pCMV-CLuc 2 Control Plasmid
The pCMV-CLuc 2 Control Plasmid is a mammalian expression plasmid that constitutively expresses the secreted luciferase from the Ostrapod Cypridina noctiluca (CLuc) under the control of the CMV promoter.
- CLuc can be assayed in the culture medium of mammalian cells expressing Cypridina
- CLuc does not use the same substrate as other marine luciferases, e.g., Renilla and Gaussia. Therefore, it is possible to assay both CLuc and GLuc independently in cell culture medium from cells expressing both reporters.
- Can be used to generate Neomycin-resistant stable cell lines
The pCMV-CLuc Control Plasmid is a mammalian expression vector that encodes the secreted luciferase from the Ostracod Cypridina noctiluca as a reporter, under the control of the constitutive CMV (cytomegalovirus) promoter. Cypridina luciferase (CLuc) is a 62 kDa protein with a native signal peptide at the N-terminus that allows it to be secreted from mammalian cells (1). Because it is secreted CLuc can be detected in the culture medium of mammalian cells expressing the reporter gene. 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.
Recommended sequencing primers for pCMV-CLuc 2 Control Plasmid (not available from NEB)
T7 Universal Primer (20-mer)
pBasic Reverse Primer (25-mer)
CLuc 3´ End Forward Primer (23-mer)
CLuc 5´ End Reverse Primer (24-mer)
- CMV promoter: 209-863
- CLuc coding: 919-2580
- Start codon: 919-921
- Stop codon: 2578-2580
- Signal peptide: 919-972
- Synthetic poly-A site: 2589-2637
- Neo promoter (SV 40): 3223-3558
- Neomycin resistance gene: 3623-4404
- Bacterial replication ori (pMB1): 5738-
- Amp resistance: 6769-5909
- All pLuc-2 vectors have improved polyadenylation-transcription termination of the luciferase transcript. The polyadenylation signal is a synthetic polyadenylation sequence based on the β-globin gene (4).
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