I have a problem with expression of a protein which has second residue exactly what I want to express.
I need to exrpress Mat-Lys~ and Mat-Asn with pAcycDuet vector which has His tag in MSC.
Because I want to delete His tag, I have no choice except Nco1 for RE.
Also. it is very important to express Lys or Asn as a seond residue in my protein.
Here is a problem!
Nco1 has ATG start codon in the restriction site and is followed G.
I mean Nco1 site is CCATGG, so I can not express Lys( AAA) or Asn(AAT) as a second residue after Mat because of G after ATG in Ncoi site.
What should I do? plz help me.
Have a good day~^^