Part:BBa_K4044001:Design
BphP1 (E.coli optimized)
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 760
Illegal PstI site found at 1057
Illegal PstI site found at 1504 - 12INCOMPATIBLE WITH RFC[12]Unknown
- 21INCOMPATIBLE WITH RFC[21]Unknown
- 23INCOMPATIBLE WITH RFC[23]Unknown
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 760
Illegal PstI site found at 1057
Illegal PstI site found at 1504
Illegal NgoMIV site found at 402
Illegal NgoMIV site found at 1153
Illegal NgoMIV site found at 1650 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
Since the BphP1 nucleotide sequence is too large, the two parts of the sequences were obtained separately. Application of TIIS restriction enzymes allow to divide the BphP1 nucleotide sequence into the desired fragments and subsequently to join them. Moreover in our system we needed to fuse additional coding sequences after BphP1, so the given sequense lack its stop codon. To add it into your design in frames of PhytoBrick you will need special linker BBa_K4044020.
Codon optimization of the nucleotide sequence for efficient gene expression in E. coli was performed using GENEWIZ codon optimization tool.
Source
BphP1 protein sequence was taken from https://www.uniprot.org/uniprot/A0A161I5N6 and codon optimized for E. coli.
Source organism: Rhodopseudomonas palustris