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cDNAs encoding full-length CaMKII variants were subcloned into a pET vector containing N-terminal 6xHis followed by a SUMO tag. CaMKII chimeras were constructed as follows. The cDNAs encoding the kinase/regulatory segment and hub domains of CaMKIIα and CaMKIIβ were amplified from full-length constructs. The kinase domain includes the regulatory segment of its corresponding kinase. Each domain was amplified with a 20-bp overhang that was the corresponding CaMKII gene. The chimeras were assembled using Gibson assembly.
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