Hi everyone,
Back again on this forum first time since 2017!
I don't know if anyone has seen Justin Atkins' video on his YouTube channel
https://www.youtube.com/watch?v=20vwKHfbVvY&t=7423s,
involving using a subunit of a malarial protein that seems to bind specifically to placental and cancer cells
Well, it caught my eye, so I tried following the steps in his video (using it as a tutorial) using Benchling and Addgene for the plasmid:
URL: https://www.addgene.org/browse/sequence/354458/
Unfortunately, the restriction sites that he suggested using in the fusion protein he created in silico also occur elsewhere in the plasmid that I want to insert the sequence into, so in my version of the fusion protein, I removed all the cloning sites, and attached two dummy CRISPR sequences at the beginning and end of the sequence
Here's the protein I've "made":
https://benchling.com/scratch/f/lib_7sFnbNIl-membrane-binding-sequence-var2csa-codon-optimised/seq_rT20ZjYa-var2csa-binding-subunit-human-fc-fusion-protein-with-ig-secrtion-tag/edit
My question is: can you just add a target sequences willy-nilly to an already existing sequence and insert it into any plasmid, (such as the one I am using) or does it need to be a special kind of plasmid?
Advice will be appreciated!
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