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: 3UTR_possibly_incomplete
The mRNA/+RNA description:
Putative in vivo CMV promoter-derived transcript produced from bicistronic plasmid
hp-pbetaGAL/HIAP2(1073-1222)/CAT which comprises beta galactosidase and chloramphenicol acetyltransferase as
the first and the second cistron respectively and the part of human c-IAP1 5' UTR (nt from -150 to nt -1 of
the original sequence) mRNA mRNA cloned between them, with a hairpin in front of the first cistron.
The sequence ends at its 3'-end right after the poly(A) signal from BGH mRNA and thus the 3'-UTR might be
The mRNA/+RNA sequence represented in the +DNA notation:
Credibility of mRNA sequence: reverse_engineered_sequence_and_should_match_experiment_except_3UTR
The name of the promoter used to express this mRNA: CMV
Description of the plasmid (facultative for promoter-less plasmid records): Plasmid containing part of human c-IAP1 5' UTR (nt from -150 to nt -1 of the original sequence) inserted
between beta galactosidase and chloramphenicol acetyltransferase reporter genes. A synthetic DNA fragment was
inserted into the bicistronic constructs to introduce a stable stem loop structure 72 nt downstream from CMV
transcription start site.
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: c-IAP1
The relative translation efficiency in % of this IRES: 100.000
Name of the plasmid used as the negative control. hp-pbetaGAL/CAT
IRESite Id of the plasmid used as negative control. 521
The relative translation efficiency in % of the negative control: 13.100
The size (length) of intercistronic region in the negative control: 337
The effect of 5'-cap analogs on translation: not tested
Rapamycin affects translation: not tested
Type of RNA subject to translation: endogenous_nuclear_RNA_Pol_II_transcript
Translational efficiency was determined in thapsigargin (resuspended in DMSO)-treated cells.
DMSO-alone treated cells have about 10 times higher CAT production from hp-pbetaGAL/HIAP2/CAT plasmid compared
with empty hp-pbetaGAL/CAT vector.
Translational data are derived from Fig. 2C.
When rabbit reticulocyte lysates were primed with the hairpin-containing c-IAP1 IRES reporter construct
(hp-pbetaGAL/HIAP2(1073-1222)/CAT) and either the FLAGp97- or FLAG-p86-expressing plasmids, the addition of
p86 fragment resulted in the activation of c-IAP1 IRES (approximately 900 times higher production of CAT
protein, compared with the bicistronic construct that lacks the 5' UTR insert (hp-pbetaGAL/CAT)). See Figure