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GenBank: W43399.1
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LOCUS W43399 533 bp mRNA linear EST 28-JAN-2011 DEFINITION 22792 CD4-15 Arabidopsis thaliana cDNA clone G11E11T7, mRNA sequence. ACCESSION W43399 VERSION W43399.1 DBLINK BioSample: SAMN00155038 KEYWORDS EST. SOURCE Arabidopsis thaliana (thale cress) ORGANISM Arabidopsis thaliana Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta; Spermatophyta; Magnoliopsida; eudicotyledons; Gunneridae; Pentapetalae; rosids; malvids; Brassicales; Brassicaceae; Camelineae; Arabidopsis. REFERENCE 1 (bases 1 to 533) AUTHORS Newman,T., deBruijn,F.J., Green,P., Keegstra,K., Kende,H., McIntosh,L., Ohlrogge,J., Raikhel,N., Somerville,S., Thomashow,M., Retzel,E. and Somerville,C. TITLE Genes galore: a summary of methods for accessing results from large-scale partial sequencing of anonymous Arabidopsis cDNA clones JOURNAL Plant Physiol. 106, 1241-1255 (1994) PUBMED 7846151 COMMENT On Jan 5, 1998 this sequence version replaced gi:1327883. Contact: Thomas Newman MSU-DOE Plant Research Laboratory Michigan State University MSU-DOE-PRL, Michigan State University,Plant Biology Bldg.,E. Lansing,Mi Tel: 517-353-0854 Fax: 517-353-9168 Email: 22313tcn@ibm.cl.msu.edu Seq primer: T7. FEATURES Location/Qualifiers source 1..533 /organism="Arabidopsis thaliana" /mol_type="mRNA" /db_xref="taxon:3702" /clone="G11E11T7" /tissue_type="seedling hypocotyl" /clone_lib="SAMN00155038 CD4-15" /dev_stage="3 day-old" /ecotype="Columbia" /note="Vector: pBluescript SK-; Site_1: EcoRI; Site_2: EcoRI; Using 5 ug of polyadenylated mRNA from 3 day-old Arabidopsis thaliana (Columbia) seedling hypocotyls as template and oligo d(t) as primer, first strand synthesis was catalyzed by Moloney murine leukemia virus reverse transcriptase (Pharmacia). Second-strand cDNA was made using the procedure of Gubler and Hoffman (1983) except that DNA ligase was omitted. After the second strand reaction, the ends of the cDNA were made blunt with Klenow fragment and EcoRI/NotI adapters (Pharmacia) were ligated to each end. The cDNA was purified from unligated adapters by spun-column chromatography using sephacryl s-300 and size-fractionated on a 1% low melting point mini-gel. Size selected cDNAs (2 - 3 kb) were removed from the gel using agarase (New England Biolabs), phenol:choloroform extracted and precipitated using 0.3 M NaOAc (pH 7)/ethanol. A portion of each cDNA size-fraction (0.1 ug) was co-precipitated with 1 ug of lambdaZapII (StratageneEcoRI digested, dephosphorylated arms and then ligated in a volume of 4 ul overnight. Each ligation mix was packaged in vitro using Gigapack II gold packaging extract (Stratagene). We have determined that although first strand cDNA synthesis was initiated using dT, almost all of the cDNAs begin 8-10 bp from the poly-A tail. The reason for the loss of the poly-A tail is most likely due to lower than anticipated nucleotide levels during the Klenow repair of ragged ends before the addition of linkers (3'-5' exo instead of 5'-3' pol). When this library is used please reference the ABRC and: Kieber, J. et al. (1993) Cell 72:427-441." ORIGIN 1 cttgggccat gggcctatca cttaactttc atacaaagga catccagtga tcactatttg 61 agtttttgac attgtgtgat acatttgaag gaaaggagta atgtttaagg ttgagttaac 121 atgtaagaga gtctgtggag ttactcagtt cagtagggaa gaggaagctt cgtctgtctg 181 ctgctgctcg atgtccaagt tcagctgcgg cgggccagct aaactcagta tgacaggtaa 241 gaaaacgagc ccatgcaaga agccgattat aaccaaagcc aagtacattt ggaagtaata 301 caccacaaat atctcagatc ttgcaaagca aagcacggtc actccaacaa gttttgtgag 361 tgtgatccca ctggaaactg aagctcccat ggtttccagt gcctccctcg ntccaatggt 421 ccctgtcccc actaatcatc agggaagcat ggtgatnttg tgcacgcgga ttctacagca 481 atcccggntg ncattatcaa ttcccaacag gaactggatt gggtnggggg cgc //
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