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

BioBrick cloning with type IIS


Key benefits

Free

Yes, our computer-aided DNA designer is included in your free account

Lab Validated

You can use one of ours lab-validated type IIS-compatible vectors

iGEM-ready

Our type IIS-compatible vectors come with fusion sites you can choose from

Synthesis-ready

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[10] 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
GGTCTC GGAG
(Type A)		 Promoter TACT
(Type B)		 GAGACC
GGTCTC TACT
(Type B)		 5UTR AATG
(Type C)		 GAGACC
GGTCTC AATG
(Type C)		 CDS GCTT
(Type E)		 GAGACC
GGTCTC GCTT
(Type E)		 Terminator CGCT
(Type F)		 GAGACC

Reference:
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.

Disclaimer:
BioBrick® is a registered trademark of BioBricks Foundation

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BioBrick cloning with type IIS-logo
Accepted inputs: A, C, G, T, W (weak: A, T), S (strong: C, G ), M (amino: A, C), K (keto: G, T), R (purine: A, G ), Y (pyrimidine: C, T), B (not A), D (not C), H (not G), V (not T), N (any basebase)
Blast BP: 0
Sequence and name fields will be overridden
Biosafety level*
By selecting the biosafety level corresponding to the project, customer confirms that (i) such classification is correct and accurate and (ii) that the customer has read, understood and accepted the biosafety confirmation.

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