Your target gene can be inserted via the Bam HI- site and one of the restriction sites downstream of it.
This Ser will be left in your protein after cleavage with SenP2. In any case, you have to make sure that you have the terminal two Glycine codons of SUMO3 in-frame with your target gene. Note : The first amino acid after the Glycines must not be Proline!
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The protease cannot cleave the protein in that case! You can also obtain your target protein with a C-terminal His 6 -tag if you don't add a stop codon to your gene. In this case, please make sure that the second His6-tag is in-frame with your gene.
Proteome-wide identification of SUMO modification sites by mass spectrometry
Depending on the 3'-restriction site, you may have to add one additional base. Part I. Ulrich Part II.
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