Coding

Part:BBa_K4044001:Design

Designed by: Kalinichenko Andrei   Group: iGEM21_LMSU   (2021-10-19)
Revision as of 21:49, 21 October 2021 by Andrei (Talk | contribs) (adding about stop codon)


BphP1 (E.coli optimized)


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 760
    Illegal PstI site found at 1057
    Illegal PstI site found at 1504
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 760
    Illegal PstI site found at 1057
    Illegal PstI site found at 1504
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 246
    Illegal BamHI site found at 1821
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 760
    Illegal PstI site found at 1057
    Illegal PstI site found at 1504
  • 25
    INCOMPATIBLE 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
  • 1000
    COMPATIBLE 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

References