Difference between revisions of "Part:BBa K259002"
Line 17: | Line 17: | ||
**Upstream Protein Part - Bioscaffold Linker - Downstream Protein Part** | **Upstream Protein Part - Bioscaffold Linker - Downstream Protein Part** | ||
+ | |||
+ | |||
+ | |||
+ | [[Image:BCCS_Assembly_of_Scaffold.jpg|thumb|500px|center|The Product of Assembly between 2 proteins and the Bioscaffold Linker. ]] | ||
+ | |||
Line 30: | Line 35: | ||
**Upstream Protein - Tyr-Gly-Ser-Gly- Downstream Protein** | **Upstream Protein - Tyr-Gly-Ser-Gly- Downstream Protein** | ||
+ | |||
+ | [[Image:BCCS_Bioscaffold_Reaction_Explanation.jpg|thumb|500px|center|The Reaction and the product. ]] | ||
Revision as of 18:46, 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
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]