When constructing our BioBricks, we made use of the IDT synthesis offer. This way we were able to generate our required parts in a fast and efficient way. Additionally, we made use of PCR amplification with our constructed expression plasmids as template. All of the parts contain a coding sequence, some of which extended with promoters or introns. Since our fusion protein BBa_K2643000 consists of the dxCas9, linker and Tn5 coding sequences this part is registered as composite part. The basic parts comprising the our fusion protein composite part are documented separately and also exist as separate BioBricks. This is advantageous for those wishing to combine one of our basic parts with a different expression level or compose a different composite part altogether. These basic parts were generated by using PCR on the composite part and restriction ligation cloning. We created a part collection comprised of 6 basic parts encoding for different EPO coding sequences, 3 basic parts encoding for the required components for targeted sequencing and 1 composite part of our fusion protein. All BioBricks are submitted with elaborate characterisation so the user may choose the best protein for their project accordingly. For the composite parts of the TDPs we designed a new, strong RBS. This part does not exist separately. Additionally, we include the documentation of a basic part encoding the gene for Cas13a, derived from the plasmid pC011 from the authors of Gootenberg et al. 2017. This BioBrick does not exist separately. Finally, we submitted a composite part containing a nonsense spacer with flanking double repeats (with constitutive promoter), appropriate for use for Cas13a. This part is designed so that the user can easily change the spacer according to the sequence they wish to target. Find more info on this cool part on its page in the registry! We also documented this as a basic part without a promoter. This part does not exist separately. Our favourite basic part is BBa_K2306003 and our favourite composite part is BBa_K2306008.

Parts Overview