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Cloning steps for each heavy and light chain are in Geneious in the "New antibodies folder" under "BCR 2.0".  Amino acid sequences were back-translated using IDT and , optimizing for gBlocks (there were no BsaI cut sites were manually eliminated.  Since to eliminate).

Since we had issues with mutations when we ordered the gantenerumab heavy chain as one gBlock and because ordering the entire heavy chain costs ~$about $240 more (per sample) than just ordering the variable region, I propose PCRing out the mIgM constant region from Gmab heavy and Golden Gating the variable and constant regions together.  This way we can also design a vector that makes variable region swapping easier in the future (since we're only testing a couple of antibodies though, I think that making that vector can wait until later, vs. the linkers where we are testing so many of them).  Primers for that PCR are in the "IgM heavy chain primers" folder.  The primer regions don't make it look like we'll have too many problems...

VRC01 and HyHEL-10 gBlocks are located in their respective folders, along with the pENTR that they will make when ligated into the Golden Gate backbone.

BUDGET:

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