Difference between revisions of "Part:BBa K1416000"
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This part was used by the UT Austin 2014 iGEM team in their [http://2014.igem.org/Team:Austin_Texas/kit#Results_and_Data ncAA kit project].  In this work they demonstrated that while ONBY itself is slightly toxic to the cells, the synthetase/tRNA pair acted with good fidelity at incorporating only ONBY.  Additionally, this part was also used in the UT Austin 2014 iGEM [http://2014.igem.org/Team:Austin_Texas/photocage Photocage project], where the part successfully incorporated ONBY within T7 RNA polymerase (RNAP), yielding a light-activated T7 RNAP.
 
This part was used by the UT Austin 2014 iGEM team in their [http://2014.igem.org/Team:Austin_Texas/kit#Results_and_Data ncAA kit project].  In this work they demonstrated that while ONBY itself is slightly toxic to the cells, the synthetase/tRNA pair acted with good fidelity at incorporating only ONBY.  Additionally, this part was also used in the UT Austin 2014 iGEM [http://2014.igem.org/Team:Austin_Texas/photocage Photocage project], where the part successfully incorporated ONBY within T7 RNA polymerase (RNAP), yielding a light-activated T7 RNAP.
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[[File:IodoY-UT Austin 2014.png|thumb]]
  
 
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Revision as of 01:00, 17 October 2014

The tRNA synthetase/tRNA needed for incorporating o-(2-nitrobenzyl)-L-tyrosine (ONBY) at a UAG codon

The part includes the gene coding for the Methanocaldococcus jannaschii synthetase mutated to charge the orthogonal tRNA (also in the part) with the non-canonical amino acid o-nitrobenzyl tyrosine (ONBY) along with the proper promoters and terminator. This part encodes for the incorporation of ONBY during translation at AUG amber stop codons in a cell with Release Factor 1 (RF1) knocked out such as "Amberless" E. coli or "RF0" E. coli.

This part was used by the UT Austin 2014 iGEM team in their [http://2014.igem.org/Team:Austin_Texas/kit#Results_and_Data ncAA kit project]. In this work they demonstrated that while ONBY itself is slightly toxic to the cells, the synthetase/tRNA pair acted with good fidelity at incorporating only ONBY. Additionally, this part was also used in the UT Austin 2014 iGEM [http://2014.igem.org/Team:Austin_Texas/photocage Photocage project], where the part successfully incorporated ONBY within T7 RNA polymerase (RNAP), yielding a light-activated T7 RNAP.

IodoY-UT Austin 2014.png

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 1153
    Illegal BamHI site found at 1159
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 323
    Illegal NgoMIV site found at 1185
    Illegal NgoMIV site found at 1645
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 316
    Illegal SapI.rc site found at 1109