Coding
Part:BBa_K2043001:Design
Designed by: Sebastian Sosa-Carrillo and Alicia Calvo-Villamanan Group: iGEM16_Paris_Bettencourt (2016-10-13)
catA from Acinetobacter pittii, codon optimized for E. coli
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
His-tag is added in the C-terminal for protein purification
The sequence was codon optimized for E. coli and restriction sites for the restriction enzymes BpiI, BsaI and BsmBI were avoided.
The sequence was confirmed by sequencing and no mutations were observed.
The construction was done by Golden Gate technique using BsaI as restriction enzyme to obtain the desired cohesive ends.
Source
Acinetobacter pittii's genome, NCBI Reference Sequence for the gene: YP_004995593.1
References
Lin, J., & Milase, R. N. (2015). Purification and Characterization of Catechol 1, 2-Dioxygenase from Acinetobacter sp. Y64 Strain and Escherichia coli Transformants. The protein journal, 34(6), 421-433.