Difference between revisions of "Part:BBa K2443008"

Latest revision as of 18:15, 1 November 2017

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)

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

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 <partinfo>BBa_K2443008 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 beta subunit is 78 kDa, and a band of this size can be seen in lane 10 of the overexpression gel below.
+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 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)
 https://static.igem.org/mediawiki/parts/e/e3/BBa_K2443001-Oct_17_gel1.png