Help:Standards/Assembly/Type IIS
Motivation & Discussion
With a Type IIS assembly method, multiple parts can be assembled easily and reliably in a single reaction, and this can be scaled further to create more complex genetic circuits quickly, efficiently, and affordably.
Currently, there are several Type IIS assembly standards in use, many of these were designed for specific purposes. iGEM believes strongly in standards, and as we approach Type IIS assembly methods we want to support one that we believe will be useful, reliable, long-lasting and widespread for the synthetic biology community.
We want to ensure that the assembly standard and method we support is widely used, tested, and has flexibility for different applications and groups.
Technical Specifications
Compatibility/Illegal Sites
iGEM Type IIS compatible parts must not contain the Bsa1 and Sap1 Type IIS restriction enzyme sites, as these are required for assembly. Parts that have these “illegal” restriction sites are not compatible with the standard.
| Enzyme | Type | Sequence |
|---|---|---|
| BsaI | Illegal | GGTCTC |
| SapI | Illegal | GCTCTTC |
Fusion Sites
| Fusion Site 5' | Part | Fusion Site 3' |
|---|---|---|
| GGAG | Promoter | TACT |
| TACT | 5'UTR | AATG |
| AATG | CDS | GCTT |
| GCTT | CDS | CGCT |
Accepted Standards: BioBrick RFC[10] | iGEM Type IIS RFC[1000]