Difference between revisions of "Part:BBa K1139020"
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PlacIQ is a constitutive promoter.
 
PlacIQ is a constitutive promoter.
−
<i>GFP</i> expression is regulated by LacI repressor.
+
<i>GFP</i> expression is regulated by LacI repressor.<br>
 
We confirmed that M13 genome with two modifications related to our design kept plaque forming activity.  One is replacement of the promoter for g2p protein with a constitutive promoter, PlacIq ([https://parts.igem.org/Part:BBa_I14032 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 ([https://parts.igem.org/Part:BBa_1139020 BBa_K1139020]) formed plaque.  In contrast, construction intermediates without a promoter for g2p coding sequence (Promoterless-M13 + Plac, Promoterless-M13 + PlacGFP [https://parts.igem.org/Part:BBa_1139022 BBa_K1139022]) could not form plaque.
 
We confirmed that M13 genome with two modifications related to our design kept plaque forming activity.  One is replacement of the promoter for g2p protein with a constitutive promoter, PlacIq ([https://parts.igem.org/Part:BBa_I14032 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 ([https://parts.igem.org/Part:BBa_1139020 BBa_K1139020]) formed plaque.  In contrast, construction intermediates without a promoter for g2p coding sequence (Promoterless-M13 + Plac, Promoterless-M13 + PlacGFP [https://parts.igem.org/Part:BBa_1139022 BBa_K1139022]) could not form plaque.
  

Revision as of 16:33, 25 September 2013

PlacIq-M13-Plac-GFP on pSB3

PlacIQ is a constitutive promoter. GFP expression is regulated by LacI repressor.
We confirmed that M13 genome with two modifications related to our design kept plaque forming activity. One is replacement of the promoter for g2p 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. In contrast, construction intermediates without a promoter for g2p coding sequence (Promoterless-M13 + Plac, Promoterless-M13 + PlacGFP 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)













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