Part:BBa_K5109016:Design
Dehalogenase type 2 variant 2 with His Tag
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 706
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 1
Design Notes
The sequence has two different enzyme restriction sites at the ends: a BsaI restriction site upstream the coding sequence, and a BamHI restriction site downstream the sequence, after the His - Tag. Those sites have been designed in order to extract the dehalogenase sequence from the surface display system in which it is inserted (BBa_K5109023), in order to exchange it with other enzymes, without modifying the rest of the expression cassette. This part was designed for a multiple directed cloning experiment that involved exchanging it with three different enzymes, described in parts BBa_K5109013, BBa_K5109014, and BBa_K5109015.
Source
The original sequence of Dehalogenase type II is found in the genome of Delftia acidovorans. The sequence comes from an improved version of the original sequence, that was modified and optimized for the expression in E. coli.