Team:TU Munich/project/data

Data For Our Favorite New Parts

BBa_K568003 - T7 promoter with lacZ reporter part:

Our original reporter construct. The presence of a functional T7 polymerase causes expression of beta-galactosidase. Thus, it is possible to detect the output of our optogenetical AND-Gate.

BBa_K568002 - blue light promoter with lacZ reporter part:

The blue light promoter followed by lacZ is a simple optogenetic device that expresses beta-galactosidase if the blue light input is positive. We used this part to find out the characteristics of the response to blue light in E. coli. The blue light promoter is part of the YcgE/F system which is a blue light sensor that is naturally found in E. coli. Thus, only the promoter itself is needed in this part.

BBa_K568001 - optogenetical AND-Gate red/blue light:

This part is the centerpiece of our 3D-printer. It is supposed to cause expression of a functional T7 polymerase only when both the red light and the blue light input are positive. Further characterization will be needed.

All submitted parts

BBa_K568000 - red light sensor:

A red light sensor without reporter. It was extracted from BBa_K322127 using PCR. Thus this is an improvement of BBa_K322127 by making it usable in many different contexts.

BBa_K568001 - optogenetical AND-Gate red/blue light:

see above ("favorite parts") for information concerning this part.

BBa_K568002 - blue light promoter with lacZ reporter part:

see above ("favorite parts") for information concerning this part.

BBa_K568003 - T7 promoter with lacZ reporter part:

see above ("favorite parts") for information concerning this part.

BBa_K568004 - optogenetical AND-Gate without first promoter - choose your first input:

This part is a part of the optogenetical AND-Gate. It lets you choose the first input, that in combination with blue light will lead to expression of the T7 polymerase. Whether you choose a different wavelength of light, a chemical substance or a physical condition - the choice of the first input is up to you!

BBa_K568005 - Intermediate optogenetical AND-Gate without T7ptag:

This part is a precursor of the finished optogenetical AND-Gate. You could add any coding sequence carrying one or multiple amber mutations behind this part which would result into expression of the desired protein when irradiated by both blue and red light.

BBa_K568006 - Intermediate synthetic part:

This part is contained in the optogenetical AND-gate. This part consists of 5 subparts which are very small. Because cloning such small parts is difficult and time consuming, we decided to have this part synthesized by GeneArt.

BBa_K568007 - GFP under constitutive promoter:

This part serves as a positive control in our quantitative GFP assays. It consists of gfp with rbs and terminator, which is under control of a constitutive promotor.

BBa_K568008 - Red light promotor with GFP reporter:

This part was constructed so that we were able to characterize our red light promotor with a quantitative GFP assay in addition to our lacZ Miller assay. It consists of gfp with rbs and terminator, which is under control of the red light promotor.

Data For Pre-existing Parts

Experience - BBa_K322127 (Edinburgh, 2010): Phycocyanobilin synthesis genes and red light detection system
We used this part in two ways: First of all, we used PCR to amplify the parts needed for red light detection from this part, generating BBa_K568000. Secondly, we compared parts BBa_K322127 and BBa_K568000 using a Miller Assay. Unfortunately, this part did not produce the results we hoped for.

Experience - BBa_I746907 (Cambridge, 2007): T7 promoter driving 6-his tagged P7 GFP

We used this as additional reporter plasmid and did an IPTG induced GFP assay to verify the part. It worked perfectly for us.