generating an empty vector!

Sudheendra Rao N R via methods%40net.bio.net (by sudhee26 from gmail.com)
Tue Jul 15 14:14:07 EST 2008


Hi DK,
I agree. common Kozak being (G/A)NNATGG
Whats that "putative transcriptional start" after 3' end of CMV promoter?
http://tools.invitrogen.com/content/sfs/vectors/pcdna3.1v5histopo_mcs.pdf
TAG is amber codon..i dont get this!!

Sudheendra.

On Tue, Jul 15, 2008 at 7:04 PM, DK <dk from no.email.thankstospam.net> wrote:

> In article <mailman.745.1216125714.3533.methods from net.bio.net>, "Sudheendra
> Rao N R" <sudhee26 from gmail.com> wrote:
> >Hello,
> >as far as remember, pcDNA3.1 does have a putative transcription
> start..wont
> >that make sure that tags are expressed even in empty vector which has got
> >self ligated..
>
> No, it won't. The plasmid lacks *translation* start (that needs to be
> supplied with a cloned gene).
>
> http://tools.invitrogen.com/content/sfs/vectors/pcdna3.1v5histopo_mcs.pdf
>
> Off topic note:
> Which means that if only ORF, without any extra sequences is
> cloned, the immediate sequence upstream of the AUG will be
> GCCCTT - don't think it it fits Kozak consensus very well. So,
> likely relatively poor expression levels can be expected.
>
> DK
>
>
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