Difference between revisions of "Part:BBa K1139020"
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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.
  
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[[Image:Titech2013_parts_K1139020_Fig1.jpg|thumb|left|500px|'''Fig. 1.''' PlacIQ-M13-Plac-GFP on pSB3 (BBa_K1139020)]]
+
[[Image:Titech2013_parts_K1139020_Fig1.jpg|thumb|left|1000px|'''Fig. 1.''' PlacIQ-M13-Plac-GFP on pSB3 (BBa_K1139020)]]
  
−
[[Image:Titech2013_parts_K1139020_Fig2.jpg|thumb|left|500px|'''Fig. 2.''' Promoterless-M13-Plac-GFP on pSB3 ([https://parts.igem.org/Part:BBa_K1139022 BBa_K1139022])]]
+
[[Image:Titech2013_parts_K1139020_Fig2.jpg|thumb|left|1000px|'''Fig. 2.''' Promoterless-M13-Plac-GFP on pSB3 ([https://parts.igem.org/Part:BBa_K1139022 BBa_K1139022])]]
  
 
<i>GFP</i> expression is regulated by LacI repressor.
 
<i>GFP</i> expression is regulated by LacI repressor.

Revision as of 16:29, 25 September 2013

PlacIq-M13-Plac-GFP on pSB3

PlacIQ is a constitutive promoter.

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)

GFP expression is regulated by LacI repressor.

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