The nucleic acid data:
IRESite Id: 121 Version: 6
Originaly submitted by: Martin Mokrejš
Reviewed by: Martin Mokrejš Last change: 2006-10-30 00:00:00
IRESite record type:
  plasmid_with_promoter_and_putative_IRES_without_translational_characterization
The shape of the nucleic acid molecule translated:
  linear
The quality of the mRNA/+RNA sequence:
  hopefully_full-length_mRNA
The mRNA/+RNA description: 
Monocistronic mRNA containing 5'-UTR from BCL2 and firefly luciferase with 6 extra N-terminal codons from
BCL2.
The mRNA/+RNA sequence represented in the +DNA notation:


Credibility of mRNA sequence:
  end-to-end_sequence_completely_same_as_in_the_experiment
The name of the plasmid:
pBCL2-FL-polyA
The name of the promoter used to express this mRNA:
  T7
The in vivo produced transcripts are heterogeneous (due to any of promoter?/splicing?/cleavage?/breakage?):
  not tested
The in vivo produced heterogeneous transcripts occur due to alternative splicing:
  not tested
A promoter reported in cDNA corresponding to IRES sequence:
  not tested
The abbreviated name of the donor gene or virus from which this IRES was excised and inserted into the plasmid:
BCL2
The origin of IRES in the plasmid:
  nuclear
The donor organism of the IRES segment:
Homo sapiens HeLa (ATCC CCL-2)
The DNA sequence of the plasmid in (+) orientation annotated by its secondary structure:


GenBank formatted file with annotated plasmid sequence hyperlinked from vector image map:
pBCL2-FL-polyA.jpg
The total number of notable open-reading frames (ORFs):
  1
Notable Open-Reading Frames (ORFs; protein coding regions) in the mRNA/+RNA sequence:
ORF
ORF position:   1
Version: 1 Last change: 2006-06-05 00:00:00
Originaly submitted by: Martin Mokrejš Reviewed by: Martin Mokrejš
The abbreviated name of this ORF/gene:
FLuc-fusion
The description of the protein encoded in this ORF:
firefly luciferase with 6 extra N-terminal codons from BCL2
The translational frameshift (ribosome slippage) involved:
  0
The ribosome read-through involved:
  no
The alternative forms of this protein occur by the alternative initiation of translation:
  no
The ORF absolute position (the base range includes START and STOP codons or their equivalents):
  1151-2821
Remarks:
Annotation of the plasmid: 16-812: CMV immediate early enhancer/promoter; 751: CMV transcription start;
869-1005: chimeric intron; 977-1001: FWD primer; 1049-1067: T7 RNA polymerase promoter (-17 - +2); 1066: T7
promoter transcription start (+1); 1078-2215: BCL2 5'-UTR; 2216-3886: FFluc-fusion protein ORF; 4794-2139:
SV40 enhancer; 4983-5118: SV40 ori; 5200-5990: Kan/Neo resistance (neomycin phosphotransferase); 8208-8232:
REV primer

In vitro transcripts from this plasmid were used in Fig. 2. Otherwise pretty much useless construct showing
only that phpBCL2-FL in contrast to this plasmid is less translated because of the "php" stem loop barrier.
Fig. 4B shows that translation of capped and polyA tailed monocistronic mRNA is about 11x higher than that of
uncapped mRNA and that "php" stem loop as the barrier decreases translation down to about 8%.
Citations:
Sherrill K. W., Byrd M. P., Van Eden M. E., Lloyd R. E. (2004) BCL-2 translation is mediated via internal ribosome entry during cell stress. J. Biol. Chem. 279(28):29066-29074
IRESs:
IRES:
Version: 2 Last change: 2006-06-05 00:00:00
Originaly submitted by: Martin Mokrejš Reviewed by: Martin Mokrejš
The IRES name:
  BCL2
The functional status of IRES:
  functional
The IRES absolute position (the range includes START and STOP codons or their equivalents):
  191-1328
How IRES boundaries were determined:
experimentally_determined
The sequence of IRES region aligned to its secondary structure (if available):


Citations:
Sherrill K. W., Byrd M. P., Van Eden M. E., Lloyd R. E. (2004) BCL-2 translation is mediated via internal ribosome entry during cell stress. J. Biol. Chem. 279(28):29066-29074
Last change to the database: 2015-04-16 16:45:23 GMT+1