Difference between revisions of "Part:BBa K2443007"

Latest revision as of 18:00, 1 November 2017

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)

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]

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 <partinfo>BBa_K2443007 short</partinfo>
-Glycine tRNA synthetase responsible for attaching glycine onto its tRNA to form an aminoacyl-tRNA. Codon optimized for use in <i>Escherichia coli</i>. 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 according to the overexpression protocol on the lethbridge iGEM 2017 wiki (http://2017.igem.org/Team:Lethbridge/Experiments)
+Glycine tRNA synthetase responsible for attaching glycine onto its tRNA to form an aminoacyl-tRNA. Codon optimized for use in <i>Escherichia coli</i>. 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)
 https://static.igem.org/mediawiki/parts/d/d4/Lethbridge_2017_Oct_10_gel_2.png