Reporter

Part:BBa_K4365003

Designed by: Giorgio Gilioli   Group: iGEM22_TU_Dresden   (2022-09-30)


turboRFP Selection Device with constitutive promoter

This device originates from the modificaiton of the BBa_J04450 contained in the pSB1C00 level 0 plasmid the RFC1000 assembly standard.

The original BBa_J04455 encoded for an RFP device, a transcriptional unit for the expression of the mRFP1 reporter gene, used for red-white colony screening after RFC1000 assembly of biological parts. The expression of mRFP1 was under the control of the Lac operon promoter and can be activate using IPTG. This promoter is leaky and we have replaced it with the J23100 consitutive promoter and we have replaced the mRFP1 with the superbright and fast maturing turboRFP fluorescent protein.

We have measure the activity of this device by comparing it to the original BBa_J04455 in induced and uninduced state and to a modified version of the BBa_J04455 containing the J23100 and expressing mRFP1.

This device can be used to perform white-orange colony screening after integration of a part.



Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 683


Usage and Biology

Continuous measurement of bacterial cultures with various promoters (lac promoter J04450 with and without IPTG, constitutive J23100) and downstream reporter genes mRFP1 and turboRFP.



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