Mammalian expression vectors with HA, c-myc, FLAG or anyother tag - The lab is running out of funds and need some cheap source. (Jun/12/2009 )

Hi All,
It will be great if somebody can tell me the source for mammalian expression plasmids with tags.
Looking forward to hear from you.
Regards,
Niraj

nirajm on Jun 12 2009, 08:10 PM said:

Try Addgene (http://www.addgene.org/pgvec1). It is a share site that provides plasmids submitted from academic groups in the same way that you request vectors from other labs. You can obtain numerous vectors for about $65 ea. if you sign an MTA.

There is also http://plasmid.med.harvard.edu/PLASMID/Ord...erview.jsp#cost for about $45/clone.

Also if you have an empty vector and you want to add a tag, just add the tag to a PCR primer for a target gene and amplify from there. If you add a restriction site on either side of the tag, you should be able to chop it out and re-liagte it into other plasmids too.

bob1 on Jul 22 2009, 07:33 PM said:

I just did this myself using FLAG. Added it to pBluescript, and then added a restriction site in front of the tag, so I can make C-terminal FLAG constructs. Can also insert flanking sequence downstream of the tag if I ever want to express the tagged protein as a single copy in the genome as opposed to plasmid-borne. Went together pretty easily. It required some polymerase, restriction enzymes, and some sequencing. All in all, likely cheaper than what I would've paid (was looking at Sigma for FLAG vectors), and probably more versatile with being able to add flanking sequence behind the tag.


As for places that sell vectors, I remember seeing many at Sigma for FLAG. Almost any supplier of molecular biology supplies probably has a good number of tag vectors to choose from. But as stated above, you'll probably pay at least 5 or 6 times more than if you'd get one from the links above.