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: both_UTRs_incomplete
The mRNA/+RNA description:
mRNA produced by bicistronic plasmid pFGAL4h which comprises luciferase and Gal4 genes as the first and
the second cistron, respectively. 13G:C hair pin loop was inserted between the two cistrons to prevent
The mRNA/+RNA sequence represented in the +DNA notation:
Credibility of mRNA sequence: end-to-end_sequence_completely_same_as_in_the_experiment
The description of the protein encoded in this ORF: GAL4 is a DNA-binding transcription factor. Gal4p is required for the activation of the GAL genes in response
to galactose; repressed by Gal80p and activated by Gal3p.
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: no
The ORF absolute position (the base range includes START and STOP codons or their equivalents): 1734-4379
pFGAL4h bicistronic plasmid has been designed for the use in the pFGAL4h/PJ69A reporter system. pFGAL4h vector
bears genes for the firefly luciferase and Gal4p as a first and second cistron, respectively.
A stable stem-loop hairpin containing 13G:C pairs was inserted between the two genes in order to prevent
ribosome read-through from the first to the second cistron. The yeast strain PJ69A contains besides the GAL4
and GAL80 deletions also reporter genes coding for the yeast Ade2 and His3 proteins and for the bacterial
beta-galactosidase - all of them under the control of the Gal4-inducible Gal1 promoter. Thus pFGAL4/PJ69A
represents a specialised and sensitive system,
which allows an enhancement of the measured signal by in vivo coupled transcription and enzymatic detection.
This system can be used for both measuring the ratio of the beta-gal/luc enzymatic activities and searching
for IRES activity by screening the colony growth rates on selection media lacking adenine and histidine and
containing various concentrations of the competitive inhibitor of the histidine biosynthetic pathway.