Coding
PheRS

Part:BBa_K2916007

Designed by: Konstantinos Ragios   Group: iGEM19_EPFL   (2019-07-24)


PheRS protein equipped with a 6x HIS affinity tag


Phenylalanyl-tRNA synthetase is a protein that is used in the OnePot PURE cell-free system.


Biology

Transfer RNA (tRNA) is the molecule that enables the Genetic Code contained in the nucleotide sequence of a messenger RNA (mRNA) molecule to be translated into the amino acid sequence of a polypeptide chain. This function is catalysed by a group of enzymes called aminoacyl-tRNA synthetase (aaRS) which attach the appropriate amino acid onto its tRNA. This group consists of 20 different types of aminoacyl-tRNA synthetases, one for each amino acid of the genetic code. Those enzymes, in presence of Adenosine triphosphate (ATP) and Amino Acids, produces aminoacyl-tRNA (tRNA charged with an Amino Acid) that can be used by the ribosome to transfer the amino acid from the tRNA to the polypeptide being synthesised, according of course to the genetic code. Aminoacyl-tRNA therefore play a major role in RNA translation.


Phenylalanyl-tRNA synthetase (PheRS) attaches to Phenylalanine (codons: UUU and UUC)

In our part, besides the sequence encoding for the protein we also have a hexahistidine-tag to allow us purify the protein.


Usage

In our project we used PheRS as a part of the protein solution needed for OnePot PURE cell-free system using the method of gravity flow affinity chromatography, as described in the protocol we designed.



Characterization

All the data regarding the characterization of that part can be found in the composite part: BBa_K2916027 .




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 2424
    Illegal BamHI site found at 31
    Illegal BamHI site found at 1772
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1162
    Illegal AgeI site found at 1155
    Illegal AgeI site found at 2077
    Illegal AgeI site found at 2929
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2875
    Illegal SapI.rc site found at 198
    Illegal SapI.rc site found at 2851


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