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His-tag fusion protein in pET-28a(+)
I am student only getting familiar with gene cloning.
I want to insert a gene in this vector (pET-28a(+), in order to get a his-tag. My question is: do you have to worry about the stop codon of the insert itself, because the vector already has a stop codon after the His-tag.
start--Histag---(start - Coding sequence -stop)----Histag--stop.
So, there are two stop codons, how do i know that translation does not stop before the his-tag is completely incorporated?
Re: His-tag fusion protein in pET-28a(+)
Those are not the same His tag. There is an N terminal, and optional C terminal tag if you do not incorporate a stop codon.
|fusion , fusionprotein , his-tag , histag , pet28a , protein , stop codon , vector|
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