Composite

Part:BBa_K2443007

Designed by: Lethbridge iGEM 2017   Group: iGEM17_Lethbridge   (2017-10-18)


GlyRS Alpha subunit optimized for expression in E. coli

Glycine tRNA synthetase responsible for attaching glycine onto its tRNA to form an aminoacyl-tRNA. Codon optimized for use in Escherichia coli. Contains a C-terminal hexahistidine tag with a serine glycine linker. Under the regulation of T7 Promoter, RBS and double terminator. The expected size of GlyRS Alpha subunit is 36 kDa, which closely corresponds to the size observed from the overexpression (lane 5 below). Protein was expressed from BL21DE3 gold from a pUC57-kan plasmid according to the overexpression protocol and notebook on the lethbridge iGEM 2017 wiki (https://static.igem.org/mediawiki/2017/2/2c/Protein_Overexpression_.pdf and https://static.igem.org/mediawiki/2017/0/0d/October_.pdf)

Lethbridge_2017_Oct_10_gel_2.png

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 370
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
//collections/cell-free_tx-tl
Parameters
None