Coding

Part:BBa_K2770009

Designed by: Jana Anton, Levin Hafa, Leon Schroeder, Sergei Senger   Group: iGEM18_TU_Darmstadt   (2018-09-30)


Alcohol dehydrogenase (adh)


The alcohol dehydrogenase from Lactobacillus kefir is a NADP+-dependent enzyme, which catalyzes the production of aliphatic and aromatic ketones as well as β-keto esters from their respective alcohol. Here we use the ADH to oxidize cyclohexanol to cyclohexanone and as a recycling agent for the cofactor NADP+, which is needed for the CHMO to function. The ADH coding sequence (CDS) is cloned into the pSB1C3 vector, containing the BioBrick prefix and suffix, a T7/lac Promotor (K921001), an RBS (B0034), an N-terminal 10x His-tag for purification and a GS-Linker. The coding sequence consists of 756 bp which are translated to 252 amino acids. ADH has a molecular mass of 26.8 kDa.

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 268
    Illegal AgeI site found at 34
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None