Dear Dicty community,

We are planing to knock in a gfp tag downstream a gene of interest. Therefore the upstream flank of the construct lies in the coding region of the gene. To generate the knock-in construct we had to introduce a restriction site at the 5'end of the upstream flank.
Our question is, if the few additional basepairs of the restriction site will also be integrated into the genome (even though, they are not part of the homologous region). In our case this would lead to a frameshift within our target gene which would be detrimental.
Are there easy ways to get around this problem (if no endogenous restriction site is present)?
  
Greetings from Hamburg,
Frauke


Bernhard-Nocht-Institute for Tropical Medicine
Research Group Hagedorn
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