Translational_Unit

Part:BBa_K2387054

Designed by: José Manuel Salvador López   Group: iGEM17_Wageningen_UR   (2017-10-18)
Revision as of 11:19, 31 October 2017 by Josemasl (Talk | contribs)


mRFP controlled by inducible araC/pBAD promoter

mRFP is a red monomeric fluorescent protein. In this composite, its expression is regulated by the araC/pBad promoter and by a strong RBS. mRFP presents a maximum pick of excitation at 584 nm and a maximum pick of emission at 607 nm. mRFP was engineered from DsRed to grant it a monomeric conformation.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1205
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1144
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 979
    Illegal AgeI site found at 1793
    Illegal AgeI site found at 1905
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 961


Functional Parameters

In order to test the functionality of this protein under the araC/pBad promoter, the absorbance and the fluorescence spectra were measured after induction with 0.02% arabinose (Figure 1).

Figure 1: Absorbance and fluorescence spectra of mRFP, measured after induction with 0.02% arabinose.

This protein is part of a collection of split proteins developed for BiFC analysis and tested with the CpxR system (BBa_K2387032). These proteins were characterized through multiple experiments using as interacting proteins antiparallel synthetic leucine zippers. The proteins of this collection are the ones found in Table 1.

Table 1: Split Proteins.
Protein Part Number (Full length) Part Number (Split Protein)
mRFP BBa_K2387054 BBa_K2387055
eYFP BBa_K2387003 BBa_K2387065
mVenus BBa_K2387045 BBa_K2387046
sfGFP BBa_K2387047 BBa_K2387048
mCerulean BBa_K2387052 BBa_K2387053
[edit]
Categories
//cds/reporter
//cds/reporter/rfp
//chassis/prokaryote
//function/reporter/fluorescence
Parameters
absMax. 515 nm
chassisE. coli
colorRed Fluorescence, Bright Pink cells
emissionMax. 607 nm