• 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
Description:
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:
CLuc:
Forward primer (23-mer) (not available from NEB)
5´-GAGTTCAAGAAAGAATGCTACAT-3´ (1888–1910)
Reverse primer (24-mer) (not available from NEB)
5´-GTAAGGACAGTCCTGGCAATGAAC-3´ (360–337)
Source:
Isolated from an E. coli strain NEB10β by standard DNA purification methods.
Applications:
The pSV40-CLuc Control Plasmid can be used as a control for assessing the efficiency of transfection in mammalian cells. Plasmids containing other constitutive promoter elements are also available (see Companion Products Sold Separately). CLuc can be used as a stand alone reporter or in conjunction with other compatible reporters such as Gaussia luciferase (GLuc)(2). CLuc and GLuc are ideally suited for co-expression as both are secreted and highly active enzymes providing ease of use and sensitivity.
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