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24399 CD4-13 Arabidopsis thaliana cDNA clone E2A6T7, mRNA sequence

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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