pSV40-CLuc Control Plasmid
The pSV40-CLuc Control Plasmid is a mammalian expression plasmid that constitutively expresses the secreted Cypridina luciferase (CLuc) under the SV40 early promoter.
- Moderate constitutive expression of CLuc
- 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.
- CLuc can be assayed in the culture media or cell lysate
The pSV40-CLuc Control Plasmid is a mammalian expression vector that encodes the secreted luciferase from the Ostracod Cypridina noctiluca (1) as a reporter, under the control of the constitutive SV40 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) so that CLuc activity can be detected in the culture medium. There is a multiple cloning site (MCS) upstream of the SV40 promoter.
Recommended sequencing primers for pSV40-CLuc:
Forward primer (23-mer) (not available from NEB)
Reverse primer (24-mer) (not available from NEB)
- Polylinker MCS: 1–51
- SV40 promoter: 51-246
- CLuc ORF: 291–1952
- Start codon of CLuc: 291–293 Stop codon: 1950–1952
- Signal peptide: 291–344
- SV40 poly-A site: 1967–2188
- SV40 enhancer: 2195–2441
- Bacterial replication ori (pMB1): 3347–2759
- Amp resistance: 4378–3518
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