IRESite record type: plasmid_with_promoter_and_putative_IRES_without_translational_characterization
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 in reverse
complement orientation 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), cloned in reverse complement orientation, and
firefly luciferase coding region (luc version of the luciferase gene) without 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): 313-1968
Remarks:
IRESite notes about verification of plasmid sequence:
The complete sequence of the BiP 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 plasmid was used as the negative control for experiments in article Enhancement of IRES-Mediated
Translation of the c-myc and BiP mRNAs by the Poly(A) Tail Is Independent of Intact eIF4G and PABP (Thoma et
al., 2004; PMID: 15383282).