The nucleic acid data:
IRESite Id: 326 Version: 1
Originaly submitted by: Tomáš Mašek
Reviewed by: Tomáš Mašek Last change: 2008-06-04 22:06:57
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:
  3UTR_incomplete
The mRNA/+RNA description: 
Putative bicistronic mRNA transcript derived from pBSRLucEFluc plasmid through SV40 promoter. The plasmid has
in the intercistronic region T7 promoter.
The mRNA/+RNA sequence represented in the +DNA notation:


Credibility of mRNA sequence:
  reverse_engineered_sequence_and_should_match_experiment_maybe_except_3UTR
The name of the plasmid:
pBSFLucERLuc
The name of the promoter used to express this mRNA:
  SV40
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:
EMCV-R
The origin of IRES in the plasmid:
  viral
The donor organism of the IRES segment:
Encephalomyocarditis virus Rueckert
The DNA sequence of the plasmid in (+) orientation annotated by its secondary structure:


Plasmid sequence verified (partially/completely) by IRESite (more details in Remarks):
  plasmid sequence confirmed by IRESite curators by restriction analysis + parts by PCR + sequencing
GenBank formatted file with annotated plasmid sequence hyperlinked from vector image map:
pBSFLucERLuc.jpg
The total number of notable open-reading frames (ORFs):
  2
Notable Open-Reading Frames (ORFs; protein coding regions) in the mRNA/+RNA sequence:
ORF
ORF position:   1
Version: 0
Originaly submitted by: Tomáš Mašek Reviewed by: Tomáš Mašek
The abbreviated name of this ORF/gene:
FLuc
The description of the protein encoded in this ORF:
Firefly luciferase
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):
  269-1921
ORF
ORF position:   2
Version: 0
Originaly submitted by: Tomáš Mašek Reviewed by: Tomáš Mašek
The abbreviated name of this ORF/gene:
RLuc
The description of the protein encoded in this ORF:
Renilla luciferase
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):
  2636-3571
Remarks:
Plasmid DNA was digested by EcoRV/SalI and BamHI/SalI giving fragments 4321/1965 bp and 3392/1877(17) bp,
respectively. Amplification with Firefly/Renilla gene specific primers (Fluc_seq, gTg gAC gAA gTA CCg AAA gg,
position within plasmid sequence:2118, Rluc_revseq, CTGCATGTTTTTCTGAATC, position within plasmid
sequence:3060) gave one fragment of expected size. The sequence of promoter and intercistronic regions was
determined by sequencing (regions 54-626 and 2180-2958 of plasmid sequence). More detailed data are annotated
in GenBank sequence file.
Citations:
Hennecke M., Kwissa M., Metzger K., Oumard A., Kroger A., Schirmbeck R., Reimann J., Hauser H. (2001) Composition and arrangement of genes define the strength of IRES-driven translation in bicistronic mRNAs. Nucleic. Acids Res. 29(16):3327-3334
IRESs:
IRES:
Version: 2 Last change: 2011-04-08 22:02:13
Originaly submitted by: Tomáš Mašek Reviewed by: Tomáš Mašek
The IRES name:
  EMCV-R_272-831
The functional status of IRES:
  functional
The IRES absolute position (the range includes START and STOP codons or their equivalents):
  2072-2633
How IRES boundaries were determined:
experimentally_determined
5'-end of IRES relative to last base of the STOP codon of the upstream ORF:
  151
3'-end of IRES relative to last base of the STOP codon of the upstream ORF:
  712
5'-end of IRES relative to first base of the START codon of the downstream ORF:
  -564
3'-end of IRES relative to first base of the START codon of the downstream ORF:
  -3
The sequence of IRES region aligned to its secondary structure (if available):


Citations:
Hennecke M., Kwissa M., Metzger K., Oumard A., Kroger A., Schirmbeck R., Reimann J., Hauser H. (2001) Composition and arrangement of genes define the strength of IRES-driven translation in bicistronic mRNAs. Nucleic. Acids Res. 29(16):3327-3334
Last change to the database: 2015-04-16 16:45:23 GMT+1