Coding
Part:BBa_K2043005:Design
Designed by: Alicia Calvo-Villamanan y Sebastian Sosa-Carrilo Group: iGEM16_Paris_Bettencourt (2016-10-13)
xylE-FBD1 from Pseudomonas putida codon optimized for E. coli
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 700
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
Fabric Binding Domain 1 was fused downstream in the 5' end of the original catA CDS
A linker was introduced in the DNA sequence between FBD1 and the CDS of catA
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
Pseudomonas putida