BioBricks overview

Below, you can find all the biobricks we created!

Besides, we submitted one part that cannot be classified as a biobrick because it has some illegal restriction sites and it is in the pSB1A3 backbone:

Initally, part BBa_K1913015 was made only to for testing the in vitro assay, jacco, please put the rest of your excuse here ;).

No Cas9 biobrick?!
When we started making the constructs for the Cas9 kill switch, two approaches were taken: one was taking the Cas9 that is available in the iGEM registry (BBa_K1218011) as a starting point for making mutations and expressing Cas9. The other approach was starting with pdCas9 (Addgene plasmid # 46569). While cloning, it proved to be difficult to transfer BBa_K1218011 to another backbone (see also the notebook). Furthermore, when cultures transformed with BBa_K1218011 were checked for Cas9 expression with SDS-PAGE, no convincing Cas9 band could be observed. Because of time limitations we decided to continue working with the Addgene construct for making the mutations, and chose an established system for protein expression. For that reason, no Cas9-biobricks were submitted. Furthermore, the pEVOL construct containing an aminoacyl-synthetase and a tRNA for introducing BipA in response to the TAG stopcodon were isolated from a strain kindly received from George Church (described in Mandel et al., 2015). The MTA that was signed to receive the strain does not allow for redistribution.