Composite

Part:BBa_K2443003

Designed by: Lethbridge iGEM 2017   Group: iGEM17_Lethbridge   (2017-10-04)


AspRS optimized for expression in E. coli

Aspartic acid tRNA synthetase responsible for attaching aspartic acid 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. AspRS has an expected size of 67 kDa, which corresponds to the band size seen in lanes 2 and 3 below. Protein was overexpressed from BL21DE3 gold where the construct was in pSB1C3. Overexpression was performed according to the overexpression protocols, and 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).

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 BglII site found at 835
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 752


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