Part:BBa_K5403002

Flexible GSGSGSGSAS linker with NheI restriction site

The GSGSGSGSAS linker can be used to fuse two proteins together by recombinant expression. It provides flexibility to prevent that the fusion of the two distinct proteins would cause misfolding. Additionally, it provides an NheI restriction site that can be used to exchange the gene for the C-terminal protein with a different one for easy cloning. The part was used in combination with other genes within the plasmids pET28a (expression in E. coli) and pCHERRY3 (expression in m. smegmatis). In these plasmids, the NheI restriction site is unique.

Part BBa_K5403002 was used to create the fusion proteins encoded by BBa_K54030012 - BBa_K54030020. Protein expression results are available for parts BBa_K54030012 and BBa_K54030013. The part was used in combination with other genes within the plasmids pET28a (expression in E. coli) in parts BBa_K5403012 - BBa_K5403013 and pCHERRY3 (expression in m. smegmatis) in parts BBa_K5403014 - BBa_K5403020. In these plasmids, the NheI restriction site is unique. pCHERRY3 was a gift from Tanya Parish (Addgene plasmid # 24659 ; http://n2t.net/addgene:24659 ; RRID:Addgene_24659).

pCHERRY3 was a gift from Tanya Parish (Addgene plasmid # 24659 ; http://n2t.net/addgene:24659 ; RRID:Addgene_24659).

References

Addgene plasmid # 24659. Sensitive detection of gene expression in mycobacteria under replicating and non-replicating conditions using optimized far-red reporters. Carroll P, Schreuder LJ, Muwanguzi-Karugaba J, Wiles S, Robertson BD, Ripoll J, Ward TH, Bancroft GJ, Schaible UE, Parish T. PLoS One. 2010 . 5(3):e9823. 10.1371/journal.pone.0009823 PubMed 20352111

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]