LOCUS GO549392 555 bp mRNA linear EST 14-APR-2009 DEFINITION Mdltb7524E13_e25440.g1 Apple_EST_Mdltb Malus domestica cDNA 5' similar to pir|F86447 F5D14.6 protein - Arabidopsis thaliana gb|AAF81326.1|AC007767_6 Strong similarity to an unknown protein F19D11.4 gi|7485759 from Arabidopsis thaliana BAC F19D11 gb|AC005310. EST gb|AV535485 comes from this gene, mRNA sequence. ACCESSION GO549392 VERSION GO549392.1 DBLINK BioSample: SAMN00167510 KEYWORDS EST. SOURCE Malus domestica (apple) ORGANISM Malus domestica Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta; Spermatophyta; Magnoliopsida; eudicotyledons; Gunneridae; Pentapetalae; rosids; fabids; Rosales; Rosaceae; Amygdaloideae; Maleae; Malus. REFERENCE 1 (bases 1 to 555) AUTHORS Korban,S., Vodkin,L., Liu,L., Gasic,K., Gonzales,O., Hernandez,A., Aldwinckle,H., Malnoy,M., Carroll,N., Goldsbrough,P., Orvis,K., Clifton,S., Pape,D., Marra,M., Hillier,L., Martin,J., Wylie,T., Dante,M., Theising,B., Bowers,Y., Gibbons,M., Ritter,E., Ronko,I., Tsagareishvili,R., Kennedy,S., Waterston,R. and Wilson,R. TITLE Apple Functional Genomics grant - NSF 0321702 JOURNAL Unpublished COMMENT Contact: Schuyler S. Korban Apple Functional Genomics grant - NSF 0321702 Washington University School of Medicine 4444 Forest Park Parkway, Box 8501, St. Louis, MO 63108, USA Tel: 314 286 1800 Fax: 314 286 1810 Email: submissions@watson.wustl.edu Library material provided by M.Wisniewski/C.Bassett/ J. Norelli/T.Artlip Library constructed by K. Gasic/S. Korban Library sequenced by Washington University Genome Sequencing Center original QR value of 815 was extended to value 816 (,) This trace has been recalled with phred original value before phred recall for SR was 816 COMM possible reversed clone; protein similarity on positive strand Seq primer: -40UP. FEATURES Location/Qualifiers source 1..555 /organism="Malus domestica" /mol_type="mRNA" /cultivar="Royal Gala" /db_xref="taxon:3750" /tissue_type="bark tissue" /clone_lib="SAMN00167510 Apple_EST_Mdltb" /dev_stage="plants subjected to 5 degrees C. for 24hrs" /lab_host="DH10B cells" /note="Vector: pBluescript II SK (+); Site_1: Not I; Site_2: EcoRI; Total RNA was extracted separately from bark tissue, using the method described by Gasic et al.(2004). Poly(A)+mRNA was isolated twice from total RNA, using the Oligotex Direct mRNA kit(Qiagen). mRNA was reverse transcribed into double stranded cDNA using a modified oligo18(dT) primer with an identifying tag sequence. [Tag identification when sequencing from 5'end: insert 18(A) TGCGAG]Double stranded cDNAs were size selected (more then 500bp), adaptored with EcoRI adapters at both ends and then digested with NotI. The cDNAs were then directionally cloned into EcoRI-NotI digested pBSII SK(+) phagemid vector(Stratagene). The total number of white colony forming units (cfu) was 6.0x10e6 cfu. The background of empty clones was less than 5%. Inserts ranged from 0.8 kb to 12kb, as determined by PCR." BASE COUNT 163 a 113 c 93 g 186 t ORIGIN 1 gctcgcattt tttttttttt tttttaaaaa gtaaaaggcc aatagtatat tcaatttttg 61 ccaacattgc aattctcagt attgtatttt gatttcatat cagatacaac tgcatagtag 121 tgacattcga gagcttgccc attcgggttt gacacaacaa tagaacatta tcccactaag 181 tggagtcggc tatagaaatc ttagaatgac aaccaaattg ctttgccttc tcatatattg 241 actcatttgc caaacagaca gaagaaaacc caaaatgttt gtaatcatgt accaaagcag 301 caatccagga gccctcatgc tagttaacat catacatatg caagaacgca ttatccttcc 361 tggctgaaaa tcattttggt ggtgccatat tccatttatt tgatgcttcc ttgtgcttct 421 tgtcttgata tagggataac gcgatggcgc ggcccttcga acatccaatt atcttcttca 481 agggatttaa ccaccttgtt aagtgcatcc ataagatcgg attttataac aagtaatgct 541 tcttgtttgc gatct //