Table 3 DNA Oligomers used in the molecular cloning of RABEP2.

From: Rab-GTPase binding effector protein 2 (RABEP2) is a primed substrate for Glycogen Synthase kinase-3 (GSK3)

A) PCR primers

Sequence

pGEX fwd

GAATTC GACTACAAGGACGACGATGACGCGGCAGCTGCGCCGGTGG

pCMV fwd

GAATTC GCCACCATGGACTACAAGGACGACGATGACGCGGCAGCTGCGCCGGTGG

rev

GAATTCTCAGGTGTCCTTGATGTCCCTGACG

B) Mutagenesis

Sequence

S200A fwd

GTTGCTGCCCCTGGCCCGGGATCCATC

S200A rev

GATGGATCCCGGGCCAGGGGCAGCAAC

S204A fwd

CTGTCCCGGGATCCAGCGCCCCCG

S204A rev

CGGGGGCGCTGGATCCCGGGACAG

S204E fwd

CCCTGTCCCGGGATCCAGAGCCCCCGCT

S204E rev

AGCGGGGGCTCTGGATCCCGGGACAGGG

  1. (A) Primer sequences used to generate hRABEP2 cDNA for subcloning into pGEX and pCMV expression vectors. EcoR1 restriction sites are underlined, a Kozac sequence is in italics, while the FLAG tag sequence is in bold. (B) Site directed mutagenesis primer sequences with the base changes introduced to produce amino acid modifications shown as underlined and in bold.
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