Description:

pGLuc-Basic 2 is a cloning vector for expression in mammalian cells, containing a reporter gene but lacking promoter elements. The reporter gene is the secreted luciferase from the copepod Gaussia princeps. Gaussia Luciferase (GLuc) is a 19 kDa protein encoded by a \"humanized\" sequence, and it contains a native signal peptide at the N-terminus that allows it to be secreted from mammalian cells into the cell culture medium (1,2). The pGLuc-Basic 2 Vector contains a multiple cloning site (MCS) upstream of the GLuc coding sequence. A neomycin resistance gene under the control of an SV40 promoter allows selection for stable integration of the plasmid into the mammalian cell genome using G418.Figure 1: Comparison of light output obtained from HEK293 cells transfected with either the promoterless pGLuc-Basic 2 Vector or the pCMV-GLuc 2 Control Plasmid (NEB #N8081). Supernatants were harvested at 24 hour post-transfection and assayed for the GLuc activity using the BioLux GLuc Assay System.Figure 2: pGLuc-Basic multiple cloning site (MCS)The Gaussia Luciferase sequence is shown with a blue background. Only unique restriction sites are shown.

DNASU and Addgene are central repositories for plasmid clones and collections that may also be helpful.

Highlights

• Polylinker MCS: 12–68
• GLuc coding: 76–633
• Start codon: 76–78
• Stop codon: 631–633
• Signal peptide: 76–126
• Synthetic poly-A site: 642–690
• Neo promoter (SV40): 1276–1611
• Neomycin resistance gene: 1663–2457
• Bacterial replication ori (pMB1): 3791–3203
• Amp resistance: 4822–3962
• All pGLuc 2 vectors and plasmids have improved polyadenylation-transcription termination of the luciferase transcript. The polyadenylation signal is a synthetic polyadenylation sequence based on the β-globin gene (5)

Product Source

Isolated from an E. coli strain NEB10b by standard DNA purification procedure.