Difference between revisions of "Os02g0709400"
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===Function=== | ===Function=== | ||
Please input function information here. | Please input function information here. | ||
| + | The rice (Oryza sativa L.) basic leucine Zipper factor RISBZ1 and rice prolamin box binding factor (RPBF) are transcriptional activators of rice seed storage protein (SSP) genes in vivo. Storage lipids were accumulated at reduced levels in KD-RISBZ1 and KD-RISBZ1/KD-RPBF seeds. KD-RPBF and KD-RISBZ1/KD-RPBF seeds exhibited multi-layered aleurone cells. Gene expression of DEFECTIVE KERNEL1 (OsDEK1), CRINKLY4 (OsCR4) and SUPERNUMERARY ALEURONE LAYER 1 (OsSAL1) rice homologues was decreased in the KD mutants, suggesting that these genes are regulated by RISBZ1 and RPBF. These phenotypes suggest that combinatorial interactions between RISBZ1 and RPBF play an essential role during grain filling. The functional redundancy and compensation between RISBZ1 and RPBF possibly account for weak effects on the SSP levels in single KD mutants, and help maintain various processes during seed development in rice. Physical interaction between RISBZ1 and RPBF may ensure that these processes are carried out properly. | ||
| + | DEK1 and is a calpain-like proteinase and a receptor-like kinase,(Becraft et al., 1996; Lid et al., 2002). Both DEK1 binds to the plasma membrane, and is involved in the production of the aleurone layer specification signal. The multiple aleurone layer phenotype in sal1 is thought to be caused by an overabundance of aleurone membrane signaling proteins, such as DEK1 and CR4 (Tian et al., 2007).Previous studies on maize and Arabidopsis,CRINKLY 4 (CR4), DEFECTIVE KERNEL 1 (DEK1) and SUPERNUMERARY ALEURONE LAYER 1 (SAL1) have shown them to be key players in determining the number of aleurone layers (Becraft et al., 1996; Lid et al., 2002; Shen et al., 2003).The expression of both positive (OsCR4 and OsDEK1) and negative (OsSAL1) determining genes for the aleurone layer number was decreased in KD-RISBZ1 and KD-RPBF, and significantly decreased in KD-RISBZ1/KD-RPBF seeds, suggesting that both RISBZ1 and RPBF are positive regulators of these genes. | ||
===Expression=== | ===Expression=== | ||
Revision as of 12:22, 9 June 2014
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Contents
Annotated Information
Function
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The rice (Oryza sativa L.) basic leucine Zipper factor RISBZ1 and rice prolamin box binding factor (RPBF) are transcriptional activators of rice seed storage protein (SSP) genes in vivo. Storage lipids were accumulated at reduced levels in KD-RISBZ1 and KD-RISBZ1/KD-RPBF seeds. KD-RPBF and KD-RISBZ1/KD-RPBF seeds exhibited multi-layered aleurone cells. Gene expression of DEFECTIVE KERNEL1 (OsDEK1), CRINKLY4 (OsCR4) and SUPERNUMERARY ALEURONE LAYER 1 (OsSAL1) rice homologues was decreased in the KD mutants, suggesting that these genes are regulated by RISBZ1 and RPBF. These phenotypes suggest that combinatorial interactions between RISBZ1 and RPBF play an essential role during grain filling. The functional redundancy and compensation between RISBZ1 and RPBF possibly account for weak effects on the SSP levels in single KD mutants, and help maintain various processes during seed development in rice. Physical interaction between RISBZ1 and RPBF may ensure that these processes are carried out properly. DEK1 and is a calpain-like proteinase and a receptor-like kinase,(Becraft et al., 1996; Lid et al., 2002). Both DEK1 binds to the plasma membrane, and is involved in the production of the aleurone layer specification signal. The multiple aleurone layer phenotype in sal1 is thought to be caused by an overabundance of aleurone membrane signaling proteins, such as DEK1 and CR4 (Tian et al., 2007).Previous studies on maize and Arabidopsis,CRINKLY 4 (CR4), DEFECTIVE KERNEL 1 (DEK1) and SUPERNUMERARY ALEURONE LAYER 1 (SAL1) have shown them to be key players in determining the number of aleurone layers (Becraft et al., 1996; Lid et al., 2002; Shen et al., 2003).The expression of both positive (OsCR4 and OsDEK1) and negative (OsSAL1) determining genes for the aleurone layer number was decreased in KD-RISBZ1 and KD-RPBF, and significantly decreased in KD-RISBZ1/KD-RPBF seeds, suggesting that both RISBZ1 and RPBF are positive regulators of these genes.
Expression
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Evolution
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Labs working on this gene
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References
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Structured Information
| Gene Name |
Os02g0709400 |
|---|---|
| Description |
Dek1-calpain-like protein |
| Version |
NM_001054425.1 GI:115448220 GeneID:4330484 |
| Length |
14893 bp |
| Definition |
Oryza sativa Japonica Group Os02g0709400, complete gene. |
| Source |
Oryza sativa Japonica Group ORGANISM Oryza sativa Japonica Group
Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
Spermatophyta; Magnoliophyta; Liliopsida; Poales; Poaceae; BEP
clade; Ehrhartoideae; Oryzeae; Oryza.
|
| Chromosome | |
| Location |
Chromosome 2:30229855..30244747 |
| Sequence Coding Region |
30229910..30230037,30230459..30230709,30231188..30231370,30231467..30231602,30231745..30231875 |
| Expression | |
| Genome Context |
<gbrowseImage1> name=NC_008395:30229855..30244747 source=RiceChromosome02 preset=GeneLocation </gbrowseImage1> |
| Gene Structure |
<gbrowseImage2> name=NC_008395:30229855..30244747 source=RiceChromosome02 preset=GeneLocation </gbrowseImage2> |
| Coding Sequence |
<cdnaseq>atggaagaggaggaacaccgcggggttgttcttgtttgcagcatctgcggtttcctcttcgctgtccttggccctctcagcttttggatcctatgggctgtgaactggaggccgtggaggctatatagttggatatatgcaaggaaatggccagcatatgtgcaaggacctcaactaagcacactgtgcagctttttcactctttttgcatggcttgtcgtcgtttctccaataacagttttgttagtatggggaggcatccttattgctcttttggaaaggaacataattgggctagctgttataatggtcggtgttgctttgcttctgtcattctactctataatgttgtggtggagaacgcaatggcaaagctccaaggctgttgcttatcttctactcctggcagtcggtctactgtgcgcttatgagttttgtgctgtttatgttacaaccggtgctagtgcttctgagctcaattccccatctggcttcttctttggggtatctgcaatttcattggccataaatatgctttttataagtaaaatactgttcaatggaagtggattcgatgttgatgagtatgtccgaaggttgtacaaatttgcatattctgattgtgttgaagtggcccctgtttcatgctcgcctgacccaccagatcctagtgagttatacatgacaaaatccagcagggtcttgcatctagggcttctttacctttgctcattgatggttcttgttgtctattccatcttatatggtctgacatcaaaagaagcacgctggctgggtgctttaacttcagttgcagtagtgattctcgactggaatttgggcctgtgttcatttaggtttgagcttcttaaaagtcggatgatagcgctatttgtagctggaacatcaagggttttccttatatgctttggggtgcactactggtaccttggacattgcatcagttatgcttttgtagcatctgtgcttttagctgcagctgtctcttgctggctttctatttcaaacccctcagttgcaaggatagatgcattaagaagcacagtgataaaactccgagaaggttttcgaaggaaggggcaaactagttcttcaaactcatcagatggctgtggatctagtgtgaagcgtagtagtggcagtgttgaagctggtcctcatggcaatgctactgattctatgtacagaagcaactcacaaagcgattgtgtcaattggaacaatgttccttttgatcgatcgaatagttgtcaagaaggacagagctctgacaagaacatagatagtggacgtgccagtttagcccatcgcagtaattcgtgcctttctgctgtagctgtacaagaccctgaaactgctgtagtttcagcggacaggcatggggacccaactgcttcactcgttgtttgctctagtagtggtttggaaagtcaaggctgcgagtctagtggatcagctactgcctcgggtaatcagcagctattggatttgaacttggctgcaatttttcaggacagactgaatgatccaagaatcacatctatgctaaaaaggaatggtggactaggagatgtagaattggctaatcttcttcaggataaaggccttgatccaaatttttcctatatgatgaaagacaaggttatggacccacgaattttagctttgctgcagagaagcagcttagatgcagacagagagcatcaagatgatgtagatgtcactggtactgattcagataggttagatacaactattgcaaaccagatctcgctatcagaggaacttaggagaagtgggctagaaaattggctaaatctttcaaggctgatgtttcatcaagtagcaggttcaccaatacgtgcttttgttgtttttaccctaatattcatcatagaaacagttactgtggctgtccatcgaccgaagcccatcaaggttataaatgcaactcatgaacagtttgagtttggtttctcgatactgcttctatcaccagttgtctgctccattatggcattcatttggtccctgtgtgcagaagagatgacgatgacatccaaacctcggaagtatggcttcatcgcatggttgctgagtacttgtgttggtctacttctgtctttcttgagcaagtcatcagttattttgggtttgtcactcacggtcccacttatggtggcttgtctctcatttgctattcccatatggatgcgtaacggttaccgtttctggattccgggaggggagcttgatagccgtgaaaatatccgccaagctccaggaaagaaagagcgcgctctgtttgccatcagcataactgttttcactgcatcagtaattggccttggtgcaatagtatcagcaaaacctctagatgcattgggctacaagggatgggatgctgataaaaagagtttctattctccctatgcaacatcgatgtatcttggatgggcattatcatcaacgattgctgtacttgccactggggtgatacctattgtcgcttggtttgctacttatcgattttcaccttcatcagctatatgtgttggcctttttgcaactgtccttgtatccttttgtggtgtgtcctactggggagtggtaaattcacgacaggatggagttcctctgaaggctgatttcctggcagctttacttccattactttgcattcctgctgtgttctcgttgtttactgggatgtataaatggaaggatgatgattggaagatttcccgtggagtttacctatttgttggcatgggagtgctgttattgcttggtgcaatatctgcagtaattgttacaattagaccttggactgttggagttgcatgcctccttgtcatcttgttccttgtatttgctattggtgtcattcactactggacatctaacaatttctatttaacaaggacacaaatgttacttgtttgctcccttgcttttctccttgccttggctgctttcctgatgggcttatttcaggagaaacctttcgttggggcatctataggttatttctcgtttctatttcttctcacaggaagagcattgactgtccttctatcaccaccaattgtcgtgtattcaccgagggtattacctgtttatgtttatgatgcccatgcagactctgctaaaaatgttagttatgccttccttatcttgtatgggattgcattggctactgaagtttggggtgtcattgccagtctaatattgaatccaccatttattggggctgctatttctgctatcactcttgtaattgccttcagttttgctgtttctcgaccatgcctgactctcaagatgctggaggacgctgttcactttctcagtaaggatacagttgtacaagcaatgtcacgttctgctaacaaaacgagaaatgctatttctgggacttactcggcgcctcagagatccgcaagttctgctgctcttttggttggagatcctgctattacgttggacagggctgggaattttgtgcttcctagggctgatgtcatgaagctgagagatcgtttaagaaatgaagaaattacggcaggatcgttcttttgtggagtgaagaattgtttaatgattggatcccctgtagatgtagattatcggaggaatatgtgtgcccatgcacgtattttggctttggaagaagcaattgatactgaatgggtttacatgtgggacaagtttggtggttatttacttcttctgcttggcttgactgccaaagctgagcaaatacaggatgaagttcgtcttagactttttctggacagcataggtctttctgacttaagtgccaaagaaatcaagaaatggatgcctgaagatcgaagacattttgaacttattcaagaaagctacataagggaaaaagaaatggaagaggaggtcttgatgcaaaggcgcgaggaagaagggaagggaagggaaaggaggaaggcattgttggagagagaggaacgaaaatggaaggagcttgaaatatcactgctatcttctattcctaatgctggaagcagggatgctgcagccatggcagcagctgtcagagctgtgggtggtgattctgctctggaagattcttttgcaagggacagggtctcgtcaatagctcgtcacataagaaaggcacaattggctcggcgagcagagcagactggcattccagacactgtttgcatacttgatgatgaaccaaggagtactgggcgtcattgtggagagattgatttgtgtctttgtgaaagtaaaaaagttagcttctccattgcagtaatggtgcagccggtgtcaggtccagtctgtctttttggaacagaattccagaaaaaagtgtgctgggaaatcttggtggcagggtcagaacagggaatggaagctggccaagttggtcttcgattggtgactaagggtgaaaggatgacaaccgttgcaaaagagtggaatattggtgcatcaagtattgctgatggcaggtggcatcttgtcactgtgactatagatgctgatctaggtgaagcaacttctttcatagacggagtctacgatggatatcagaatgcgttgccattgcctagaaataatggcatctgggaacctgggacagatatttgggttggtgctaggccacccacggacttagatgcctttggtaggtcagacagcgagggatcagactcaaagatgcagatcatggatgcttttctatggggaagatgtctgactgaagatgaggttgctatgttgcatacagccatctgttctgctgaatatggcctttttgatcttgctgctgaggatgcttggcatggaagttattctgcaagggtggatgactgggaaagtgaagaagcaaattttgaactatatgatcaagaggacgttgaatgggatggacagtactccagtggtaggaaacgtcatgcgcgtgattctgtagctattgacattgactcctttgctagaagaccaaggaaaccaagatttgagacacgagaggaagtcaatcagcgcatgctttctgtcgaaagggctgttagagaggctcttattgcgaagggagagagaaattttactgatcaagagttccctccggatgatcgatctctgtttgtagatccgatgaacccatctctaaaattgcaggttgtctctgagtggatgaggccttctgatattgcgaaggaggtatctatcagttcacaaccgtgcttattttctggttctgtgaattcgtcagatgtgtgtcagggtcggttgggcgactgttggttcttgagcgctgttgcagttttgacagagatggcccgtatatctgaagttataatcactcctgagtataatgaagaagggatttacactgtcagattctgtatccagggtgaatgggtggcagtggtggttgatgattggatcccctgtgagtctccggggaaaccggcatttgctactagtagaaagcaaaatgagctgtgggtatccattcttgaaaaagcttatgcaaagcttcatggctcttatgaggcattggagggtgggcttgtgcaagatgctctagtggatctcacaggaggagctggtgaagagattgacatgcggagcccccaagcgcagattgatcttgctagtggaagattgtggtctcagttgttgcattttaaacaagagggctttcttcttggtgctggaagtccttctggctcggatgctcatatttcatcaagtggcattgttcagggacatgcttactcaattttgcaggtaagagaagttgatggccataagctcgtgcaaattagaaatccgtgggcaaatgaagttgaatggaatggtccgtggtcagactcatcacaagagtggactgagcgaatgaagcacaaacttaagcatgttccacagtcaaagaatggggtattctggatgtcttggcaagattttcagatccactttcggtcaatatatgtctgtcgtgtttatccacccgagatgcgttactctgtccatggccaatggcgtggttatagtgcaggtggttgccaagattatgactcatggcatcaaaatcctcagtaccggcttagagtaacaggacgggatgcactgtatcctgtacatgtatttattacccttacgcagggtgttggtttttctagaaagacaaatggtttccggaactatcaatctagccatgactcttctatgttttacattggaatgaggatactcaagacacggggctgccgtgctgcttacaatatctacatgcatgaatcagtgggtggaacagattatgttaactcaagggagatatcatgtgaattagtgttggagccttatccaaaaggatacacaattgtgcccacaaccatccatcctggagaggaagcaccttttgttttatcagtttttacaaaagcaccaatcaagctagaagctgtttaa</cdnaseq> |
| Protein Sequence |
<aaseq>MEEEEHRGVVLVCSICGFLFAVLGPLSFWILWAVNWRPWRLYSW IYARKWPAYVQGPQLSTLCSFFTLFAWLVVVSPITVLLVWGGILIALLERNIIGLAVI MVGVALLLSFYSIMLWWRTQWQSSKAVAYLLLLAVGLLCAYEFCAVYVTTGASASELN SPSGFFFGVSAISLAINMLFISKILFNGSGFDVDEYVRRLYKFAYSDCVEVAPVSCSP DPPDPSELYMTKSSRVLHLGLLYLCSLMVLVVYSILYGLTSKEARWLGALTSVAVVIL DWNLGLCSFRFELLKSRMIALFVAGTSRVFLICFGVHYWYLGHCISYAFVASVLLAAA VSCWLSISNPSVARIDALRSTVIKLREGFRRKGQTSSSNSSDGCGSSVKRSSGSVEAG PHGNATDSMYRSNSQSDCVNWNNVPFDRSNSCQEGQSSDKNIDSGRASLAHRSNSCLS AVAVQDPETAVVSADRHGDPTASLVVCSSSGLESQGCESSGSATASGNQQLLDLNLAA IFQDRLNDPRITSMLKRNGGLGDVELANLLQDKGLDPNFSYMMKDKVMDPRILALLQR SSLDADREHQDDVDVTGTDSDRLDTTIANQISLSEELRRSGLENWLNLSRLMFHQVAG SPIRAFVVFTLIFIIETVTVAVHRPKPIKVINATHEQFEFGFSILLLSPVVCSIMAFI WSLCAEEMTMTSKPRKYGFIAWLLSTCVGLLLSFLSKSSVILGLSLTVPLMVACLSFA IPIWMRNGYRFWIPGGELDSRENIRQAPGKKERALFAISITVFTASVIGLGAIVSAKP LDALGYKGWDADKKSFYSPYATSMYLGWALSSTIAVLATGVIPIVAWFATYRFSPSSA ICVGLFATVLVSFCGVSYWGVVNSRQDGVPLKADFLAALLPLLCIPAVFSLFTGMYKW KDDDWKISRGVYLFVGMGVLLLLGAISAVIVTIRPWTVGVACLLVILFLVFAIGVIHY WTSNNFYLTRTQMLLVCSLAFLLALAAFLMGLFQEKPFVGASIGYFSFLFLLTGRALT VLLSPPIVVYSPRVLPVYVYDAHADSAKNVSYAFLILYGIALATEVWGVIASLILNPP FIGAAISAITLVIAFSFAVSRPCLTLKMLEDAVHFLSKDTVVQAMSRSANKTRNAISG TYSAPQRSASSAALLVGDPAITLDRAGNFVLPRADVMKLRDRLRNEEITAGSFFCGVK NCLMIGSPVDVDYRRNMCAHARILALEEAIDTEWVYMWDKFGGYLLLLLGLTAKAEQI QDEVRLRLFLDSIGLSDLSAKEIKKWMPEDRRHFELIQESYIREKEMEEEVLMQRREE EGKGRERRKALLEREERKWKELEISLLSSIPNAGSRDAAAMAAAVRAVGGDSALEDSF ARDRVSSIARHIRKAQLARRAEQTGIPDTVCILDDEPRSTGRHCGEIDLCLCESKKVS FSIAVMVQPVSGPVCLFGTEFQKKVCWEILVAGSEQGMEAGQVGLRLVTKGERMTTVA KEWNIGASSIADGRWHLVTVTIDADLGEATSFIDGVYDGYQNALPLPRNNGIWEPGTD IWVGARPPTDLDAFGRSDSEGSDSKMQIMDAFLWGRCLTEDEVAMLHTAICSAEYGLF DLAAEDAWHGSYSARVDDWESEEANFELYDQEDVEWDGQYSSGRKRHARDSVAIDIDS FARRPRKPRFETREEVNQRMLSVERAVREALIAKGERNFTDQEFPPDDRSLFVDPMNP SLKLQVVSEWMRPSDIAKEVSISSQPCLFSGSVNSSDVCQGRLGDCWFLSAVAVLTEM ARISEVIITPEYNEEGIYTVRFCIQGEWVAVVVDDWIPCESPGKPAFATSRKQNELWV SILEKAYAKLHGSYEALEGGLVQDALVDLTGGAGEEIDMRSPQAQIDLASGRLWSQLL HFKQEGFLLGAGSPSGSDAHISSSGIVQGHAYSILQVREVDGHKLVQIRNPWANEVEW NGPWSDSSQEWTERMKHKLKHVPQSKNGVFWMSWQDFQIHFRSIYVCRVYPPEMRYSV HGQWRGYSAGGCQDYDSWHQNPQYRLRVTGRDALYPVHVFITLTQGVGFSRKTNGFRN YQSSHDSSMFYIGMRILKTRGCRAAYNIYMHESVGGTDYVNSREISCELVLEPYPKGY TIVPTTIHPGEEAPFVLSVFTKAPIKLEAV</aaseq> |
| Gene Sequence |
<dnaseqindica>56..183#605..855#1334..1516#1613..1748#1891..2021#2097..2211#2295..3333#3418..3528#3782..3840#5115..5277#5676..5949#6044..6194#6280..6391#6492..6657#6775..6845#6988..7079#7235..7397#7648..7719#8053..8424#8505..8620#8715..9012#9334..9653#9846..10196#10498..10816#10937..11041#11162..11290#12515..12895#12981..13118#13463..13699#13778..14083#gtttgagcctgtatacatgatatcagcttccagaaggtgattgcagagggggaggatggaagaggaggaacaccgcggggttgttcttgtttgcagcatctgcggtttcctcttcgctgtccttggccctctcagcttttggatcctatgggctgtgaactggaggccgtggaggctatataggtatgctcgagattaggcgtgcttctgttggcatctagcatttgcatctgtagtctgcgaggttgcattattaatcatacgttatgtttccataaatttcagccagcagagtgcatacaacaatgtagatttggtatcattatcactctaacaactgaaattatttattgtgtctttattatgctgctaatccaacaccacaccttatatattttaagcctgccattcatatgtagattatggaatacgatcatgtaattgtgtctaactacaatcacaagtagtgccctgaaaaacgttgatgtttttctattcttatatcttctatgcatatttgtttatgttgatttgttcctgatgttctgagagctggattatctcattaagctactaacattttcgttcttgttttgttactcagttggatatatgcaaggaaatggccagcatatgtgcaaggacctcaactaagcacactgtgcagctttttcactctttttgcatggcttgtcgtcgtttctccaataacagttttgttagtatggggaggcatccttattgctcttttggaaaggaacataattgggctagctgttataatggtcggtgttgctttgcttctgtcattctactctataatgttgtggtggagaacgcaatggcaaagctccagtaagttttctactgtttacatgcttttattttctaggacattttgtgatgactactcaatgtgcttgtacttagggtcatggtttcattaaaccaaatattctgataagtggctgtttattcatgacttctaagtcatgggttcattaaatttgatattctaataggcgatgattgtctaggacttctctgcttattttgggggtccaatttgtttctagacaattgtaactaggaaatggtataaatggaacattttggcatactcaaaggaaaacttttaaagcttcacggaggattaaaaatactagtatgctactacaaatttgaacttttctcactttataaaactgcaatttctaactctttgattcatgtggtattttgtatgtgctgtttctgcaaaattgggccgctctatcatctcattgttctttattaatatcttactgatgttaacaaatgttggttgttgcagaggctgttgcttatcttctactcctggcagtcggtctactgtgcgcttatgagttttgtgctgtttatgttacaaccggtgctagtgcttctgagctcaattccccatctggcttcttctttggggtatctgcaatttcattggccataaatatgctttttataagtaaaatactgttcaatggtaaaattcttatccgttatctttatttgctgaatttcttatcacctgtaaataaaaaatttgagaaacataacttttcatacattacatgtacaggaagtggattcgatgttgatgagtatgtccgaaggttgtacaaatttgcatattctgattgtgttgaagtggcccctgtttcatgctcgcctgacccaccagatcctagtgagttatacatgacaaaatccagcaggtgacttcactaccttttcctctcttaaatactacataggcatatatgttccccaagtaacgtgcttgttctattatgctgttactggggtaagcaataatttctagtagttttggtaacatatcaatgccttaaaatgcagggtcttgcatctagggcttctttacctttgctcattgatggttcttgttgtctattccatcttatatggtctgacatcaaaagaagcacgctggctgggtgctttaacttcagttgcagtagtgattctcggtaatgatttggccttgtatctctaattgtgcatgatgtatctatttctttgacatcttattttcttttctacagactggaatttgggcctgtgttcatttaggtttgagcttcttaaaagtcggatgatagcgctatttgtagctggaacatcaagggttttccttatatgctttggggtgcactactggtttgtattcatctttatgcttttatagttgaacatatttatgatctagtactttgtgtttactaattaatgtgttacattaggtaccttggacattgcatcagttatgcttttgtagcatctgtgcttttagctgcagctgtctcttgctggctttctatttcaaacccctcagttgcaaggatagatgcattaagaagcacagtgataaaactccgagaaggttttcgaaggaaggggcaaactagttcttcaaactcatcagatggctgtggatctagtgtgaagcgtagtagtggcagtgttgaagctggtcctcatggcaatgctactgattctatgtacagaagcaactcacaaagcgattgtgtcaattggaacaatgttccttttgatcgatcgaatagttgtcaagaaggacagagctctgacaagaacatagatagtggacgtgccagtttagcccatcgcagtaattcgtgcctttctgctgtagctgtacaagaccctgaaactgctgtagtttcagcggacaggcatggggacccaactgcttcactcgttgtttgctctagtagtggtttggaaagtcaaggctgcgagtctagtggatcagctactgcctcgggtaatcagcagctattggatttgaacttggctgcaatttttcaggacagactgaatgatccaagaatcacatctatgctaaaaaggaatggtggactaggagatgtagaattggctaatcttcttcaggataaaggccttgatccaaatttttcctatatgatgaaagacaaggttatggacccacgaattttagctttgctgcagagaagcagcttagatgcagacagagagcatcaagatgatgtagatgtcactggtactgattcagataggttagatacaactattgcaaaccagatctcgctatcagaggaacttaggagaagtgggctagaaaattggctaaatctttcaaggctgatgtttcatcaagtagcaggttcaccaatacgtgcttttgttgtttttaccctaatattcatcatagaaacagttactgtggctgtccatcgaccgaagcccatcaaggttataaatgcaactcatgaacaggtaagtttgttgcaaagacagtataaaaactttgaaatgttgattgtgttgttaatcaagagtgtcctttttcgaatttggcagtttgagtttggtttctcgatactgcttctatcaccagttgtctgctccattatggcattcatttggtccctgtgtgcagaagagatgacgatgacatccaaacctcggaaggtgatgttgaacaaaagaaatatatttgtcgtattaataccaagcacttattttttaatcatggcaggacaagtccttttgcttcttcactagacatttagatatgttctgtcctccttaactcgtctacaaaatatcaaatctgattacaatgtgtgctttatcttactttgaccatgtagttgattttttttttcaaaatcttcgtggattttcagctatagtgacttttcctttcttttcccatctacagtatggcttcatcgcatggttgctgagtacttgtgttggtctacttctgtctttcttgaggtatacatgtttgttatttaatatttaaaacttcacttagttactactgcattttgccttgtgtaaatcctggagtactcatttgttttccttccctgctttggtgtcttgctggtttgactagaagtttcacatgggtatcaaatgatacatgcacagcataatagccatgtccacttgtttcgtatattctacacaagtacattttattgtataagtctgttactataatactgtgtctctgcctggttcatttgctacgccaccttgtgcttggaatctttttatattggtagaattgatcatgtgtgttattattagcattagtacgaataaatgtcatcgagtcattcttttgtttgttgcaaacaaaagagacctatctattttcatgaagataacctcaccacttgttatctgatcatggtacaattacttgttaagttctctatgcacagctcattttgagttgatgtacataaaattatactctgacgtgctttgcgctgctatatggtagtcttgtcttagacctcctttgggacagcttttagccagcttttgaacagaaaaaagctagcaaacttctcacacggctagctcatcccaggagttgctcacaaaaggaagcccattcatgaagacaaggagatctcatttccttgctcctccatggcttttgtgggatttgtttgagtaatttacccacctttgccatcatacccactgccagcgctcagcctgaaaactcttacttgatccccttccattccattagcctactcaaggtcctgatcatcgctggatccaccttgagctcctggttgccacggcctctaggtcccaatcgctaacgcagtaccgcctcaagctccctgtcgccgtcacctccacatcctagtcgccaccaccgccttgagctccccatctttgtcacctccaggtcttggtcaacaccaccatctagagttcctggtcactgctgcctctaagattccagtcatagctgctgcctcgagttcctagtatgcaaagttttagctttaagtggattactccttttatctggataaggtgtcctagcttgctaaaatgtgcttcttgattattctattgttcctgttttgacaattgacatattcacagcatcttgcatatgccttgtttatacatatgtatgactctctagccactagatttttataatcattgattcatttcattggggaagtacttacacttttgtctgaaatactacctctgccattctgcagcaagtcatcagttattttgggtttgtcactcacggtcccacttatggtggcttgtctctcatttgctattcccatatggatgcgtaacggttaccgtttctggattccgggaggggagcttgatagccgtgaaaatatccgccaagctccaggaaagaaagaggtttgttgtttttgttaattcagcagcaggacacccttttgtttatatatttggatatccaataaatgtctgtataagctatttgcgtagttgtaagggaagtactgtaaccagtatcatcttactatatggccttatgtcaatatactgttaacaacttctggatgaatgcctgaatatgtttactgaatcaattttcactgtattgttgtgattcctatcagcattagataaaatcttgttttgtaggctatcctactgaaaagaattcaggctctgttttacagcacatacctcttgacctctctaatgttattataaatagtgtagaacgcattcagaccttccagcattgttaatgattttcatgttactgatcgttgttatcttcttgtcagcgcgctctgtttgccatcagcataactgttttcactgcatcagtaattggccttggtgcaatagtatcagcaaaacctctagatgcattgggctacaagggatgggatgctgataaaaagagtttctattctccctatgcaacatcgatgtatcttggatgggcattatcatcaacgattgctgtacttgccactggggtgatacctattgtcgcttggtttgctacttatcgattttcaccttcatcagctatatgtgttggcctttttgcaagtatgttctactcttcttttccgttctcattgcatattaaatagagcacatcacacttttggataactggcacttgactggtgactttctgcagctgtccttgtatccttttgtggtgtgtcctactggggagtggtaaattcacgacaggatggagttcctctgaaggctgatttcctggcagctttacttccattactttgcattcctgctgtgttctcgttgtttactgggatgtataaatggtatggaaaattggaaaccatgtttattatttgtacgtccttgcatacttctgctcctctgagatgaaattgcaaataatttcaggaaggatgatgattggaagatttcccgtggagtttacctatttgttggcatgggagtgctgttattgcttggtgcaatatctgcagtaattgttacaattagaccttggactgtaagtaaagtttgtggcttggttttcaaaataaatttatcataatattcatttcatttgtgataagagacagcataattgatctgtcattgctatgcaggttggagttgcatgcctccttgtcatcttgttccttgtatttgctattggtgtcattcactactggacatctaacaatttctatttaacaaggacacaaatgttacttgtttgctcccttgcttttctccttgccttggctgctttcctgatgggcttatttcagggtgactacatcttcttatatgcttttcaacctgttattgctatttttctttctccgagttacataatagtttcccatattatgcacatagcttaaatcagctgatgattttttgcagagaaacctttcgttggggcatctataggttatttctcgtttctatttcttctcacaggaagagcattgactgtaagtttcatatcttttttcttaacttaagttagctacatgtttaagataatttctgttgaacatctcatgctagttttgctacttgtgagttatctcaagagttatgcttgtaatattgaatttactgtactcttctcaggtccttctatcaccaccaattgtcgtgtattcaccgagggtattacctgtttatgtttatgatgcccatgcagactctgctaaaaatgttaggtacggatatcaggatatggcctttctaattctacatgcttatatatctttcctttgaccaggattggtcataactcataacttctttcttctatgctatttttagaagtactattgtcatctgagcacacaccctaacattgctgtatttacagttatgccttccttatcttgtatgggattgcattggctactgaagtttggggtgtcattgccagtctaatattgaatccaccatttattggggctgctatttctgctatcactcttgtaattgccttcagttttgctgtttctcgaccatgcctgactctcaaggttagtatttctgtttatccaaaatcagcagattattgaaatgagctctctcttaggaaaatctaggatttgtcacccaaaaatatgcacttcagcacttgccagttgcaactccaccacatgtagccgctcacatcagtggcacattggttggcaaatcctgaagtgccaataccaatggcaataatacactcctttatttcctcttttttcttgatccttcttaaattaattgtgaaatttttgtcagatgctggaggacgctgttcactttctcagtaaggatacagttgtacaagcaatgtcacgttctgctaacaaagtaagtttgctgtatatttgtattattcagatattcaacatatccctgcgtaacataggtaaatgctgttcttgcttgcatacttttctgttaaacctgcctcagtaagcctaccacttagcatctagcgctgagatcagaactatgttttcttaatctagtgcatcaagttcaatcatgtatgttcccactccttgtcagccttgacacttacataagtgctgctggtgttatctactccaatatccaggaccagtacttatcaatgagattattatgactctgatttttattatttctgtattgttaatagtatgctttgtgtgcaatcagacgagaaatgctatttctgggacttactcggcgcctcagagatccgcaagttctgctgctcttttggttggagatcctgctattacgttggacagggctgggaattttgtgcttcctagggctgatgtcatgaagctgagagatcgtttaagaaatgaagaaattacggcaggatcgttcttttgtggagtgaagaattgtttaatgattggatcccctgtagatgtagattatcggaggaatatgtgtgcccatgcacgtattttggctttggaagaagcaattgatactgaatgggtttacatgtgggacaagtttggtggttatttacttcttctgcttggcttgactgccaaagctgagcaaatacaggtatgcatttgggactactcccacctaatcttttttgttcttaaaatctgtaacaatgttggaaatgtctgttgtttcaggatgaagttcgtcttagactttttctggacagcataggtctttctgacttaagtgccaaagaaatcaagaaatggatgcctgaagatcgaagacattttgaacttattcaagaaaggttattgttaaaacttttatgtatgcaatgtttctatctgcaaccctgtaaacctctgtgcccgccatctaaattctcttattcgcttattcagctacataagggaaaaagaaatggaagaggaggtcttgatgcaaaggcgcgaggaagaagggaagggaagggaaaggaggaaggcattgttggagagagaggaacgaaaatggaaggagcttgaaatatcactgctatcttctattcctaatgctggaagcagggatgctgcagccatggcagcagctgtcagagctgtgggtggtgattctgctctggaagattcttttgcaagggacagggtctcgtcaatagctcgtcacataagaaaggcacaattggctcggcgagcagagcaggttctgttactgttcctgtaatcattagattttatcaacttctaaagcacaccttaatgatgatatagttctcatggtgtaccttttgagtgcatgaactgaacatattcatctttggatatttattatctgtttttgccatccattttggagttggttatctgacaaagcatcaatttttgcatgacagttagttacgtgtaatagcctggaatcatttcttctgtttataaatttagtaccatatgttatacatgcagttcttgtaattacttttttactgatgctctttggatcttttacttgttacgaactgtgcagactggcattccagacactgtttgcatacttgatgatgaaccaaggagtactgggcgtcattgtggagagattgatttgtgtctttgtgaaagtaaaaaagttagcttctccattgcagtaatggtgcagccggtgtcaggtccagtctgtctttttggaacagaattccagaaaaaagtgtgctgggaaatcttggtggcagggtcagaacagggaatggaagctggccaagttggtcttcgattggtgactaagggtgaaaggatgacaaccgttgcaaaagagtggaatattggtgcatcaagtattgctg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