Part:BBa_K2448004

mCherry

This codon optimized mCherry for Escherichia coli K12 is derived from (BBa_J06504). The optimization has been performed with help of [http://www.jcat.de/ JCAT].

Before optimization, the CAI-Value of the original sequence was 0.387 with a 62% GC-Content. The optimization allows a CAI-Value for the improved sequence of 1.0 with a 47% GC-Content. NcoI and BbsI restrictions sites have also been removed to allow potential Golden Gate Assembly use and complementarity with the Universal Biosensing Chassis (BBa_K2448023 and BBa_K2448024)

Usage and Biology

mCherry is a fluorescent monomeric protein widely used in biotechnology. Like the majority of red fluorescent proteins, it is derived from the Red Fluorescent protein from Discosoma sp. mCherry is sometimes preferred over other fluorophores due to its color, its rapid maturation time (t0.5=15 min), its lower brightness (around 50% of eGFP) as well as its improved photostability and resistance to bleaching. Figure 1 illustrates the intense colour of mCherry expressing E. coli cells.

Applications are various: Fusions, promoter & secretion studies, FRET and other quantitative imaging techniques.

Our team used this codon optimized mCherry to build the Universal Biosensing Chassis (BBa_K2448023 and BBa_K2448024) and test our Psicose Biosensors (BBa_K2448025, BBa_K2448026, BBa_K2448027, BBa_K2448028, BBa_K2448029, BBa_K2448030 and BBa_K2448031) as well as our Fructose Biosensor (BBa_K2448032).

Figure 1. mCherry expression in E. coli cells harbouring BBa_K2448032 in the presence of Fructose.

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 570
  Illegal AgeI site found at 682
  
• 1000
  COMPATIBLE WITH RFC[1000]