Part:BBa_K1416001

The tRNA synthetase/tRNA needed for incorporating 3-iodo-L-tyrosine (IodoY) at a UAG codon

The part includes the gene coding for the Methanocaldococcus jannaschii synthetase that charges the orthogonal tRNA (also in the part) with the amino acid 3-iodotyrosine along with the proper promoters and terminator. This part encodes for the incorporation of tyrosine during translation at AUG amber stop codons in a cell with Release Factor 1 (RF1) knocked out such as "Amberless" E. coli or "RF0".

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 this part was one of the best ncAA tRNA synthetase/tRNA pairs

In the control cultures, IodoY-pFRYC, the level of GFP expression relative to RFP expression is set as a standard. In the test cultures, IodoY-pFRY, the level of GFP expression relative to RFP expression indicates how well the part function. In the absence of ncAA, GFP expression is about 20% of RFP expression, a much lower value than in the control conditions. In the presence of ncAA, the amount of GFP expression increases back to roughly 100% of RFP expression. These results indicate that part BBa_K1416001 has good fidelity, and is very efficient. For more information and details, please visit [http://2014.igem.org/Team:Austin_Texas/kit#Results_and_Data ncAA kit project]

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 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