Perform a double digest of XhoI and NdeI on pET29b+ and purify the resultant 4.5 kb fragment (the backbone) using a gel purification kit. Dilute the prepared backbone to 40 ng/µL for downstream applications.
Order your codon-optimized synthetic gene sequence from your vendor of choice (we use Life Technologies in the Siegel group) with the following sequences tacked on to the ends (these are specific to pET29b(+))