The nucleic acid data:
IRESite Id: 126 Version: 2
Originaly submitted by: Tomáš Mašek
Reviewed by: Tomáš Mašek Last change: 2008-07-04 13:02:03
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 control plasmid pFGAL4-L428 which comprises luciferase and Gal4 ORFs. L428 clone
was selected from pFGAL4::lambda library containing randomly cleaved phage lambda DNA for its comparable
length  of intercistronic region to tested pFGAL4h-HCV1.
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 plasmid:
pFGAL4-L428
The name of the promoter used to express this mRNA:
  TPI
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:
pFGAL4-L428.jpg
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:
ORF
ORF position:   1
Version: 0
Originaly submitted by: Tomáš Mašek Reviewed by: Tomáš Mašek
The abbreviated name of this ORF/gene:
Fluc
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:
  no
The ORF absolute position (the base range includes START and STOP codons or their equivalents):
  1-1653
ORF
ORF position:   2
Version: 0
Originaly submitted by: Tomáš Mašek Reviewed by: Tomáš Mašek
The abbreviated name of this ORF/gene:
GAL4
The description of the protein encoded in this ORF:
DNA-binding transcription factor 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):
  2082-4727
OPTIONAL: The function of the encoded protein
Molecular Function: transcriptional activator activity (TAS)
Biological Process: galactose metabolism (TAS) regulation of transcription, DNA-dependent (TAS)
Cellular Component: nucleus (TAS)
Remarks:
L428 sequence was inserted into intercistronic position of pFGAL4 bicistronic plasmid to create bicistronic
control vector that can be used in the pFGAL4h/PJ69A reporter system. 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.

Vector was created by IRESite curator - thus whole sequence is verified by above mentioned methods.
Citations:
Masek T., Vopalensky V., Horvath O., Vortelova L., Feketova Z., Pospisek M. (2007) Hepatitis C virus internal ribosome entry site initiates protein synthesis at the authentic initiation codon in yeast. J. Gen. Virol. 88(Pt 7):1992-2002
Last change to the database: 2015-04-16 16:45:23 GMT+1