Difference between revisions of "Part:BBa K1416000"

Revision as of 23:40, 14 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.

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

@@ Line 2: / Line 2: @@
 <partinfo>BBa_K1416000 short</partinfo>
-The part includes the gene coding for the <i>Methanocaldococcus jannaschii</i> synthetase mutated to charge the orthogonal tRNA (also in the part) with the non-canonical amino acid <i>o</i>-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" <i>E. coli</i> or "RF0" <i>E. coli</i>.
+The part includes the gene coding for the <i>Methanocaldococcus jannaschii</i> synthetase mutated to charge the orthogonal tRNA (also in the part) with the non-canonical amino acid <i>o</i>-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" <i>E. coli</i> or "RF0" <i>E. coli</i>.
+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.
 <!-- Add more about the biology of this part here