Hello everyone. I'm a student working on a research project which also has some plasmid design involved. This isn't my strong point. I know I'm a noob so I understand if you laugh at my lack of knowledge.

The idea is as follows: Working with the HEK293 cell line. I need to swap out a regular house-keeping gene, and swap-in the same gene with a GFP alongside it. So i thought I would choose Beta Actin (https://www.ncbi.nlm.nih.gov/gene/60). Basically the swap in gene would be a construct of Beta Actin + GFP (is this doable?)

We are outsourcing the CRISPR design to a company, but they require us to give them the donor template which we want swapped in.

Would the following construct work? - 200bp homology for target site (I'm not sure how I find this) - Beta Actin Gene - GFP gene - 200bp homology for target site (again, not sure how to find this)

Here is a screenshot of what I have so far in Gene Compiler: Any help is appreciated!