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' UTR of human BiP (nt from -219 to -3) mRNA cloned upstream of the
luciferase reporter RNA lacking 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 5' UTR of human BiP (nt from -219 to -3) 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: BiP
The DNA sequence of the plasmid in (+) orientation annotated by its secondary structure:
Plasmid sequence verified (partially/completely) by IRESite (more details in Remarks): plasmid sequence confirmed by IRESite curators by restriction analysis + parts by PCR + sequencing
GenBank formatted file with annotated plasmid sequence hyperlinked from vector image map:
The total number of notable open-reading frames (ORFs): 1
Notable Open-Reading Frames (ORFs; protein coding regions) in the mRNA/+RNA sequence:
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): 309-1964
Remarks:
IRESite notes about verification of plasmid sequence:
The complete sequence of IRES element was determined by sequencing using Fluc_seq_reverse
(AGGAACCAGGGCGTATCTC) primer.
For detailed verification of whole plasmid sequence see item IRESite Id: 413.
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: 15.700
Name of IRES used as the positive control: BiP
Name of the plasmid used as the positive control. pBL-p(A)
Name of the plasmid used as the negative control. pBiL
IRESite Id of the plasmid used as positive control. 401
IRESite Id of the plasmid used as negative control. 396
The relative translation efficiency in % of the positive control: 100.000
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
Remarks:
The translation of BiP.IRES RNA is about 6-fold less efficient than the translation of the polyadenylated
counterpart BiP.IRES RNA (IRESite ID: 396).
Translational data are derived from Fig 1A.