Plasmid project

41 views

Skip to first unread message

Simon Rose

unread,

Jul 1, 2025, 10:28:44 AMJul 1

to DIYbio

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?