Part:BBa_K3122001

6xHis Tag (Adapted to AEGIS' lamB display system)

This part is derived from BBa_K1223006. It contains 6 histidine residues, and it has been adapted to AEGIS' lamB display system in order to be used as the heterologous peptide tag expressed.

To see the design of this part go to the Design tab.

Characterization

Characterization of this part has been conducted by building a transcriptional unit BBa_K3122004 and performing assays with nickel beads in order to check the interaction between them and its force. This transcriptional unit was assembled in pARK1 alpha plasmid including the following parts:

• BBa_K3122003: Promoter J23107 adapted to type IIS assembly
• BBa_K2656009: RBS  B0030 adapted to type IIS assembly
• BBa_K3122002: coding sequence. This composite part is built with BBa_K3122000 (LamB protein) and BBa_K3122001 (6xHis tag) creating our AEGIS' lamB display system exposing 6xHis Tag.
• BBa_K2656026: Terminator B0015 adapted to type IIS assembly

How to use it

AEGIS' lamB display system that we created can be used by future iGEM teams on their MoClo reactions with the desired tag to create level 1 constructs. The only requirement is to adapt the tag sequence to the new scars designed, as this part has been adapted.

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]

References

J. Kim, C. Valencia, R. Liu and W. Lin, "Highly-Efficient Purification of Native Polyhistidine-Tagged Proteins by Multivalent NTA-Modified Magnetic Nanoparticles", Bioconjugate Chemistry, vol. 18, no. 2, pp. 333-341, 2007. Available:10.1021/bc060195l.