Team:TUDelft/Parts
When constructing our parts, we made use of the IDT synthesis offer. This way we were able to generate our required parts in a fast and efficient way. All of the parts we synthesised contain the coding sequence, RBS, promoter and terminators, if applicable. Therefore, these parts are composite. The different basic parts comprising the composite parts 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 and restriction ligation on the synthesised composite parts. We created a part collection comprised of 4 composite parts encoding for different Tardigrade Proteins (TDPs) and the 4 basic parts of these genes, all 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.
Improved BioBrick / Further Characterisation
In our project, we added to the characterisation of an existing Biobrick, the TolR, periplasmic domain design by Gavin Sutton from iGEM Australia 2016. Additionally, our TorA-GFP composite part was based on the BioBrick created by NTU Trontheim with some alterations, such as the removal of the RFP sequence. This enabled us to build on their knowledge and improve the function.
All Parts
| Fave Part | Name | Type | Description | Designer | Length |
|---|---|---|---|---|---|
| BBa_K2306000 | Coding | TorA-GFP fusion | Gabriella Tany | 846 | |
| BBa_K2306001 | Composite | TorA-GFP fusion (LacI promoter) | Gabriella Tany | 1212 | |
| BBa_K2306002 | Coding | Secretory-abundant heat soluble protein 68234 (SAHS 68234) | Guillermo Serena Ruiz | 525 | |
♥ |
BBa_K2306003 | Coding | Secretory-abundant heat soluble protein 33020 (SAHS 33020) | Guillermo Serena Ruiz | 509 |
| BBa_K2306005 | Coding | Cytosolic-abundant heat soluble protein 106094 (CAHS 106094) | Guillermo Serena Ruiz | 684 | |
| BBa_K2306006 | Coding | Cytosolic-abundant heat soluble protein 94205 (CAHS 94205) | Guillermo Serena Ruiz | 681 | |
| BBa_K2306007 | Composite | Secretory-abundant heat soluble protein 68234 (SAHS 68234) (T7 promoter, RBS and double terminator) | Guillermo Serena Ruiz | 768 | |
| ♥ | BBa_K2306008 | Composite | Secretory-abundant heat soluble protein 33020 (SAHS 33020) (T7 promoter, RBS and double terminator) | Guillermo Serena Ruiz | 752 |
| BBa_K2306010 | Composite | Cytosolic-abundant heat soluble protein 106094 (CAHS 106094) (T7 promoter, RBS and double terminator | Guillermo Serena Ruiz | 927 | |
| BBa_K2306011 | Composite | Cytosolic-abundant heat soluble protein 94205 (CAHS 94205) (T7 promoter, RBS and double terminator) | Guillermo Serena Ruiz | 924 | |
| BBa_K2306012 | Coding | Cas13a | Jeroen Jacques | 3920 | |
| BBa_K2306013 | Composite | Cas13a spacer with flanking double repeats, constitutive promoter | Jeroen Jacques and Jasper Veerman | 134 | |
| BBa_K2306014 | RBS | RBS from bacteriophage T7 gene 10 and Shine Dalgarno | Guillermo Serena Ruiz | 30 | |
| BBa_K2306015 | Other | Cas13a spacer with flanking double repeats | Jeroen Jacques and Jasper Veerman | 99 |
Basic Parts
| Fave Part | Name | Type | Description | Designer | Size (bp) |
|---|---|---|---|---|---|
| BBa_K2306002 | Basic | Secretory-abundant heat soluble protein (SAHS) 68234 | Guillermo Serena Ruiz | 525 | |
| ♥ | BBa_K2306003 | Basic | Secretory-abundant heat soluble protein (SAHS) 33020 | Guillermo Serena Ruiz | 509 |
| BBa_K2306005 | Basic | Cytosolic-abundant heat soluble protein (CAHS) 106094 | Guillermo Serena Ruiz | 684 | |
| BBa_K2306006 | Basic | Cytosolic-abundant heat soluble protein (CAHS) 94205 | Guillermo Serena Ruiz | 681 | |
| BBa_K23006000 | Basic | TorA GFP | Gabriella Tany | 681 | |
| BBa_K2306012 | Basic | Cas13a | Jeroen Jacques | 3456 | |
| BBa_K2306014 | RBS | RBS from bacteriophage T7 gene 10 and Shine Dalgarno | Guillermo Serena Ruiz | 30 | |
| BBa_K2306015 | Basic | Spacer with flanking double repeats for Cas13a | Jeroen Jacques, Jasper Veerman | 99 |
Composite Parts
| Fave Part | Name | Type | Description | Designer | Size (bp) |
|---|---|---|---|---|---|
| BBa_K2306007 | Composite | Secretory-abundant heat soluble protein (SAHS) 68234 with inducible T7 promoter, RBS and double terminator | Guillermo Serena Ruiz | 833 | |
| ♥ | BBa_K2306008 | Composite | Secretory-abundant heat soluble protein (SAHS) 33020 with inducible T7 promoter, RBS and double terminator | Guillermo Serena Ruiz | 817 |
| BBa_K2306010 | Composite | Cytosolic-abundant heat soluble protein (CAHS) 106094 with inducible T7 promoter, RBS and double terminator | Guillermo Serena Ruiz | 992 | |
| BBa_K2306011 | Composite | Cytosolic-abundant heat soluble protein (CAHS) 94205 with inducible T7 promoter, RBS and double terminator | Guillermo Serena Ruiz | 989 | |
| BBa_K2306001 | Composite | TorA GFP fusion with inducible LacI promoter, RBS and double terminator | Gabriella Tany | 681 | |
| BBa_K2306015 | Composite | Cas13a spacer with flanking double repeats, constitutive promoter | Jeroen Jacques, Jasper Veerman | 134 |
Part Collection
| Fave Part | Name | Type | Description | Designer | Size (bp) |
|---|---|---|---|---|---|
| BBa_K2306007 | Composite | Secretory-abundant heat soluble protein (SAHS) 68234 with inducible T7 promoter, RBS and double terminator | Guillermo Serena Ruiz | 833 | |
| ♥ | BBa_K2306008 | Composite | Secretory-abundant heat soluble protein (SAHS) 33020 with inducible T7 promoter, RBS and double terminator | Guillermo Serena Ruiz | 817 |
| BBa_K2306010 | Composite | Cytosolic-abundant heat soluble protein (CAHS) 106094 with inducible T7 promoter, RBS and double terminator | Guillermo Serena Ruiz | 992 | |
| BBa_K2306011 | Composite | Cytosolic-abundant heat soluble protein (CAHS) 94205 with inducible T7 promoter, RBS and double terminator | Guillermo Serena Ruiz | 989 | |
| BBa_K2306002 | Basic | Secretory-abundant heat soluble protein (SAHS) 68234 | Guillermo Serena Ruiz | 525 | |
| ♥ | BBa_K2306003 | Basic | Secretory-abundant heat soluble protein (SAHS) 33020 | Guillermo Serena Ruiz | 509 |
| BBa_K2306005 | Basic | Cytosolic-abundant heat soluble protein (CAHS) 106094 | Guillermo Serena Ruiz | 684 | |
| BBa_K2306006 | Basic | Cytosolic-abundant heat soluble protein (CAHS) 94205 | Guillermo Serena Ruiz | 681 |
