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 | DNA sequence_feature |
| Sequences | BBa_K259002_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 giving ;
**Upstream Protein Part - Bioscaffold Linker - Downstream Protein Part
*Use the BioScaffold Specific Enzymes
**Restrict with BpuEI to remove upstream scar (created between Upstream Protein - Bioscaffold) and stop codons from upstream protein part.
**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
Notes
Part was designed taking into account RFC10 assembly standard and as an attempt to be upwards compatible with RFC10.Source
This part was designed de novo. The sequence does not occur naturally.| Sequence Annotation | Location | Component / Role(s) |
| Tyr Residue involved in Restriction/Ligation BpuEI Site BpuEI Site Gly-Ser-Gly Linker Met involved in Restriction/Ligation Translational Buffer BseRI Site BseRI Site | 41,43 3,8 21,26 44,52 53,55 56,62 63,68 81,86 | feature/misc sequence_feature feature/misc sequence_feature feature/misc sequence_feature sequence_feature feature/dna feature/misc sequence_feature stop_codon feature/stop sequence_feature feature/misc feature/misc sequence_feature |