Manual - New England Biolabs

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Supplied Components:
The RheoSwitch ® Mammalian Inducible Expression System contains plasmids
and ligand inducer necessary to express genes in mammalian tissue culture.
Store at –20°C.
Vectors:
pNEBR-R1 Regulator Plasmid (500 µg/ml) 20 µg
Expresses an engineered nuclear receptor heterodimer, consisting of two
proteins, RheoReceptor-1 and RheoActivator. The two proteins constitute the
holoreceptor and regulate transcription of genes cloned into the expression
vector, pNEBR-X1. The two proteins are expressed from the constitutive ubiquitin
UbC and UbB promoters, respectively. The plasmid also contains the neomycin
resistance gene under control of the SV40 early promoter for the generation of
stable cell lines.
pNEBR-X1Hygro Vector (500 µg/ml) 20 µg
Used to clone the gene of interest. The plasmid, has five copies of the GAL4
response element (5XRE) upstream of a TATA box and a short leader sequence
followed by multiple cloning sites and an SV40 polyA signal. The plasmid also
contains the hygromycin resistance gene under control of the thymidine kinase
promoter for generation of stable cell lines.
pNEBR-X1GLuc Control Plasmid (500 µg/ml) 20 µg
Used as a positive control for expression analysis. This plasmid expresses
secreted Gaussia luciferase.