Composite

Part:BBa_K1139020

Designed by: Naoki Watarai   Group: iGEM13_Tokyo_Tech   (2013-09-01)

PlacIq-M13-Plac-GFP on pSB3

We confirmed that M13 genome with two modifications related to our design kept plaque forming activity. One is replacement of the promoter for G2p (gen 2 protein) with a constitutive promoter, PlacIq (BBa_I14032). The other is accommodation of pSB3K3 backbone. Even though the plasmid has two different types of replication origins, M13 origin and pSB3 origin, this plasmid (BBa_K1139020) formed plaque(Fig. 1). In contrast, construction intermediates without a promoter upstream of g2p coding sequence (Promoterless-M13 + Plac, Promoterless-M13 + Plac-GFP BBa_K1139022) could not form plaque.

Fig. 1. PlacIQ-M13-Plac-GFP on pSB3 (BBa_K1139020)
Fig. 2. Promoterless-M13-Plac-GFP on pSB3 (BBa_K1139022)


















For more information, see [http://2013.igem.org/Team:Tokyo_Tech/Experiment/pSB-M13_Plasmid_Assay our work in Tokyo_Tech 2013 wiki].



Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 145
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 6071
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 7335


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Categories
Parameters
n/aPlacIq-M13-Plac-GFP on pSB3