Part:BBa_K1141002

Plac-RBS-KillerRed (IPTG-inducible)

KillerRed is a red fluorescent protein which produces ROS under green illumination (roughly 500-600 nm, absorption peak at 535 nm). This biobrick allows production of KillerRed under the control of the PLAC promoter (In this construction, actually a T5 phage promoter preceded by two lac operators for strong repression in presence of LacI).With this biobrick the production of KillerRed is IPTG-inducible. We used this part to trigger cell death in response to light illumination for controlling cell density in the Talk'E.coli project of Grenoble-EMSE-LSU 2013. KillerRed is a red fluorescent protein with Absorption in the green portion and emission in the red portion of the visible spectrum (figure 1):

Figure 1:The KillerRed protein absorption (left peak) and emission (right peak) spectra
Source: Detailed KillerRed description from Evrogen

Note that the information characterising this part was made with BBa_K1141001, which has the same sequence except for an additional Eco RI site between the Promoter and RBS. The vector for BBa_K1141001 is a high copy plasmid like pSB1C3 with ac col1 as duplication origin. KillerRed made by this biobrick dimerizes and isn't suitable for protein fusions.

Below is a SDS-PAGE gel showing purification of the protein, visible near the 31 kDa mark on the molecular weight ladder (figure 2):

Figure 2:KillerRed as can be obtained on an SDS-PAGE gel after purification. The protein stain can be clearly seen at the 31 kDa mark.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 133
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 289
  Illegal BsaI.rc site found at 580