[DIYbio] Re: Looking for pGLO

Hello
I have a similar plasmid.
map in pdf
It is located in E. coli BL21D3
If you are interested write me at E-Mal.


воскресенье, 7 апреля 2013 г., 19:49:48 UTC+3 пользователь Koeng написал:

Hey guys I want to trade some of my vectors for pGLO since I really don't wanna pay 40-50 bucks for it and have to ship it to my school. 

What I have to trade-

Vectors-

pBeloBac11

pUC19

Carolina's pGreen

pVIB

Strains-

Halobacterium NRC-1

I also have a Halobacterium vector I am testing right now. It is biobrick compatible, chloramphenicol resistant, shuttle vector with E coli. I am still testing it, (gonna make a new batch today cause I found out that mutagenesis products are linear...) and it should be about a week before I have a bunch of it. Here is the plasmid it is "based" off of.

http://www.atcc.org/Products/All/77216.aspx

I pretty much used a viral origin of replication for the Halobacteria, pUC origin for E coli, and chloramphenicol resistance from pBeloBac11. I also added a synthetic RBS before the chloramphenicol gene so (hopefully) it can be expressed in halo bacteria. However, I kept the same promoter because I think that the sigma factor for E coli may also work a little in halobacteria, added that I put a RBS site to maximize production. If ya wanna try the vector, just get some halobacteria broth and agar from carolina - http://www.carolina.com/prepared-biological-media/halobacterium-sp-nrc-1-media/FAM_775950.pr?catId=&mCat=&sCat=&ssCat=&question=halobacteria+media

I will give ya some of the strain for free :)

-Koeng

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