pGLuc Mini-TK 2 is a cloning vector for mammalian cells, containing a minimal promoter fragment from the HSV thymidine kinase (TK) promoter adjacent to a reporter gene, 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 Mini-TK 2 Vector contains a multiple cloning site (MCS) upstream of the minimal TK promoter for cloning promoter or enhancer elements. A neomycin resistance gene under the control of the SV40 promoter allows selection for stable integration of the plasmid into the mammalian cell genome using G418.
- Polylinker MCS: 12–68
- Minimal promoter from Herpes simplex virus Thymidine Kinase (Mini-TK): 69–131
- GLuc coding: 146–703
- Start codon: 146–148
- Stop codon: 701–703 • Signal peptide: 146–196 • Synthetic poly-A site: 712–760
- Neo promoter (SV40): 1346-1681
- Neomycin resistance gene: 1733-2527
- Bacterial replication ori (pMB1): 3861-3273
- Amp resistance: 4892-4032
- All pGLuc-2 vectors have improved polyadenylation- transcription termination of the luciferase transcript. The polyadenylation signal is a synthetic polyadenylation sequence based on the b-globin gene (5).
Product SourceIsolated from E. coli strain NEB 10-beta by a standard DNA purification procedure.
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