Part:BBa_K1123017

DNA Coding Sequence for EGFP Protein

This part contains the DNA sequence for the EGFP protein. It can be placed behind any promoter of your choice and expressed with ease. Within the scope of our project this protein was expressed as a control. It is a highly fluorescent protein and could be easily analysed to reveal the workings of your promoter or other constructs.

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]

Characterization

For the iGEM 2013 Purdue team we characterized our parts according to their characterization datasheets. The data sheet for this particular biobrick can be found here.

Characterization by TU-Eindhoven 2013

EGFP is a very standart part to use in the lab. However it was used within the scope of our project as a control to ensure that our expression was being performed correctly. Of course a lot of cloning work was performed with this part but for the characterization only the protein expression is of real importance, especially within our project.

After protein expression had taken place the solution was spun down and bugbustered. A small portion of the resulting supernatant and pellet was then taken and loaded onto an SDS gel with the following result:

Here it is obvious that the expression was performed extremely well and that the protein could be found in both the supernatant as well as in the pellet itself, showing just how much had been created.

No further tests were performed with this particular part, although it was itself used in another composite part: BBa_K1123005.