Composite

Part:BBa_K1682017

Designed by: iGEM15_HKUST-Rice   Group: iGEM15_HKUST-Rice   (2015-09-15)

ParaBAD-GFP-Plac-RFP

This composite part is consists of a L-arabinose inducible ParaBAD with GFP reporter BBa_I13540, and an IPTG inducible Plac with RFP reporter Part:BBa_K1682016. The part was designed to investigate the possible effect of parallel sensing within a single plasmid.

Mechanism

Parallel expression BBa K1682017.jpg

Figure 1. Schematic diagram of the inducible ParaBAD-GFP-Plac-mrfp construct. The system can sense L-arabinose to give GFP output or sense IPTG to give RFP output.

Usage and Biology

In order to investigate the effect of parallel sensing, the dose response of ParaBAD-gfp in pSB3K3-BBa_K1682017 (double construct) was compared with that in pSB3K3-BBa_I13540 (single construct) in E. coli DH10B.

Parallel expression BBa K1682017 fig2.jpg

Figure 2. Response curve of ParaBAD-gfp in a single construct and double construct under varying L-arabinose concentration.The different blue curves indicate different induction level of the co-expressed Plac-mrfp construct. At varying the induction level of the co-expressed Plac-mrfp construct seems to have an effect on the expression level of ParaBAD-gfp in a double construct. At a higher IPTG concentration, the maximum expression of ParaBAD-gfp decreases.


Sequence and Features

Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1205
    Illegal NheI site found at 3185
    Illegal NheI site found at 3208
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 4386
    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 2882
    Illegal AgeI site found at 2994
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1880


[edit]
Categories
//chassis/prokaryote/ecoli
Parameters
None