Part:BBa_K1493803
promoter with CIλ and LacI operator sites
Promoter designed to have operators sites for the Lac repressor and the cI lambda repressor, based on sequences in Elowitz et. al. 2007
This promoter contains two LacI operator sites at the core and proximal position (middle and right part of the promoter) and two cI lambda operator sites at the distal site (left part of the promoter).
Usage and Biology
The promoter was combined with GFP to check for its functionality.
Figure 1: plates with the six diferent promters, that where created for BananaGuard, upstream of GFP. P1 is BBa_K1493801, P2 is BBa_K1493802, P3 is BBa_K1493803, P4 is BBa_K1493804, P5 is BBa_K1493805, P6 is BBa_K1493806.
Figure 1 shows the P3 (promoter with CIλ and LacI operator sites), it is not expressing GFP suggesting that it is either non functional or a very weak promoter.
This Promoter is part of a promoter set used in [http://2014.igem.org/Team:Wageningen_UR BananaGuard] listed here:
Contribution
Group: Team Jiangnan 2018.
Author: Yini Luo.
Summary: We combined rfp to promoter BBa_K1493803 to measure its strength and compared it with another 5 promoters from iGEM14_Wageningen_UR. This part is used for the characterization of the promoter BBa_K1493803.
BBa_K1493803 contains two LacI operator sites at the core and proximal position (middle and right part of the promoter) and two cI lambda operator sites at the distal site (left part of the promoter).We transduced the recombinant plasmid containing the target promoter and an rfp reporter gene into E.coli. In this experiment, we chose BioTek Synergy multifunctional enzyme mark to measure the fluorescence intensity and OD450, PBS was used as the blank control. According to the optimum detection wavelength of RFP, the parameters for detecting fluorescence intensity were: excitation wavelength of 584nm and emission wavelength of 607nm. The fluorescence / OD450 was used to measure the RFP expression level of bacteria. The results are provided in Figure 1.For details, please click BBa_K2568012.
Figure 1.Comparisions of the expression of 6 promoters. Expression of the promoters are ordered from high to low. Expression of the blank control is considered 0. The promoter 3 is considered unexpressed as no fluorescence expression was detected.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
| None |