DNA plasmids and cell lines

Attached are the sequences of RA-Sec61B and GB-C1 (there was no GFP encoded here) plasmids. 

RA-Sec61B - The sequence regions encoding for RA tag and Sec61B are 597-1289bp and 1311-1601bp, respecitively. 

1. The RA tag was PCR amplified from Addgene plasmid #61019 with overhangs encoding for NheI and BspEI cut sites.
2. The Clontech vector plasmid pAcGFP1-C1 was sequentially digested using NheI and BspEI restriction enzymes to remove GFP encoding region. 
3. RA tag was ligated into the digested pAcGFP-C1 vector to create RA-C1 vector.
4. Sec61B was PCR amplifed from Addgene plasmid #49155 with overhangs encoding for XhoI and KpnI cut sites.

5. Sec61B was ligated into XhoI and KpnI digested RA-C1 to create RA-Sec61B.

    

GB-C1 empty vector - The sequence regions encoding for GB tag are 597-1274bp with the multiple cloning site encoded by 1275-1446bp.

1. The GB tag was PCR amplified from Addgene plasmid #61017 with overhangs encoding for NheI and BspEI cut sites.
2. The Clontech vector plasmid pAcGFP1-C1 was sequentially digested using NheI and BspEI restriction enzymes to remove GFP encoding region. 
3. GB tag was ligated into the digested pAcGFP-C1 vector to create GB-C1 vector.

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