pNL3.2.CMV Vector

The pNL3.2.CMV Vector constitutively expresses the NanoLuc reporter fused to a PEST destabilization domain (NlucP). The NlucP fusion protein naturally accumulates at low intracellular levels due to constitutive proteosomal degradation, thus serving as a negative control for experiments configured to measure regulated changes in NanoLuc luciferase expression levels (e.g., NanoLuc fusion vectors). This vector also encodes a hygromycin resistance gene for selection in mammalian cells. NanoLuc (Nluc) luciferase is a small enzyme (19.1kDa) engineered for optimal performance as a luminescent reporter. The enzyme is about 100-fold brighter than either firefly (Photinus pyralis) or Renilla reniformis luciferase using a novel substrate, furimazine, to produce high intensity, glow-type luminescence. The luminescent reaction is ATP-independent and designed to suppress background luminescence for maximal assay sensitivity.