Part:BBa_K3792009

NodC Generator I

The NodC Control Generator with Weak Promoter produces the NodC protein. The NodC protein produces the carbohydrate, chitin.The NodC Generator with Weak Promoter does not produce a lot of the NodC protein. The NodC protein produces chitin.

This part consists of 821 base-pair length. It includes a promoter, 1 ribosome binding site, 1 protein-coding sequence, and a terminator. We used promoter K2832184, a weak promoter from the BIOFAB collection. In addition, we also used a protein-coding sequence from Chitin synthase from Rhiozobium leguminosarum.

This generator was assembled into the pSB1K3 plasmid, and then transformed into E.Coli NEB 5-Alpha. To test if the cells were secreting chitin, The FSU 2021 team used the Chitosan Assay Kit (XAN-5126) available from Cell Biolabs. The manual for this kit contains the protocols for converting chitin to chitosan, along with a protocol for detecting chitosan.

Chitin from the cells was converted into chitosan by reacting with 12.5M NaOH at 95°C for 24 hours. The solution was then neutralized with acetic acid. The assay was then performed on the supernatant of the cell culture.

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 AgeI site found at 492
  
• 1000
  COMPATIBLE WITH RFC[1000]