Does the presence of an attR1 sequence (~ 120bp) in between a 3xFlag sequence and the N-terminal of gene of interest affect translation of the gene.
The backbone destination vector has the 3X Flag sequence followed by attR1 sequence. After recombination, the attL flanked gene from entry clone is going to replace the region downstream of attR1 in destination vector.
I was wondering if ~120bp (of attR1) in between the 3x FLAG sequence and gene of interest will affect translation?
Thanks for your input.
After the LR recombination reaction, you should be left with a 25 bp attB sequence between the 3xFlag sequence and the gene coding sequence. As long as your gene coding sequence is in frame with the 3xFlag tag, this should translate with a short peptide linker in between.
https://tools.thermofisher.com/content/sfs/manuals/gatewayman.pdf
@ Alex. Makes sense, thanks a lot for the input and attachment.
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