Part:BBa_K936021

Cutinase and polyhistidine tag

LC Cutinase coding sequence with polyhistidine tag attached. The polyhistidine tag allows our research team to perform assays to discover where our enzyme is being released, both in the media and in the cell.

Through p-nitrophenyl butyrate (pNPB) assays, the UC Davis team gathered enough data to determine that the LC-Cutinase part (BBa_K936000) incorporated in this part most likely exhibits its intended function as an esterase. A detailed description of these assays can be found on the Module Engineering Project page: http://2012.igem.org/Team:UC_Davis/Project/Catalyst.
Additionally, the following data is further described on the UC Davis Cutinase Activity data page: http://2012.igem.org/Team:UC_Davis/Data/Cutinase_Activity

(Above Graph) Results of pNPB assay detailing the constitutive construct as having the highest esterase activity.
(Above Graphs) The graphs above detail higher activity among the constitutive construct and some mutant variants in esterase activity compared to the control and wild type per unit cell.

(Above graph) This graph represents a possible production of cutinase by the cells upon entering stationary phase.

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 600
  
• 1000
  COMPATIBLE WITH RFC[1000]