• All pGLuc 2 vectors and plasmids have improvedpolyadenylation-transcription termination of theluciferase transcript. The polyadenylation signalis a synthetic polyadenylation sequence based onthe â-globin gene (5)
• 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
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 12 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.
Recommended Sequencing Primers for pGLuc-Basic 2 Vector (not available from NEB)
Upstream of MCS: (23-mer) 5´-GGGGTTCCGCGCACATTTCCCCG-3´(4917–4939)
pBasic Reverse Primer (25-mer) 5´-TCAGAAGCCATAGAGCCCACCGCAT-3´(785–761)
GLuc 3´ End Forward Primer (20-mer) 5´-GCCAGCAAGATCCAGGGCCA-3´(580–599)
GLuc 5´ End Reverse Primer (24-mer) 5´-TCAGGGCAAACAGAACTTTGACTC-3´(103–80)
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