BioScaffol

BBa_K259002 Version 1

Component

Source:
http://parts.igem.org/Part:BBa_K259002
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
igem#experience
None
 
igem#sampleStatus
It's complicated
igem#status
Planning
 
synbiohub#ownedBy
user/james
 
synbiohub#ownedBy
user/myers
 
synbiohub#topLevel
BBa_K259002/1