Composite

Part:BBa_K2443033

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


MK optimized for expression in E. coli

myokinase (MK) is a phosphotransferase enzyme that is responsible for the catalyzing the interconversion of adenine nucleotides (ATP, ADP, and AMP). By constantly monitoring phosphate nucleotide levels inside the cell, ADK plays an important role in cellular energy homeostasis. MK has been Codon optimized for use in Escherichia coli and contains a N-terminal hexahistidine tag. Under the regulation of T7 Promoter, RBS and double terminator. The expected size of MK is 25 kDa, and K2443033 expression is seen in lanes 1 and 2. Protein overexpression was performed in BL21DE3 gold containing pUC57-kan with K2443033 coding sequence inserted. Proteins were expressed according to the overexpression protocol 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_oct_5_gel_4.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 726
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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