The pSP-luc+NF Fusion Vector is a luciferase cassette vector containing the engineered firefly luciferase gene, luc+NF. The luc+NF gene is related to the luc+ gene found in the pGL3 family of eukaryotic reporter vectors but has been further modified for maximum flexibility in constructing N-terminal fusions (NF) with luciferase. Subcloning luc+NF into expression vectors provides a useful genetic reporter with exceptional sensitivity. The pSP-luc+NF Fusion Vector is not itself intended for the expression of luciferase in eukaryotic cells, because it does not contain eukaryotic promoters, enhancers or polyadenylation signals.
A unique BstEII site has been inserted immediately downstream of the luciferase ATG translation codon, allowing cloned inserts to be positioned immediately downstream of the luc+NF initiation codon. This vector is
recommended specifically for applications where N-terminal fusion proteins
do not contain an internal ATG codon at the luciferase junction.
The luc+NF gene is positioned downstream of an SP6 promoter and a ribosome binding site. An opposing T7 promoter is located immediately downstream of luc+NF. Thus, the pSP-luc+NF Fusion Vector provides a convenient template for the in vitro synthesis of both sense and antisense luciferase transcripts for studies involving in situ hybridization, RNA processing, RNA transfection or coupled in vitro transcription/translation and protein folding. Multiple cloning regions containing recognition sequences for commonly used restriction enzymes are positioned at the 5´ and 3´ ends of luc+NF to provide maximum flexibility in cloning. Luciferase enzymatic activity can be assayed most efficiently using one of the Luciferase Assay Systems.