BioBrick type IIS subcloning
Upload your BioBrick sequence and subclone it into a type IIS-compatible vector
Leverage computer-aided design to choose the right fusion sites for your BioBrick
Yes, our computer-aided DNA designer is included in your free account
You can use one of ours lab-validated type IIS-compatible vectors
Our type IIS-compatible vectors come with fusion sites you can choose from
When done with the design you can directly ask for a free quotation for the synthesis
How does it work?
Since 2019, iGEM allows teams to subclone and submit their BioBricks using type IIS restriction enzymes. Thus, assembly methods such as GoldenGate, MoClo and LOOP are not accepted assembly standards for iGEM along with BioBrick RFC Standard and 3A assembly.
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.
A Type IIS assembly method uses Type IIS restriction enzymes, which are offsite cutters; they cut DNA at a specific distance from their recognition site. This allows for specified overhangs left by the Type IIS enzymes, which when intentionally designed can allow for multiple DNA fragments to be assembled together in the correct order and orientation.
iGEM currently accepts BsaI and SapI type IIS restriction sites coupled to 5 fusion sites specific for well-defined biological functions.
|5' BsaI site||5' Fusion site||BioBrick Type||3' Fusion site||3' BsaI site|
More info on the dedicated iGEM's webpage here.
A Modular Cloning System for Standardized Assembly of Multigene Constructs. Weber E, Engler C, Gruetzner R, Werner S, Marillonnet S. PLoS One. 2011. Feb 18;6(2):e16765. here.
A Golden Gate Modular Cloning Toolbox for Plants. Engler C, Youles M, Grützner R, Ehnert T-M, Werner S, Jones JDG, Patron NJ, Marillonnet S. ACS Synth. Biol. 2014. here.
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