That's what I meant. I need an mcs with nco flanked by eco and sac.. Nco is a great stand-in for a kozak sequence in plants (ccatATGg rbs) and I have primers for adapting nco to the five prime of many genes of interest but at the moment I lack a plasmid that has nco where I need it. My plan is to have a modular system for quick transient expression tests using eco-nco as a promoter region, nco sac as the gene of interest region and sac kpn as the terminator. Already tested this in silico and its compatible with a number of go-to plant parts so all that's left is a good mcs. Again, ideally EcoRI-NcoI-SacI. Thanks again for the insight.
Sebastian S. Cocioba
CEO & Founder
New York Botanics, LLC
Plant Biotech R&D
Sebastian S. Cocioba
CEO & Founder
New York Botanics, LLC
Plant Biotech R&D
From: Josiah Zayner
Sent: 10/29/2014 7:03 PM
To: diybio@googlegroups.com
Subject: [DIYbio] Re: Plasmid request
This might be hard to find because NcoI is CATATG and is usually used at start codons so usually one of the first in the MCS. After looking at a few vectors it seems to be the case but if you could perhaps find a vector that has EcoRI first and put out a request for that maybe myself or someone else could provide the specific vector.
Josiah Zayner, Ph.D.
NPP Fellow
NASA Ames Research Center
Space Biosciences
On Tuesday, October 28, 2014 11:22:26 AM UTC-7, Sebastian wrote:
-- Josiah Zayner, Ph.D.
NPP Fellow
NASA Ames Research Center
Space Biosciences
On Tuesday, October 28, 2014 11:22:26 AM UTC-7, Sebastian wrote:
I'm looking for a bacterial plasmid with an mcs containing EcoRI, NcoI, and SacI in that order. I doesn't need to be a blank shuttle, just any working plasmid with those sites and amp resistance preferably. Need to shuttle in the NcoI site and don't want to design primers, use hf polymerase, etc etc. I'll pay for shipping and fee if needed. Thanks a bunch!
Sebastian S. Cocioba
CEO & Founder
New York Botanics, LLC
Plant Biotech R&D
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