Composite

Part:BBa_K1149020

Designed by: Margarita Kopniczky   Group: iGEM13_Imperial_College   (2013-09-08)
Revision as of 22:26, 30 September 2013 by Rkelwick (Talk | contribs)

Promoter J23104 - RBS B0034 - amilCP

The chromoprotein from the coral Acropora millepora, amilCP, naturally exhibits strong color when expressed. The protein has an absorbance maximum at 588 nm giving it a blue/purple color visible to the naked eye, thereby requiring no instruments to observe. This is a composite part from BBa_K608002 and BBa_K592009. It is functionally similar to BBa_K592031 which has a J23101 promoter instead of the J23104 but contains the same RBS and amilCP.

LigationamilCPICL.JPG

Figure: amilCP ligation cloned into E. coli strain NEB 10. The blue coloration made it very easy to select the ligations with the insert from the transformation plate:

We were inspired by the beautiful blue coral colour and taught the bacteria how to write our name with it.

PlasticityBluePlate.JPG

Figure: Made from streaking overnight cultures of amilCP transformed MG1655


PlasticityBluePlate.JPG Made from streaking overnight cultures of amilCP transformed MG1655

In addition to this, we have shown that bacterial paintings are achievable with this amazing biobrick (blue): https://parts.igem.org/File:Originals_combined_small.jpg Blue colouration relates to amilCP transformed MG1655

We have observed change in colour of an Overnight culture with the construct. However, it was not as blue as the overnight cultures obtained by Uppsala 2011. Although it is clear from our characterisation data that amilCP does express at a high level.

AmilCP_cultureICL.jpg

We have verified the sequence and detected 3 mutations compared to the expected results. However, these are located after the prefix and at the scar site and do not affect the coding region or the function of the construct.

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
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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