Sequences Needed, mYFP, mCFP, tdTomato - We are missing these sequences :(( (Feb/15/2010 )

Hi everyone,

We've recently obtained these 3 fluorescent protein expression plasmids from Dr.Roger Tsien's lab.

Unfortunately they did arrive with no documents. Their online lab website does have limited information. I've also tried to contact 2-3 times with email, got no response.

All I know is tdtomato is in pREST-B vector, but don't have the whole sequence, other two I am just guessing they are in pCDNA3 or pCDNA3.1 vector, but again don't have conclusive evidince...

If you could give me some insight, I would really appreciate that!

Do you want the sequence of protein or vector?

The sequence of these fluorescent proteins should be available. google it. Just PCR amplify it and clone into vector of choice.

scolix on Feb 15 2010, 09:44 AM said:

I need vector information...

sequence the vector, and you should have the vector sequence and MCS

scolix on Feb 15 2010, 10:53 AM said:

Thanks, I thought that for sure. But I really don't want to sequence whole vector, these seems to me quite common vectors, it is surprising nobody has any info about them.

Try with NCBI looking for sequences of vectors in "nucleotide".
For example, here tere is the pcDNA3.1+PA sequence: http://www.ncbi.nlm.nih.gov/nuccore/EF5502...quence_RVDocSum

I hope I have been of some help.

Ilaria

Hey,

for pcDNA3.1. just check invitrogen.com (for example: http://products.invitrogen.com/ivgn/product/V79020) and you will find everything. Just sequence out of CMV promotor and you have the CMS and (part of) the insert.

Never heard of the other vector before. But theres a pDEST-B vector on invitrogens page. Maybe it is this one?

Greetings,

Franzi!

KnightFall on Feb 15 2010, 07:37 PM said:

people usually PCR amplify the gene and clone into vector of choice. Dont bother too much with the vector it is in.