I made multiple recombinant Lrp6 with different tags (I used the Gateway system, using a p221Lrp6 and a gateway based expression pCS2 vector). I could get the plasmid with Lrp6-GFP and Lrp6-Flag, only when I used the Stbl3 at 30 C. However I need another tag and I cannot get the plasmid using the same conditions. Upon sequencing, I realized that the Lrp6 gene is being excised out (most of it, leaving only a fragment of 1kb).

Do you think growing the bacteria at a lower T (maybe 25 C) could help?

What else could I do?

Thanks!