Coding
Part:BBa_K1962006:Design
Designed by: Frank Sargent Group: iGEM16_Dundee (2016-10-11)
Truncated Colicin E9 Lacking Bacteriocin Active Domain
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1381
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1375
Illegal BamHI site found at 1357 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
Compliant with RFC[10].
Source
We used the Chimera molecular modelling software to determine the position of the DNase domain of Colicin E9, we then designed a truncated Colicin E9, without this domain. The protein sequence was then back translated into DNA and codon optimised for E. coli K-12 before being synthesised by IDT as a gBlock gene fragment. Using oligonucleotide primers we then amplified the truncated colicin E9 with a multiple cloning site containing the following restriction sites: BamHI, KpnI, SalI, BglII and NheI at the 3' end.