tacagtgcg atggtggca acacctcga tcaacgtcc tccatcgca cagactgag cccgacctt atcaaaaga tacatcatc accaaagtc atggataac gataccagc gatattgtt gataataat tttgtactt gttatttct cagtggtta atcaaattc tgccacttg tcaaattaa
- >TC004742 RA
taaaagtgg cggtctata tctacataa cctgcaaac ttttgcttg tgcttgtgt gtgtatttg taatgttaa tttcccaat taaaaaaat gtcttcttc caccccttt ttggagaaa aatccacca aacaatatc gggccatag ccaaaacat tactgaaga catagtcgc ggaaatttt aaaactctc gaaaacaga attacttga aggcctctt gagctatcg accgtacac aaaaaaggg ttatcatga ctgggttgc ggtctcgga cgttagcat ttgcatgaa agaattcgt tcatgatat aagccaagt tcttaatct ggacacaat aacggctta tccggtcat gtgcaatta cctgatgtt tgagtatta tggaaaaat taaggagtt ttggcaaat aactagaat gggttcaac cttacattt gaaagagag tatttggaa tttttacac ggctgaaag gatgttttt gctgaagac gtggagaca catatttga gtgtgtcaa taaggatta tgagtttgc tgagcagta caaggattt tctaaaaac gatcaaaat cgaggaatt gcaaaataa cttgattac gcagtttga ggagctcct taacgaggt cacgtctaa taattttga tgagaggag tccgtacaa tgaaaactg gctgtgtcg gaacatcag ggaacaaat agaaattct cctgattct catcctaac agccaccca taaacccct ggcaactgc aaaactctc tcatttgat taagttatt cactcagaa taatttcgg cacccagcc ggcattttt cgacatccg gacgtacgc aaaaccgca agacttgga ccagttaat gtttgccga agtcgggtg ttactttgc cattttgga caattattg ggataagga aattccaga tttttggaa acaagactc aacaagcga gttggacaa gtcgctgct taacttaca cctcaaccc agggcacat tataataat gctggcctg gttaactct gaaatttaa atcgctaaa tcgtgacca agacgccga aaaaatact agagctgtt caaaagaca gccgtttat tttgtggca caaaatcct ctcagatag tatgtttaa gaactgtac ggttggaga agtcgtttt gcgagcaac gcataaaat gctcgacga gctttgttt gatgttcga cgaccgaaa aattctcta cgaggacgg cggcgtgat taaaatcct tgccgagtt tttgaaaca accagactt ggccgaaat ttgtttgca aacgcgaaa tggtttaga ctgcatgac ggaaatcgc gattgagtc atttcctca cgattttta ttcgttcac caccattgc acaatcgct gctaggcct gcgtgacca atacaaaca aaccatgca attgttgaa taatttccc gtcgttttt gtccgaaat gacctggac cattccaaa ggaacaaat ctcggtttt gcgcaaaaa ccagccggc gtttcccga cagcgacct gatagtctt cccgccgaa aacggtggt gcggtcgta ccgaatttt cggaaaaaa cctgataca gtacaaggg gaagtacag ctacttctt ccacctcga gtatttggt cacaacgct gataaacaa cgggctgtg ccaccgcaa gtttgacga ggctttggt ccagaaggt cattgccgg gtatggcgt tgtgatcca agctattag gtattgcgt ggacatgga cttgagaac agtggggct tgagagaat tattctgaa attaaacaa actcctagt gcatttgga cggcccttt gctgcactt tgagttggt caaacttta cttcgactt gcaaaatgc gctaatgct gtaccgaca aagcagctt cttcaagat ctgctccga aatcgtatg gaaaatatt tttcccgaa gctcaaacc ggaatattt gacccaaga cgttaaaat tttgctgct gaatcagta cgtgtttga ggaccgaat tttcatgca attgttcca aaaagagga gtttaagga agatcacac tctccttct tacctacat caagattct aagcaacat tattaaggt gaaattcca attcaagaa tttccaact ctcgggcgt tgtttattt atttaatta cgtcttttt gcaacacca aacttgggt ctataagga cgagaatca aaagacgca aatcacact cgagtgcct taaaatttt tcattacgt tttgcaaaa acaggtcga ggatttgac cgacattga gctgaaaat cttccagtt gtgcaatca cgctttttt gcacaacgt ctacgtaat cgactcgtt cctgcgctt gtttgttaa ggagagata ttatttgat ttacgcaat ggaacggga atcgaattg gcatcaggg cccgtcttt agacacctt acaatgtat caggatgca gttagctct tctcctgtt aattatgaa tcggaaaaa agcaattgg tgattgttg cttcagtga gagactgcc cttcgtcgt aaaaccagt agcttcgta ttttaccaa cgcgtatag cccgcttat tgccgagct gtcctgccg ttttttgga aaaattatc acaagatcc gtttgttcc cttacttgc gttacttga gctggacca ttatcaagt ccagtcgct gtttttgga acggttaag ggaccctct ggaggagga aaacgtgaa aatggcaat catcgacct aattaacac gtgtatttc aagccagga cggcatgac tgcggcctt cttcaactt gaagtgttt catgtactg ggacggggc cgagaacga tgttattaa cggcgacag cgtgagcga ctttatggt cgattactt gcaaaatat taaaaagtc gcacgaata tttcaaaaa tccgctcca gctcggaat cctccgact cctcttcaa cttgtggct aaaccaccg ggaaaattt aatcgaaaa catcgcaag cttgaagga tttctggcc cgtgatggc ggacccgtt tttctgcga ctataagca agacattga aatttacac aatcatcct ccgcataat taacttgga aatcggggc taacataga caaggtgga cgaaaaact cgtaaaaac catcgataa gtttttgaa ggacaagaa acgcattgc acagtggaa caaattcgt gcttgcgtc ctctcaaag cagtgcgaa aaacaagct ggacttgct cagtgcctg gatggagtt tttggtttt gacgaagaa agttatacc tgatagttt cgagaacga cgttaagtt cgccctcgt tgcctgttg tctggacgg cttgcaccc tcctgaagg cgggttcag taacatcga aacgattta tctgtggtc acagctgta cttgctgtt aataagtac gtggtcagt ctacgagaa aaaggagca agtggtttt ggggaagct acaaacgtt cctgcacgc gcttaacac gtattataa atatctgac tccgaaaat caaggagaa tattttatg tgcggtgaa tcggactat cctcgatct gcaggagta cttcagtgt taatacagc gcagttgtt gacattttt gtactccgt tgggccgct gattgacgc ggaatatta ctacctggt gtccgaagc gttcgagac tgaggacga tgcggaaaa agtgcggcg acttaagcc gtggttgat tattgtttt tatcggaaa ctcgataat ggcgctcga tcgctgcga ggaaattgg gttgtggtt ctactacga gcagttttt gcaacgggt gatggactc tgtggggtc aatgatgga acatagcgc cactttgcc gtttgcaaa gctagctat tgactttct tatcactta tgcggagtc tccgttcgt taaggactt cctaaagga ggatctgtt cagtttcta ctttaaagt gcgaccgcc accgctcac aataagtgt gggggaggc aattctatc gggacttcc ggtcaattt gaaggagtg gtggtttat tttaatcac tttgataaa gctgaaccg tattttgat acaacagat cggcgatgc aatgctaag gacgtgctt cacgttcat cacacaaca cgacttgtt gctcaagga aattgtttc tttgaccaa atacaccgt tgatatgtc ggccttgac tctcgtttg tgaaacttt gaaacttat tctggttgt tctgacaca aatgccaca ttggcgagt cgactcgcc gtactctta caacttaat aatggaggg tataaaaac gacaataca cgcctgcgt tttgtcaat cctggggta caaaaaaat taattttta tgacatcat caacggaaa aaatctgaa cattattga cgtcacccc aaccgactt actagtctc agtgttgaa taaactaac agaaattac gtactgggg ctgcagctg tttgaagaa aaccaagac cgacttagt aacccagct aaacgccca ctacgagac tgaaaacct cgcattatt attggtcga aaacgactt tagtgttcc ccgatttga gttgccgat cactccgag actgacgta cgggtctct attgtgcct ggctgacta tttgtgcaa aaccctcaa tcagttgtc tcctgcaac tccttcgag aatttcgag aagtcgtca agtgtctct aaccgatct taacaatca aatgaaagt caagagttt ggggaaaat tttcgagca ttgcaccgc aacgtatga ggaatttat aggttttat ggtctacat ccaaaaaag cgacgatct tccgaatga aaagttcag ttcgttttt gcactacga catgacgga attttccgg gacgacttc ggcggtttc ccagttgga ttatcattt ggttaagaa cgcgctaga ggcgttgat gagtttcct ggctgggga aatcttctt cgcaattcg gaatctccg aggcgaagc tgtatttgc atacaggag ggagttgag ttcggaaat acagttctt ctacgagtt tgtccgaaa gcggactag tgaacaata cagtgcgat ggtggcaac acctcgatc aacgtcctc catcgcaca gactgagcc cgaccttat caaaagata catcatcac caaagtcat ggataacga taccagcga tattgttga taataattt tgtacttgt tatttctca gtggttaat caaattctg ccacttgtc aaattaaat tgtatttcg tgtatcagg tttcaaata aaaaatatt ttataaaaa tacatttat ttcaccaca gactcaaca attttacgg aagtaacac aaaggcgta gctgtgttg agctccgcc catttcaac taattcgaa acatatttt cgttaccaa ttttgtcgc agc
