Team:Groningen/Parts
Parts
Our months of lab work contributed to the construction of several new parts. Please consult the registry for a more detailed explanation of our parts and the validation work that was performed for it.
Best Basic Part: dCas9 VRER
Our favorite part and the Groningen contender for Best New Basic part is dCas9 VRER. This variation on our improved spdCas9s for bacterial use, is adapted in such a way that it recognizes different PAM sites. These different sites ensure that the two Cas-complexes of our system do not compete for the binding sites of their counterpart. The difference in PAM sites ensures that the dCas9 VRER protein only binds to the sites which are specifically located on our pre-programmed reporter plasmid. It also prevent it from binding on the injected phage DNA. This also works the other way around as hCas9 only binds on the infecting phage DNA and not on the reporter plasmid, as it would cleave the plasmid upon binding to one of its sites.
Parts list
| .gb file | Part Number | Type | Part Name | Author | bp | ||
|---|---|---|---|---|---|---|---|
 |  | BBa_K2361001 | Coding | dCas9 VRER | Mart Bartelds | 4108 | |
| BBa_K2361000 | Coding | spdCas9 | Mart Bartelds | 4120 | ||
| BBa_K2361003 | RNA | pUSP45 tracrRNA | Thijs Ettema | 474 | ||
| BBa_K2361004 | RNA | CRISPR array | Sebald Verkuijl | 639 | ||
| BBa_K2361005 | RNA | CRISPR array containing a 20 spacer | Sebald Verkuijl | 691 | ||
| BBa_K2361006 | RNA | CRISPR array containing a 21 spacer | Sebald Verkuijl | 691 | ||
| BBa_K2361007 | RNA | CRISPR array containing empty spacers | Sebald Verkuijl | 625 | ||
| BBa_K2361009 | Regulatory | Nisin inducable lactococcal promoter | Thijs Ettema | 198 | ||
 | | BBa_K2361002 | Coding | hCas9 operon | Thijs Ettema | 6060 | |
| | BBa_K2361008 | Regulatory | p32 lactococcal promoter | Thijs Ettema | 196 | |
| | BBa_K2361010 | Regulatory | pUSP45 lactococcal promoter | Thijs Ettema | 606 |
