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 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:
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:
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:).