Part:BBa_K2848001

NodC+COD-HisTag

This Part allows for the production of chitosan in a single cell with the need of only one plasmid to be transformed. We combined two parts from the iGEM Team Darmstadt 2017 (ChiTUCare), NodC (chitin synthase, BBa_2380002) and COD (chitin deacetylase, BBa_2380043). The COD was improved by adding a His-Tag for easy purification after expression in E.coli BL21 DE3. The use of the BL21 DE3 strain is important, since the COD-ORF is under the control of an IPTG inducible T7 promotor.

[File:T--Stuttgart--Bba_K2848001_Map.png|200px|thumb|left|Plasmid map was created with Snapgene.]

By using two different induction systems, Arabinose for NodC and IPTG for COD, a sequential expression of the emzymes is possible. This might optimize the production of chitosan wich has a very low solubility in aqueos solutions.

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 1144
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 979
  Illegal AgeI site found at 1613
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI site found at 961