| General information | |||
| TC006201 | 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_007423.3: 4915173..4939130 | |
| 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 |
|
TC006201 | |
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_04085 | 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)
Analysis of larval stage 1 cuticle
number of eggs/embryos/larvae on slide: > 50
embryo/egg without visible larval cuticle in cuticle preparation - Penetrance (percentage of animals/eggs on slide): less than 30%
cuticle remnants without visible segmentation, differentiation or axis - Penetrance (percentage of animals/eggs on slide): less than 30%
head not present - Penetrance (percentage of animals/eggs on slide): 30% - 50%
thorax not present - Penetrance (percentage of animals/eggs on slide): 30% - 50%
abdomen not present - Penetrance (percentage of animals/eggs on slide): 30% - 50%
affected eggs/embryos/larvae total percentage of affected eggs/embryos/larvae on slide - Penetrance (percentage of animals/eggs on slide): 50% - 80%
embryo/egg without visible larval cuticle in cuticle preparation - Penetrance (percentage of animals/eggs on slide): less than 30%
cuticle remnants without visible segmentation, differentiation or axis - Penetrance (percentage of animals/eggs on slide): less than 30%
head not present - Penetrance (percentage of animals/eggs on slide): 30% - 50%
thorax not present - Penetrance (percentage of animals/eggs on slide): 30% - 50%
abdomen not present - Penetrance (percentage of animals/eggs on slide): 30% - 50%
affected eggs/embryos/larvae total percentage of affected eggs/embryos/larvae on slide - Penetrance (percentage of animals/eggs on slide): 50% - 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: 10.0% (includes death as larva, prepupa, pupa)
Lethalities 22 days after larval injection: 10.0% (includes death as larva, prepupa, pupa, adult)
- >TC006201 RA
MEDLLLNIV KESTGTKLS NLKQSAQEA HDLLCNQNN LLRSPSHEL RTACFIPLK LALESKKSK LVSLALTGL NKIVRDERF QSGTEPEDD SLWLPAQLL LATSSMLTQ CEDTQVHIL RVVLNLACS ASWALNGRL VMLLITRCG EAYENGTQP VRAAAQAAA SQTLTAFCT FLDEECQEI LQQQQKHKN SGSSAEMVY TKTSAVACF NEAIPVMQY LCSRLEEAK TQPKSADLT IFLLECLLT LVNTLPHTV HSNIHFTTF LWQRFCPAL LSLLGAPGD PTGHSLTTN QSKIIYSIG IQVVRLVGR ERALRPVLE ALFHRMLLL PSPLKRLEP LRAARELLR SPGRLADLL LLSGAIHRH AGDDMAIIR LIMDSIEES SQCSDTNVL LASVECVGA LLGDVVANL KCQSDSDSS GIEGNVPDI EGGSESDCS GATEGPEEY GSQSEDSIM DDDSFLTNH QLQKLHKLP KSLNLGRSG LEECNTDVE RHNARHFVK TLQNILLPN LLSLRSSIQ VDEVLQEFA SKCCQHNSA QSYEISTIM NADGVYLAT YSALLLDLK LIQGGNHDG GNNVPITEA QFVEEIHGS GVLVYVSAT WLCELYQNV LAKSLLLTA GYDPKAPQQ PALVNLLTD VGGLCPSQL LSDWQKLQK VHGSPESSP EVEAGVKLS RRVLTCCWA SVVSVLGAP LGEKSTFSG TSALSRLVA RRARQKHRQ RIRDDIITA CLEGLHKAA SLSNTLKLQ SRSCSILSL LSASSCKPQ GAKLPASHA LSLDVLLSK GLALGSHSA ACWPHVFSA CVAVANLEH ALFSKTGST QLLMVAQNT QNNTVTSST YTNSERLNM TFNISNDEE TCVDVYSFL HNASYAATN SSDASIAEI VERSGAHNA QSQGILRGV HAAKVCCVL SQKADELFH SAALRLSLP GLCSFLTEL CKASHTQLF TRYEDTPRQ QKKWWKKEL ESQQKPPAT LLLHRIGEV TLKCIRSGR PLIHTMKVW SIVGPHFMQ AACHKDRTI SKKAVQCIH DAVTALLNE QAELPHFHF NEALIKPFE NLLCLELCD LDVQDQIVS CLCEFVEAN RTEICSGWR PLFGTLRVT NAKNNAAAI LDVFRIFLS TDNTLVFAN AALDYILCL LSHIKNANF EENEPQVKQ QESKKGAFL DKDDFASKL VIGPVDLCI ESLKLLQNC AAILSMMYN MPKCPSFNV THRMNIDTD PQVVDPIIQ NIEVVNFNQ DHYDPISYS ILGTRHDLE LCESSGITL AKMDRQSNV LKIWYILLE GLSSASIMS AHKNQPFVV ETLFKLLRD LIDNPGVNF GLYSINHLL LPMVQNWMR QNSKIQKPG EIWQNFKHC CGLASELVV DYLHHLQGF GPFVENPAA TLALKQLLL ILIECITQP SENIARLGT SCIRHVILS AGKILTGGQ WEILVVALH RACTISLNP LHQLTLAFK ENSDSFYGD LATVKVAAR KDSTVSENE RLYELAQQV FLMQCQRNC LKCDKLCEC DASMTIDDR SYVFLLYPL DMSSILNPD LYTVRVPFR NLVVGILAH QMLVQTISS ALLQNLNHI TPILNILQI NSCSLRGIL THVNAKHVD ILLKCLDVS NARAKQFDV RPGLKFLTQ KVGNLSKAA NLYTQANTS EVVQIIVLI ELCLDGIEK YSIKPKDLK ELLGKDGKV RCSSDLDYV EQFLRRLQN KWEFLCESY INLTINIPV QDSEDELKS SSDNLDQIA DKPRPFKLA DFKQEEDTL SSSTDSEGY LEKEVLPNV KTEDEAEKP KLRQSCSLD ETILKKTKE VSSTDDETN DMAAKYTKQ IHDKYKVEL NSIKYDTNT LLQMRKSTI SSDNITSCD SFEKKTPSP RINPFMVAN VPPPQPIPP EIQQQRAMS IFKDSEAYK TTRIETMEA CLELLSSLS SEKLSPLAT VLKQGAVLL IGAQDEKIK NAAENLLNR MNISYIDHD NF
- >TC006201 RA
atggaagat ttactcctc aatattgtt aaagaatca acgggaacg aaattgtcg aatttaaaa cagagcgca caagaagcc cacgatctg ctatgcaac cagaacaac ctcctccgc agccccagc cacgagcta cgaaccgcc tgctttatt cctctgaaa ttggccctc gaaagcaaa aagagcaaa cttgtgtcg ctggccctg acaggactc aataaaatc gtgagggac gagcgcttc caatcgggc actgaacca gaagatgat tctctctgg ctccccgcg cagttactc ctcgctaca tcttctatg ctaacacaa tgcgaggac acacaagtc cacatcttg cgcgtagtc ctaaacctg gcatgttca gcaagctgg gccttgaac ggccgtttg gtcatgctt ctgataaca cgatgtgga gaggcctac gaaaacggt acacaacct gtccgagca gctgctcaa gccgcagcg agccaaact ctgaccgcc ttttgcacc ttcttggac gaagagtgc caagaaatc ttacaacag cagcaaaag cacaaaaat tcaggcagc agtgccgag atggtgtac acgaaaacg tcggcagtt gcgtgtttt aacgaagca attcctgtt atgcaatac ttgtgcagc agactagaa gaggccaaa acgcaacca aaatcggcc gatcttacg atttttctc cttgagtgt ctcctaacc cttgtcaac acccttcca cacaccgtt cattctaac atccacttc accactttc ctttggcaa cgcttttgt cccgccctt ttgtccctt ctgggtgct ccgggtgac ccaactggg cattccctc accaccaat cagagcaaa atcatctac agtatcggt atccaagtc gtgcgtctt gttggtcgt gaacgtgcc cttcgtcct gtccttgaa gcccttttc catcgtatg ctccttctc ccgagtcca ctcaaaaga cttgaacct ttaagggcc gcccgtgaa ctgttaagg tcgccaggg cgtctcgct gatcttttg ctacttagt ggggctatt catagacat gcgggggac gatatggct ataatccga ctaattatg gattcgatt gaggagtcg tcacaatgt agcgacacg aatgtactt ttggcgagt gttgagtgt gtgggggcg cttttaggg gacgtggtt gcaaatttg aaatgccaa agcgactcc gacagtagc gggattgag ggcaacgtc cccgacatt gaagggggc tctgaatcg gattgttcc ggcgcaacc gaaggcccc gaagaatac gggagccaa tcagaggac agtataatg gacgatgac agtttttta accaaccat caactccaa aaattgcac aaacttcca aaatcgctc aacttgggc cgaagtggt ctagaagag tgcaatacc gacgtggaa cgtcacaac gcccgccat ttcgtcaaa accttacaa aacatactt ctcccaaac cttttatca ctaagatcg tccattcaa gtcgacgaa gttttgcaa gaattcgcc tccaagtgt tgccaacac aactcggcc caaagctac gaaatttca acgattatg aacgccgat ggtgtttat ttagccacg tattcggcc cttttgctc gacttgaag ctgatacaa gggggcaac catgatggg ggcaacaac gtccccatc acggaggcc cagtttgtg gaggagatt catggcagt ggggttttg gtctacgtt tcggcgact tggttatgc gaactttac caaaatgtt ttggccaaa tcgttgctt ctgacggct gggtatgac cccaaagcg ccccaacag ccagctttg gtcaattta ttgacagat gttggtggt ttatgtccg tcacagtta ctttccgat tggcagaag ttgcaaaaa gtccatgga agcccggag agtagtcct gaagttgag gcaggagtt aagctctca cggagggtt ttgacgtgt tgttgggct tccgtggtt tcggtttta ggggcgcca ttgggcgag aaaagcact ttttcgggg acttcggct ctgagtcgg ttggtggcg agaagggcg cggcagaag catcggcaa aggataagg gatgatatc attacggcg tgtcttgag ggcctccat aaggcagca agcttgagt aacactttg aaactccaa tcccggagc tgcagtatt ttatcgcta ctttcggct tcttcctgt aagccacaa ggcgccaaa ctccctgct tcccacgcc ctttctttg gacgtcctt ttatccaaa gggctagcc cttggctcg cacagtgcc gcctgttgg ccacatgtc ttctccgct tgcgtggcc gtagccaac ctcgaacat gcccttttt agcaaaact ggctcaact caactcttg atggttgcc caaaacacg caaaacaat accgtgaca tcgagcact tacaccaac agtgagaga cttaacatg actttcaat atcagtaac gacgaagag acttgcgtg gacgtttac agttttctg cataatgcc tcgtatgcc gccacgaat tcgtctgat gcctcgatt gccgaaatt gtggaacgc agtggggca cataatgcc caaagtcag ggcattttg agaggagtt catgcagcg aaagtttgc tgcgttttg tcgcagaaa gcagatgag ttgtttcat tcagctgcg ttgaggttg tctctgcca gggctttgc agtttcttg acggagttg tgcaaggcg tcgcataca cagctcttt acccgttat gaggacacc cctcgccag cagaaaaaa tggtggaaa aaagagctc gaatcccag caaaaaccc ccagccact ctcctcctc caccgaatc ggtgaagta accctcaag tgcatccgt tccggccgc cctttaatc cacaccatg aaagtctgg tcgatcgtc ggcccccat ttcatgcaa gcagcgtgt cacaaagac cgcacaatc tccaaaaaa gcagtccaa tgcatccat gacgcagtc accgcccta ttaaacgaa caagccgaa ctgccacac ttccacttc aacgaagcc ctaatcaaa ccattcgaa aacctcctc tgtctggaa ctgtgcgac ttggacgtc caagaccaa atcgtgtcc tgcctttgc gagttcgtg gaagccaac aggacagag atctgctcg ggttggagg ccacttttc gggacttta cgagttacc aatgcgaaa aataacgca gctgcgatt ttggacgtg ttccggatt ttcctctcg actgataat acgctggtg tttgcgaac gcggctttg gactacatt ttgtgtctt ttgtcacac atcaaaaac gccaatttt gaggaaaac gagccgcaa gttaaacaa caagagagt aagaaaggg gcgtttttg gataaagat gattttgcc tcaaaactg gtcatcgga ccggttgat ctatgcatc gaaagcctc aaactcctc caaaattgc gccgccatc ttatcaatg atgtacaac atgccgaaa tgtccctcc tttaacgtc acacatcgc atgaacatt gacaccgat ccgcaagtt gttgatcca atcatccag aatattgaa gttgttaac ttcaaccaa gaccattac gacccgatt agttactcg attttgggg accagacac gacttagaa ttgtgcgaa tcaagtggt attacttta gccaaaatg gatcggcaa agtaacgtt ttgaaaatc tggtacatt ctccttgaa ggactctcc tcagcttct atcatgtca gctcataaa aaccaacca tttgttgtt gaaaccctc ttcaaactt ttaagggat ttgattgat aacccgggc gttaacttt gggttatat tccataaat cacttgcta ctcccgatg gttcaaaac tggatgcgc cagaacagc aagatccaa aaaccgggc gaaatctgg caaaacttc aagcactgt tgtggctta gctagcgag cttgtggtc gattatttg caccatttg caaggtttt ggccctttt gttgaaaac ccagccgct actttggcc ctcaaacaa ctcctcttg atcctaatc gagtgtata acacaacct agcgaaaac atagcccgg ttagggact tcctgcatc cgtcacgtt attttaagt gctggtaag attttgact ggagggcaa tgggaaatt ctggtcgtg gccctacac agggcttgc accatctcg cttaaccct ctccaccaa ctaactctc gcatttaaa gagaactcg gatagtttt tacggcgac ttggcgaca gttaaagtc gcagcgcgt aaagacagc acagtgtcc gaaaacgag aggttgtac gagcttgcc cagcaagtt ttcctcatg caatgccaa cgcaactgt ctcaagtgc gacaagttg tgcgagtgt gatgccagt atgacgatt gatgatcgc agttatgtt ttcctcctc tatccgctt gacatgtct tcgattttg aaccccgat ttgtataca gtgcgggtt ccatttcgc aatttggtt gttgggatt ttggcgcat caaatgctc gttcagacg atttccagc gcgcttttg caaaatttg aatcatatc acgccgatt ttgaacatt ttgcaaatc aattcgtgc agtttaagg gggattttg acgcatgtt aacgccaaa catgtcgat attctgttg aagtgtttg gatgtgagt aatgccaga gcgaagcag tttgatgtc aggcccggt ttgaaattt ttgactcag aaggttggg aatttgagc aaggcggcg aatttgtac acacaggca aacacttca gaggtggtc caaatcatt gttttaatc gagttgtgt cttgatggc attgagaaa tattcaatc aaacccaaa gacttgaaa gaattgttg gggaaggat ggtaaagtt aggtgttct agtgatttg gactacgtg gagcaattt ttgcgacgg ttgcaaaac aagtgggag tttttgtgc gaatcttac atcaaccta acgataaac atcccagtg caagacagt gaagacgaa ttgaagagt tctagtgat aatcttgac caaattgcc gacaaaccc cgacctttc aaattagca gatttcaag caagaagaa gacactttg agttcaagc actgattcc gaaggttac ttggagaag gaagttttg ccaaatgtg aaaactgaa gatgaggca gaaaaaccc aaactacgt caaagttgt agtcttgac gagacaatt ttgaaaaag acaaaggaa gtctcatcg actgatgac gaaacgaat gacatggcg gccaaatat acgaaacag atccacgac aagtataaa gttgagctt aattcgatt aaatacgac actaatacg ttgcttcaa atgcgtaaa tcgaccatt tcgagtgat aatattacg agttgtgac agttttgag aaaaagacg ccaagtccg aggattaat ccttttatg gtggctaat gtgccaccg ccacagccg ataccgccc gaaatacag caacagagg gccatgagt atctttaag gactctgag gcgtacaag acgactcgg atcgagaca atggaggca tgtctcgag cttttgagt agtctatcg tctgaaaag ctctcgcct ttggccacc gttttgaaa caaggcgcc gttcttcta atcggggct caggacgag aaaataaaa aacgcagct gagaatctt ctcaacaga atgaatatc tcgtacatt gaccacgat aatttctaa
- >TC006201 RA
tcaggcaag tattgattt tgtccagac atgctcagt cacaacaac cagtcatca atccgcaat atgttagcc gcttaatca ccacacaat ggaagattt actcctcaa tattgttaa agaatcaac gggaacgaa attgtcgaa tttaaaaca gagcgcaca agaagccca cgatctgct atgcaacca gaacaacct cctccgcag ccccagcca cgagctacg aaccgcctg ctttattcc tctgaaatt ggccctcga aagcaaaaa gagcaaact tgtgtcgct ggccctgac aggactcaa taaaatcgt gagggacga gcgcttcca atcgggcac tgaaccaga agatgattc tctctggct ccccgcgca gttactcct cgctacatc ttctatgct aacacaatg cgaggacac acaagtcca catcttgcg cgtagtcct aaacctggc atgttcagc aagctgggc cttgaacgg ccgtttggt catgcttct gataacacg atgtggaga ggcctacga aaacggtac acaacctgt ccgagcagc tgctcaagc cgcagcgag ccaaactct gaccgcctt ttgcacctt cttggacga agagtgcca agaaatctt acaacagca gcaaaagca caaaaattc aggcagcag tgccgagat ggtgtacac gaaaacgtc ggcagttgc gtgttttaa cgaagcaat tcctgttat gcaatactt gtgcagcag actagaaga ggccaaaac gcaaccaaa atcggccga tcttacgat ttttctcct tgagtgtct cctaaccct tgtcaacac ccttccaca caccgttca ttctaacat ccacttcac cactttcct ttggcaacg cttttgtcc cgccctttt gtcccttct gggtgctcc gggtgaccc aactgggca ttccctcac caccaatca gagcaaaat catctacag tatcggtat ccaagtcgt gcgtcttgt tggtcgtga acgtgccct tcgtcctgt ccttgaagc ccttttcca tcgtatgct ccttctccc gagtccact caaaagact tgaaccttt aagggccgc ccgtgaact gttaaggtc gccagggcg tctcgctga tcttttgct acttagtgg ggctattca tagacatgc gggggacga tatggctat aatccgact aattatgga ttcgattga ggagtcgtc acaatgtag cgacacgaa tgtactttt ggcgagtgt tgagtgtgt gggggcgct tttagggga cgtggttgc aaatttgaa atgccaaag cgactccga cagtagcgg gattgaggg caacgtccc cgacattga agggggctc tgaatcgga ttgttccgg cgcaaccga aggccccga agaatacgg gagccaatc agaggacag tataatgga cgatgacag ttttttaac caaccatca actccaaaa attgcacaa acttccaaa atcgctcaa cttgggccg aagtggtct agaagagtg caataccga cgtggaacg tcacaacgc ccgccattt cgtcaaaac cttacaaaa catacttct cccaaacct tttatcact aagatcgtc cattcaagt cgacgaagt tttgcaaga attcgcctc caagtgttg ccaacacaa ctcggccca aagctacga aatttcaac gattatgaa cgccgatgg tgtttattt agccacgta ttcggccct tttgctcga cttgaagct gatacaagg gggcaacca tgatggggg caacaacgt ccccatcac ggaggccca gtttgtgga ggagattca tggcagtgg ggttttggt ctacgtttc ggcgacttg gttatgcga actttacca aaatgtttt ggccaaatc gttgcttct gacggctgg gtatgaccc caaagcgcc ccaacagcc agctttggt caatttatt gacagatgt tggtggttt atgtccgtc acagttact ttccgattg gcagaagtt gcaaaaagt ccatggaag cccggagag tagtcctga agttgaggc aggagttaa gctctcacg gagggtttt gacgtgttg ttgggcttc cgtggtttc ggttttagg ggcgccatt gggcgagaa aagcacttt ttcggggac ttcggctct gagtcggtt ggtggcgag aagggcgcg gcagaagca tcggcaaag gataaggga tgatatcat tacggcgtg tcttgaggg cctccataa ggcagcaag cttgagtaa cactttgaa actccaatc ccggagctg cagtatttt atcgctact ttcggcttc ttcctgtaa gccacaagg cgccaaact ccctgcttc ccacgccct ttctttgga cgtcctttt atccaaagg gctagccct tggctcgca cagtgccgc ctgttggcc acatgtctt ctccgcttg cgtggccgt agccaacct cgaacatgc cctttttag caaaactgg ctcaactca actcttgat ggttgccca aaacacgca aaacaatac cgtgacatc gagcactta caccaacag tgagagact taacatgac tttcaatat cagtaacga cgaagagac ttgcgtgga cgtttacag ttttctgca taatgcctc gtatgccgc cacgaattc gtctgatgc ctcgattgc cgaaattgt ggaacgcag tggggcaca taatgccca aagtcaggg cattttgag aggagttca tgcagcgaa agtttgctg cgttttgtc gcagaaagc agatgagtt gtttcattc agctgcgtt gaggttgtc tctgccagg gctttgcag tttcttgac ggagttgtg caaggcgtc gcatacaca gctctttac ccgttatga ggacacccc tcgccagca gaaaaaatg gtggaaaaa agagctcga atcccagca aaaaccccc agccactct cctcctcca ccgaatcgg tgaagtaac cctcaagtg catccgttc cggccgccc tttaatcca caccatgaa agtctggtc gatcgtcgg cccccattt catgcaagc agcgtgtca caaagaccg cacaatctc caaaaaagc agtccaatg catccatga cgcagtcac cgccctatt aaacgaaca agccgaact gccacactt ccacttcaa cgaagccct aatcaaacc attcgaaaa cctcctctg tctggaact gtgcgactt ggacgtcca agaccaaat cgtgtcctg cctttgcga gttcgtgga agccaacag gacagagat ctgctcggg ttggaggcc acttttcgg gactttacg agttaccaa tgcgaaaaa taacgcagc tgcgatttt ggacgtgtt ccggatttt cctctcgac tgataatac gctggtgtt tgcgaacgc ggctttgga ctacatttt gtgtctttt gtcacacat caaaaacgc caattttga ggaaaacga gccgcaagt taaacaaca agagagtaa gaaaggggc gtttttgga taaagatga ttttgcctc aaaactggt catcggacc ggttgatct atgcatcga aagcctcaa actcctcca aaattgcgc cgccatctt atcaatgat gtacaacat gccgaaatg tccctcctt taacgtcac acatcgcat gaacattga caccgatcc gcaagttgt tgatccaat catccagaa tattgaagt tgttaactt caaccaaga ccattacga cccgattag ttactcgat tttggggac cagacacga cttagaatt gtgcgaatc aagtggtat tactttagc caaaatgga tcggcaaag taacgtttt gaaaatctg gtacattct ccttgaagg actctcctc agcttctat catgtcagc tcataaaaa ccaaccatt tgttgttga aaccctctt caaactttt aagggattt gattgataa cccgggcgt taactttgg gttatattc cataaatca cttgctact cccgatggt tcaaaactg gatgcgcca gaacagcaa gatccaaaa accgggcga aatctggca aaacttcaa gcactgttg tggcttagc tagcgagct tgtggtcga ttatttgca ccatttgca aggttttgg cccttttgt tgaaaaccc agccgctac tttggccct caaacaact cctcttgat cctaatcga gtgtataac acaacctag cgaaaacat agcccggtt agggacttc ctgcatccg tcacgttat tttaagtgc tggtaagat tttgactgg agggcaatg ggaaattct ggtcgtggc cctacacag ggcttgcac catctcgct taaccctct ccaccaact aactctcgc atttaaaga gaactcgga tagttttta cggcgactt ggcgacagt taaagtcgc agcgcgtaa agacagcac agtgtccga aaacgagag gttgtacga gcttgccca gcaagtttt cctcatgca atgccaacg caactgtct caagtgcga caagttgtg cgagtgtga tgccagtat gacgattga tgatcgcag ttatgtttt cctcctcta tccgcttga catgtcttc gattttgaa ccccgattt gtatacagt gcgggttcc atttcgcaa tttggttgt tgggatttt ggcgcatca aatgctcgt tcagacgat ttccagcgc gcttttgca aaatttgaa tcatatcac gccgatttt gaacatttt gcaaatcaa ttcgtgcag tttaagggg gattttgac gcatgttaa cgccaaaca tgtcgatat tctgttgaa gtgtttgga tgtgagtaa tgccagagc gaagcagtt tgatgtcag gcccggttt gaaattttt gactcagaa ggttgggaa tttgagcaa ggcggcgaa tttgtacac acaggcaaa cacttcaga ggtggtcca aatcattgt tttaatcga gttgtgtct tgatggcat tgagaaata ttcaatcaa acccaaaga cttgaaaga attgttggg gaaggatgg taaagttag gtgttctag tgatttgga ctacgtgga gcaattttt gcgacggtt gcaaaacaa gtgggagtt tttgtgcga atcttacat caacctaac gataaacat cccagtgca agacagtga agacgaatt gaagagttc tagtgataa tcttgacca aattgccga caaaccccg acctttcaa attagcaga tttcaagca agaagaaga cactttgag ttcaagcac tgattccga aggttactt ggagaagga agttttgcc aaatgtgaa aactgaaga tgaggcaga aaaacccaa actacgtca aagttgtag tcttgacga gacaatttt gaaaaagac aaaggaagt ctcatcgac tgatgacga aacgaatga catggcggc caaatatac gaaacagat ccacgacaa gtataaagt tgagcttaa ttcgattaa atacgacac taatacgtt gcttcaaat gcgtaaatc gaccatttc gagtgataa tattacgag ttgtgacag ttttgagaa aaagacgcc aagtccgag gattaatcc ttttatggt ggctaatgt gccaccgcc acagccgat accgcccga aatacagca acagagggc catgagtat ctttaagga ctctgaggc gtacaagac gactcggat cgagacaat ggaggcatg tctcgagct tttgagtag tctatcgtc tgaaaagct ctcgccttt ggccaccgt tttgaaaca aggcgccgt tcttctaat cggggctca ggacgagaa aataaaaaa cgcagctga gaatcttct caacagaat gaatatctc gtacattga ccacgataa tttctaatt tatttgcac tctttaaga ctggatttg gaagtctga tggtaaatc ttcgtgctt actttccgt ataattact gtagaataa aaaataaat gtaaaa
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