Looking for bacterial expression plasmid SalI KpnI

[Mega [Andreas Sturm]]

Hi everyone,

Does anyone by chance have a plasmid that has some promoter, then SalI and then KpnI?

Could trade it against a plasmid or some money or thelike. 

Best,
Andreas

 

[Cathal Garvey]

If you're talking about money, it'd probably be cheaper to order it
altogether. Promoters aren't usually that large, although if you want a
*good* one you'd need a bit more length.

[Andreas Sturm]

Well, the lux operon seems to be very difficult to PCR. We tried it several times with brand new phusion (proof-reading) polymerase. No amplification.

Then we tried with phire polymerase, it works after some modification.

Phire is not proof-reading, yet it makes only half the errors as Taq. So, to see if there are no point mutations (that destroy the function of a lux gene) we'd need a promoter that reads off the lux genes in E coli. Designing new pirmers again... I'm glad these finally worked, so a plasmid with T5 or someting (not induced or induced) would seem awesome..