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: 3UTR_incomplete
The mRNA/+RNA description:
Putative bicistronic mRNA transcript derived from pBSRLucEFluc plasmid through SV40 promoter. The plasmid has
in the intercistronic region T7 promoter.
The mRNA/+RNA sequence represented in the +DNA notation:
Credibility of mRNA sequence: reverse_engineered_sequence_and_should_match_experiment_maybe_except_3UTR
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): 2
Notable Open-Reading Frames (ORFs; protein coding regions) in the mRNA/+RNA sequence:
The description of the protein encoded in this ORF: Renilla 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): 2636-3571
Remarks:
Plasmid DNA was digested by EcoRV/SalI and BamHI/SalI giving fragments 4321/1965 bp and 3392/1877(17) bp,
respectively. Amplification with Firefly/Renilla gene specific primers (Fluc_seq, gTg gAC gAA gTA CCg AAA gg,
position within plasmid sequence:2118, Rluc_revseq, CTGCATGTTTTTCTGAATC, position within plasmid
sequence:3060) gave one fragment of expected size. The sequence of promoter and intercistronic regions was
determined by sequencing (regions 54-626 and 2180-2958 of plasmid sequence). More detailed data are annotated
in GenBank sequence file.