Experimental localization. i) Rapid PCR generation of tagged expression constructs. A linear expression construct is produced by fusing three fragments: a CMV promoter, a dual SV40 terminator and the myc tagged open reading frame to be tested. The epitope tag is added by use of a gene specific primer that fuses the epitope in-frame with the open reading frame. Overlap sequences corresponding to AL41/GSN1 and P15/AL25 are used to prime off each other to generate a full length construct consisting of all three products. Nested primers AL35 and AL38 are used to further amplify the construct. Primer sequences and C-terminal schematics available as Supplemental data. ii) Representative observed localizations. A) Cytoplasmic: (left to right) Rps6kb1, 4932415A06Rik, Camk2d, Lats2 B) Ubiquitous, nuclear and cytoplasmic: Csnk1a1, Camkk2, Stk35, Mapkapk3 C) Nuclear: Dusp4, Mastl, Smok1, Trib1 D) Membrane associated: Ptpn5, Txk, Ptp4a3, Vrk2. Bar 10 μm.