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: end-to-end_full-length_mRNA
The mRNA/+RNA description:
In vitro T3 run-off transcript containing 5' region of PV (Mahoney strain) containing the IRES (-660 to -1)
plus the region encoding the first 37 amino acids of the viral nucleocapsid cloned upstream of the luciferase
reporter RNA bearing a poly(A) tail.
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: T3
Description of the plasmid (facultative for promoter-less plasmid records): Plasmid containing the major part of PV 5' UTR (-660 to -1) plus the region encoding the first 37 amino acids
of the viral polyprotein (+1 to +111) and firefly luciferase coding region (luc version of the luciferase
gene) with the poly(A)tract.
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: PV
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: not tested
The ORF absolute position (the base range includes START and STOP codons or their equivalents): 811-2463
IRESite notes about verification of plasmid sequence:
The complete sequence of IRES element was determined by sequencing using M13_reverse (CAGGAAACAGCTATGAC) and
Fluc_seq_reverse (AGGAACCAGGGCGTATCTC) primers.
The translation method used to study IRES function: in vitro
The in vitro translation system: HeLa cell lysate
The organism used for translation:
The temperature (in degrees of Celsia): 37
The relative translation efficiency in % of this IRES: 100.000
Name of the plasmid used as the negative control. pT3Luc-p(A)
IRESite Id of the plasmid used as negative control. 374
The relative translation efficiency in % of the negative control: 2.800
The effect of 5'-cap analogs on translation: no
Rapamycin affects translation: not tested
Type of RNA subject to translation: exogenous_RNA_with_ApppG_cap_without_polyA_tail
Translation of PV.IRES-pA RNA is unaffected by increasing amounts of m7GpppG competitor.
The translation of PV.IRES-pA RNA is about 10-fold more efficient than the translation of the
non-polyadenylated counterpart PV.IRES RNA.
Translational data are derived from Fig. 4A.