Overview
Our best basic part is the coding sequence of the Cas3 Nuclease (BBa_K3096001), which is part of the Type 1 CRISPR/Cas adaptive system in E. coli. Unlike Cas9-based CRISPR systems, our system does not only cut, but also degrade the DNA targeted by the CRISPR arrays. We have amplified this part via PCR from genomic DNA of E. coli MG1655 and confirmed it via gel electrophoresis. With this part, we newly introduce the Cas3 nuclease to the iGEM registry. It is a nuclease bearing a lot of potential regarding biosafety and the knock-out of genes. For further information, please visit our Kill Switch page (Project/Kill Switch) and the iGEM registry of BBa_K3096001.
Basic Parts
| Parts/Name | Type | Collection | Length [bp] |
|---|---|---|---|
| BBa_K3096001 The Cas3 Nuclease is part of the Type I CRISPR/Cas system in E. coli |
Coding | Kill Switch | 2667 |
| BBa_K3096000 RBS |
RBS | 15 | |
| BBa_K3096006 NagC regulated promoter |
Regulatory | Kill Switch | 200 |
| BBa_K3096011 CAP promoter without LacI binding site |
Regulatory | Exendin-4 expression system | 183 |
| BBa_K3096050 Mnt |
Regulatory | Kill Switch | 90 |
| BBa_K3096014 CRISPR Array targeting Genomic E. coli DNA |
RNA | Kill Switch | 206 |
| BBa_K3096052 CRISPR Array targetting the Plasmid DNA of the glucose dependent Exendin-4 expression system |
RNA | Kill Switch | 147 |
| BBa_K3096019 6x His Tag |
Tag | CPP | 18 |
| BBa_K3096007 Type I CRISPR/Cas system surveillance complex Cascade ABCDE |
Coding | Kill Switch | 4351 |
| BBa_K3096013 Penetratin - a cell penetrating peptide |
Coding | CPP | 48 |
| BBa_K3096010 YTA2 - cell penetrating peptide |
Coding | CPP | 48 |
| BBa_K3096015 pVEC - a cell penetrating peptide |
Coding | CPP | 54 |
| BBa_K3096016 Tat-LK15 - improved Cell penetrating peptide |
Coding | CPP | 78 |
| BBa_K3096017 SynB1 - cell penetrating peptide |
Coding | CPP | 54 |
| BBa_K3096018 NagC repressor |
Coding | Kill Switch | 1373 |
| BBa_K3096023 GFP with linkers for fusion with His Tag and CPPs |
Coding | CPP | 726 |
| BBa_K3096030 Exendin-4 (GLP-1 analogue) with secretion signal |
Coding | Exendin-4 expression system | 126 |
