Regulatory

Part:BBa_K1493801

Designed by: Max van der Ploeg   Group: iGEM14_Wageningen_UR   (2014-10-07)

Promoter with Tet operator site

Promoter designed to have operators sites for the tetracycline repressor, based on sequences in Elowitz et. al. 2007

This promoter contains two TetR operator sites at the core and proximal position (middle and right part of the promoter)

Usage and Biology

The promoter was combined with GFP to check for its functionality. Wageningen_UR_killswitch_Pic13.jpg 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 P1 (Promoter with Tet operator site) expressing GFP, suggesting that it is a functional promoter.


This Promoter is part of a promoter set used in [http://2014.igem.org/Team:Wageningen_UR BananaGuard] listed here:

BBa_K1493802

BBa_K1493803

BBa_K1493804

BBa_K1493805

BBa_K1493806


Contribution

Group: Team Jiangnan 2018.
Author: Yini Luo.
Summary: We combined rfp to promoter BBa_K1493801 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_K1493801.
BBa_K1493801 contains two TetR operator sites at the core and proximal positions (middle and right parts 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_K2568010.

T--Jiangnan--Promoter.png

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


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]


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Categories
Parameters
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