Part:BBa_K2922031

T7 promoter-RBS(B0034)_kil(Ekil)

Summary

The usage of T7 promoter and RBS is to highly express the Lysis protein for Colicin-E1.

Identification

When we are building this circuit, we are doing the nucleic acid gel electrophoresis experiment to verify. After the circuit is built, we send the plasmid to sequence, and get the correct sequencing. After new molecular cloning experiments, we do Enzyme-Cut identification to certify the plasmid is correct. We use the EcoRI and PstI to cut the plasmid, then we get the target separate fragment-284bp. (Fig.1)

Fig.1 The result of this plasmid cut with enzyme EcoRI and PstI.

The plasmids are transformed into BL21(DE3).After growing the single colonies,SDS-PAGE protein gel electrophoresis is used to certify the expression of Ekil,then we get the target molecular weight-4.8kDa.(Fig.2)

Fig.1 The result of the protein molecular weight.

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
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 176
  
• 1000
  COMPATIBLE WITH RFC[1000]