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GenBank: AA041133.1
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LOCUS AA041133 692 bp mRNA linear EST 28-JAN-2011 DEFINITION 24399 CD4-13 Arabidopsis thaliana cDNA clone E2A6T7, mRNA sequence. ACCESSION AA041133 VERSION AA041133.1 DBLINK BioSample: SAMN00155036 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 692) 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 Sep 19, 1997 this sequence version replaced gi:1517370. 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..692 /organism="Arabidopsis thaliana" /mol_type="mRNA" /db_xref="taxon:3702" /clone="E2A6T7" /tissue_type="seedling hypocotyl" /clone_lib="SAMN00155036 CD4-13" /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 (0.5 - 1 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 ctttgaactt agtcaccttc gatgagacga caatgcctgt acccatgtgt tcgtgcacag 61 gctctacgcg tcagtgttac aaatggggaa atggcggctg gcaatcgtct tgctgcacaa 121 ccaccttatc ccagtatcca ttaccgcaga tgccaaacaa gaggcattct cggatgggcg 181 gtaggaaaat gagcggaaac gttttctcga gattactcag ccgtttatca gcagaaggat 241 atgatctctc gtgtcctgtt gatcttaaag actattgggc taggcacggg acaaaccgct 301 acatcactat caagtagcct ctgttgttta cacagtccaa gtatgtttct ctcggcttct 361 taagattagg tgttggcttt gcctttattg gccaaagttt gtcctttggt tgtgnatggg 421 aaaaccattg gtaatgggga accttttacc aaaggttttt ggncccccag ggncaagtga 481 ccttccaaat cctttgttgg ggnagggant ncatgatgac angacctttg ggcctttcgg 541 taccaccaaa gngggggcaa ccaacccaan catttttgna cnncggncaa nggggttnaa 601 ggnggcaaag gcgnnaaaaa aaaacccccc cggttggggg accngggggc nggnccnang 661 gnggcccccc aaagggaaan ggccccnnaa aa //
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