IRESite record type: plasmid_with_promoter_and_putative_IRES_translationally_characterized
The shape of the nucleic acid molecule translated: linear
The quality of the mRNA/+RNA sequence: both_UTRs_incomplete
The mRNA/+RNA description:
Part of the T7-derived transcript studied in in vitro assays. The 5'-UTR could have been longer but do not
know the necessary details. The intercistronic region contains 5-UTR sequence of EMCV-R virus (bp 315-843).
The mRNA/+RNA sequence represented in the +DNA notation:
Warning: available plasmid sequence is exactly same as the mRNA sequence. It seems we never managed to reconstruct the original plasmid sequence used to express the studied mRNA.
Credibility of mRNA sequence: reverse_engineered_fragment_and_the_rest_is_a_guess
The description of the protein encoded in this ORF: beta-glucuronidase
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): 1316-3127
Remarks:
The intercistronic region inherited quite a long sequence from parental plasmids (64bp). The EMCV-R fragment
was inserted downstream of this spacer using EcoRI and Nco sites just in front with the GUS ORF.
The IRES absolute position (the range includes START and STOP codons or their equivalents): 785-1315
How IRES boundaries were determined: experimentally_determined
5'-end of IRES relative to last base of the STOP codon of the upstream ORF: 65
3'-end of IRES relative to last base of the STOP codon of the upstream ORF: 595
5'-end of IRES relative to first base of the START codon of the downstream ORF: -531
3'-end of IRES relative to first base of the START codon of the downstream ORF: -1
The sequence of IRES region aligned to its secondary structure (if available):
Remarks:
The GFP-IRES-GUS cassette was also cloned into yeast vector PyeDP1/8-2 and expression of reporters studied in
unnamed yeast of strain 2805 grown at 30 oC! The EMCV-R IRES was found active about 10x above some negative
control.
The relative translation efficiency in % of this IRES: 100.000
Name of the plasmid used as the negative control. H-GFP-TMV_UI-GUS
IRESite Id of the plasmid used as negative control. 611
The relative translation efficiency in % of the negative control: 0.860
The size (length) of intercistronic region in the negative control: 78
The effect of 5'-cap analogs on translation: not tested
Rapamycin affects translation: not tested
Type of RNA subject to translation: endogenous_cytoplasmic_uncapped_T7_transcript_without_polyA_tail
Remarks:
Data from Figure 2B. The cells were infected by recombinant vaccinia virus. Also GFP-TMV_UI-GUS (TMV UI 5'-UTR
omega leader) was used as a negative control (IRESiteID:611).