Project
Binding

Part:BBa_K1378009

Designed by: Zhang Zijian & Meng Liuyi   Group: iGEM14_Peking   (2014-10-06)

INPNC-Linker (rTEV)-MVN & GFP

Introduction

This plasmid is an advanced testing plasmid for our E. coli - M. aeruginosa binding assay. It differs with the part BBa_K1378007 in that we added a special linker between INPNC and MVN. We inserted two transcription units into one plasmid and they are expressed under the promoters J23105 and J23117 respectively. After this plasmid is transformed into E. coli, the mannan binding protein MVN (microvirin) will be displayed on the cell surface by INPNC (Ice Nucleation Protein N and C terminal) and the E. coli cell will express GFP as well. There is a linker between INPNC and MVN that can be specifically digested by rTEV protease so we can add rTEV protease to control the binding activity of our E. coli cells. The cell culture with rTEV protease added can serve as the control group for the binding assay.


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
    Illegal NheI site found at 1528
    Illegal NheI site found at 1551
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 466
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2229
    Illegal SapI site found at 1071

Characterization

We did not carry out the binding assay for this part. Detailed characterization of INPNC-MVN can be found in the part BBa_K1378007.


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