Team:Uppsala University/Datapage

Small RNA Scaffold

This is the native spot42 small RNA constitutively expressed by BBa_J23101 and is useful as a template for generating a randomized library of small RNAs for downregulation of any gene of interest.

Reporter System

This part can be used to insert any 5’UTR of a gene of interest to screen for downregulation of this gene by artificial small RNAs. This reporter reporter system comes with a fluorescent marker that is downregulated by any small RNA targeting the 5'UTR of the gene of interest.

New low-copy backbone

This is a new BioBrick standard vector with low copy pSC101 replication origin, especially usable for Lambda Red recombineering in E coli.

Blue Chromoprotein

aeBlue is a blue chromoprotein extracted from the basal disk of a beadlet anemone Actinia equina. The protein has an absorption maximum at 597nm and a deep blue colour. The protein aeBlue has significant sequence homologies with proteins in the GFP family.

Promotertest

We characterized this promoter in different strains as well as in one of our new lacIq backbones.

Backbones

We characterized pSB4C5 and found that it is not a low copy backbone as it's assumed to be. Plasmid prep yields, color on plate and measurements of fluorescence with flow cytometry all indicates that the copy number of this backbone is higher than our new low copy backbone BBa_K864001 and the fluorescence measurements shows that the copy number is even higher than for the medium copy pSB3C5 backbone.

eforRed

The chromoprotein eforRed naturally exhibits red color when expressed. The color is weaker than RFP, however. On agar plates and in liquid culture, the color is readily visible to naked eye in less than 24 hours of incubation. The DNA was codon-optimized for expression in E.coli and synthesized by the Korean company Bioneer Corporation.

cat-SacB

In our project we have extensively used recombineering onto the bacterial chromosome. To aid this work, we have built a improved Cat-SacB selection-counterselection cassette, which provides easy and reliable recombineering, and the possibility of scarless gene deletion.

Low copy backbone with LacIq

In addition to our new low copy backbones, we've also built variants with lacIq on the backbone which for tight repression of any lac promoter cloned into the cloning site.

Low copy temperature sensitive backbone

This backbone is a low copy backbone that have a temperature sensitive pSC101 replication origin (pSC101ts) and propagates at 30ºC but stop replicating at 42ºC.