Part:BBa_K1119006

CMV promoter

CMV (Cytomegalovirus) promoter is a constitutive mammalian promoter.

The partsregistry contains CMV promoter (BBa_J52034) submitted by Slovenia 2006 team and characterized by DTU-Denamrk 2011 team. In addition, there are five twins of this CMV promoter, including BBa_I712004 was submitted by Ljubljana 2007 team and characterized by Heidelberg 2009 team using BBa_K203100 pSMB_MEASURE (Promoter measurement plasmid). However, according to the user reviews, team LMU-Munich 2010 stated that this part is not a CMV promoter but rather a long version of lacI gene for prokaryotes. Although there were many uses of this promoter, our team decided to build a reliable CMV promoter for our constitutive glyoxylate shunt construct.

Characterization

In our characterization, the coding sequence of CMV promoter was assembled with GFP reporter (BBa_K648013) and hGH polyA terminator (BBa_K404108) using Freiburg’s RFC25 format.

The pCMV-GFP was then transfected into HEK293FT cells and in vivo green fluorescence signal was observed under confocal microscope.

The positive control was pEGFP-N1 (Clontech) that contains CMV promoter and EGFP reporter. A negative control was made by GFP generator that does not contain the CMV promoter.

The [http://2013.igem.org/Team:Hong_Kong_HKUST/characterization detailed protocol] of our characterization can be found in HKUST iGEM 2013 Wiki.

Figure 1. CMV promoter drives expression of GFP. HEK cells transfected with pCMV-GFP expressed GFP. Signal. HEK cells transfected with the commercial pEGFP-N1 showed similar results, while the negative control GFP without any promoter was not expressed.

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]