The nucleic acid data:
IRESite Id: 369 Version: 1
Originaly submitted by: Václav Vopálenský
Reviewed by: Martin Mokrejš Last change: 2008-06-10 09:28:24
IRESite record type:
  negative_control_plasmid_with_promoter_and_without_putative_IRES
The shape of the nucleic acid molecule translated:
  linear
The quality of the mRNA/+RNA sequence:
  end-to-end_full-length_mRNA
The mRNA/+RNA description: 
Bicistronic mRNA containing DsRED2 and EGFP. The sequence ends at its 3'-end right after the poly(A) signal
from SV40 mRNA and thus the 3'-UTR might be slightly wrong.
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:
pRG
The name of the promoter used to express this mRNA:
  CMV_IE
Description of the plasmid (facultative for promoter-less plasmid records):
The mammalian bicistronic vector pRG is a derivative of the pDsRed2-C1 vector containing the enhanced green fluorescent protein (EGFP) gene from the pEGFP-N1 vector (Clontech) inserted into the HincII site. A new stop codon was introduced at the end of the DsRed2 gene by ligation of the 5'-GATCCAGCAAGATATCCTAA-3'/5'- GATCTTAGGATATCTTGCTG-3' adaptor into the BglII site. IRESite notes about verification of plasmid sequence: The complete sequence of intercistronic region was determined by sequencing using DsRed1-C Sequencing Primer (Clontech; #6483-1; 5'-AGCTGGACATCACCTCCCACAACG-3'), nt 1205–1228.
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
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:
pRG.jpg
Plasmid sequence verified (partially/completely) by IRESite (more details in Remarks):
  plasmid sequence confirmed by IRESite curators by restriction analysis + parts by PCR + sequencing
A promoter reported in cDNA corresponding to IRES sequence:
  not tested
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: Václav Vopálenský Reviewed by: Martin Mokrejš
The abbreviated name of this ORF/gene:
DsRED2
The description of the protein encoded in this ORF:
red fluorescent protein (DsRed2 version, Clontech)
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:
  not tested
The ORF absolute position (the base range includes START and STOP codons or their equivalents):
  31-726
ORF
ORF position:   2
Version: 0
Originaly submitted by: Václav Vopálenský Reviewed by: Martin Mokrejš
The abbreviated name of this ORF/gene:
EGFP
The description of the protein encoded in this ORF:
Red-shifted variant of wild-type GFP optimized for brighter fluorescence and higher expression in mammalian cells (Clontech).
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:
  not tested
The ORF absolute position (the base range includes START and STOP codons or their equivalents):
  796-1515
Remarks:
The plasmid was used as the negative control for experiments in articles:
Hepatitis C virus internal ribosome entry site initiates protein synthesis at the authentic initiation codon
in yeast. (Masek et al., 2007; PMID: 17554033)
Firefly luciferase gene contains a cryptic promoter (Vopalensky et al., 2008; PMID:).
Citations:
Masek T., Vopalensky V., Horvath O., Vortelova L., Feketova Z., Pospisek M. (2007) Hepatitis C virus internal ribosome entry site initiates protein synthesis at the authentic initiation codon in yeast. J. Gen. Virol. 88(Pt 7):1992-2002
Last change to the database: 2015-04-16 16:45:23 GMT+1