Part:BBa_K1139022

Promoterless-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 (gene 2 protein) with a constitutive promoter, PlacI^q (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. 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 100
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 6026
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 7287