Composite

Part:BBa_K2443008

Designed by: Lethbridge iGEM 2017   Group: iGEM17_Lethbridge   (2017-10-18)
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GlyRS Beta 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 beta subunit is 78 kDa, and a band of this size can be seen in lane 10 of the overexpression gel below. Protein was expressed from BL21DE3 gold from a pUC57-kan plasmid according to the overexpression protocol and lab notebook on the 2017 iGEM Lethbridge wiki (https://static.igem.org/mediawiki/2017/2/2c/Protein_Overexpression_.pdf and https://static.igem.org/mediawiki/2017/0/0d/October_.pdf)

BBa_K2443001-Oct_17_gel1.png

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1214
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1747
    Illegal SapI site found at 104


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