Need yeast shuttle vector

[Marc Juul]

I am part of the Counter Culture Labs crew in Oakland, California, and we're looking for a yeast shuttle vector, preferably with some or all of the following features:

  * URA3 gene
  * Ampicillin resistance gene
  * 2-micro ori

  * An E. coli ori like pUC, pBR322 or f1

  * Alpha factor secretion signal

We realized that we can't use addgene.org since their materials transfer agreement would put limits on our use and sharing of constructs.

Anyone have an IP and MTA unencumbered shuttle vector we can have?

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marc/juul

[Josiah Zayner]

I would just email the lab directly. If you pay the shipping I can't imagine a reasonable lab not sending you a vector. This is usually the way it is done in professional circles.

Also, since most of the plasmids on addgene have been published on they are public domain anyway. I always wonder if their MTA is even possible to uphold?

#1 How could anyone possibly determine if the plasmid you used is from addgene?
#2 If the plasmids are published on and public domain, doesn't that negate the MTA regardless?

I don't know much about this legal stuff so...

I think addgene are huge dicks because they sell plasmids for ridiculous amounts after they acquire it for free from a lab.

[shamrock]

Hi Marc,

I've got YEp-24 available. It has everything your looking for except for the alpha factor secretion signal. I can send you an E. coli culture with the plasmid next week if that will work for you.

Tom

BUGSS

[Nathan McCorkle]

the NEB protein expression kit has a plasmid like this... pKLAC2

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-Nathan

[Mega [Andreas Stuermer]]

Yeah, but pKlac is an integrative vector... But it's awesome. Some day I'll have enhanced thermostable GFP under a very strong promoter subcloned into it and bake real fluorescent bread. Atm no time unfortunately. And synthesis isn't cheap yet either. Would surely burn 200€ alltogether.

[EdwardPerello]

I have a custom Ecoli vector with 2 micron, HI33, AmpR and an F1 origin (its built on pgem-3z, PRS403 and YEP21). Shamrock might be able to provide you with what you need more easily however.  

[EdwardPerello]

Well Josiah, I dont think it could be determined if you used the plasmid directly from addgene, they don't barcode anything in that respect. They  could conceivably sequence your plasmid vs their plasmid too check, but they rarely sequence verify their plasmids the whole way through (usually only part of the plasmid is sequenced). The MTA is done more as a record of materials transfer as opposed to licensing the material to be used by the recipient. 

I don't think Addgene are dicks - they are a non-profit organisation and need to keep their good work going!

[Marc Juul]

On Mon, Jun 2, 2014 at 9:53 AM, shamrock <thmsb...@gmail.com> wrote:

Hi Marc,

I've got YEp-24 available. It has everything your looking for except for the alpha factor secretion signal. I can send you an E. coli culture with the plasmid next week if that will work for you.

Thanks a lot shamrock!

And thanks to everyone else who wanted to help!

YEp-24 totally fills our needs and we've coordinated off-list.

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marc/juul