Team:TUDelft/Parts/Main
Parts Overview
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, 2 basic parts encoding the required components for targeted sequencing and 1 composite part: our fusion protein. All BioBricks are submitted with elaborate characterisation so the user may choose the best protein for their project accordingly. Our favourite basic part is BBa_K2643001 and our favourite composite part is BBa_K2643000.
| Fave Part | Name | Type | Description | Designer |
|---|---|---|---|---|
| ♥ | BBa_K2643000 | Composite | dxCas9-linker-Tn5 fusion protein (HIS Tag) | TU Delft 2018 |
| ♥ | BBa_K2643001 | Basic | dxCas9 (HIS Tag) | TU Delft 2018 |
| ♥ | BBa_K2643002 | Basic | Tn5 Transposase (HIS Tagged) | TU Delft 2018 |
| BBa_K2643003 | Basic | Glycine Helical peptide Linker (GHL | TU Delft 2018 | |
| ♥ | BBa_K2643004 | Basic | Coding region (CDS) of human erythropoietin (EPO) hormone | TU Delft 2018 |
| BBa_K2643005 | Basic | Human EPO gene with artificial intron (615 bp) | TU Delft 2018 | |
| BBa_K2643006 | Basic | Human EPO gene with artificial intron (135 bp) | TU Delft 2018 | |
| BBa_K2643007 | Basic | Human EPO gene with 2 artificial introns (615 + 135 bp) | TU Delft 2018 | |
| BBa_K2643008 | Basic | Human EPO gene with mutation 1 (5 bp) | TU Delft 2018 | |
| BBa_K2643009 | Basic | Human EPO gene with mutation 2 (12 bp) | TU Delft 2018 | |
| BBa_K2643010 | Basic | gRNA expression cassette for CRISPR-targeting lacZ | TU Delft 2018 | |
| BBa_K2643011 | Basic | Mosaic Ends-flanked Kanamycin cassette and RFP | TU Delft 2018 | |
| BBa_K2643012 | Basic | T7p expressing sgRNA targeting the EPO coding sequence | TU Delft 2018 |