Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Petros Mina
Date created: 2009-07-31 11:00:00
Date modified: 2015-05-08 01:11:42
BioScaffold Linker - Removes Stop Codons & scars & replaces with a Gly-Ser linker
| Types | DnaRegion |
| Roles | sequence_feature DNA |
| Sequences | BBa_K259004_sequence (Version 1) |
Description
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**
[[Image:BCCS_Assembly_of_Scaffold.jpg|thumb|500px|center|The Product of Assembly between 2 proteins and the Bioscaffold Linker. ]]
*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 CspCI to remove downstream scar.
*Ligate to get the final product;
**Upstream Protein - Tyr-Gly-Ser-Gly- Downstream Protein**
[[Image:BCCS_Bioscaffold_Reaction_Explanation_CspCI.jpg|thumb|500px|center|The Reaction and the product. ]]
Notes
This part was designed taking into consideration the requirements by Assembly Standard 10 (RFC10).Source
This part is completely designed de novo. This sequence does not occur naturally.| Sequence Annotation | Location | Component / Role(s) |
| BpuEI BpuEI Tyr Involved in Restriction/Ligation Gly-Ser-Gly Linker Met Involved in Restriction/Ligation Translational Buffer CspCI (5' Recognition Seq) Translational Buffer CspCI (3' Recognition Seq) | 3,8 21,26 41,43 44,52 53,55 56,65 66,68 70,72 74,77 | feature/misc sequence_feature feature/misc sequence_feature sequence_feature feature/misc sequence_feature feature/dna sequence_feature feature/misc feature/stop stop_codon sequence_feature feature/misc stop_codon feature/stop feature/misc sequence_feature |