Difference between revisions of "Part:BBa K259002"

Revision as of 18:01, 1 August 2009

BioScaffold Linker - Removes Stop Codons & scars & replaces with a Gly-Ser linker

This part is part of a family of Bioscaffold - Linkers.

This part will help users interested in protein-fusion. The usage of the part will allow in frame assembly of 2(+) proteins by:

• Removing the stop codons from the upstream protein.
• Removing the scar created by the Assembly Standard 10 (RFC10) construction of 2(+) protein coding sequences.
• Maintaining the start codon of the downstream protein.
• Adding a flexible (Gly-Ser-Gly) linker between the two protein coding sequences.
• Acting as a translational buffer if ligation is incorrect.

Assembly Instructions

• Create an Assembly Standard 10 fusion as below ;

 **Upstream Protein Part - Bioscaffold Linker - Downstream Protein Part**

• Use the BioScaffold Specific Enzymes

(i)Restrict with BpuEI to remove upstream scar (created between Upstream Protein - Bioscaffold) and stop codons from upstream protein part.
(ii)Ligate and you will get the intermediate product;

 **Upstream Protein (w/out Stop codons) - Tyr - BioScaffold Linker - Downstream Protein**

• Restrict with BseRI to remove downstream scar.
• Ligate to get the final product;

  **Upstream Protein - Tyr-Gly-Ser-Gly- Downstream Protein**

Sequence and Features