Coding

Part:BBa_K5181001:Design

Designed by: Mayukh Mitra   Group: iGEM24_IISER-TVM   (2024-08-08)


P450_truncated: Santalum album


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1162
    Illegal PstI site found at 604
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1162
    Illegal PstI site found at 604
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1162
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1162
    Illegal PstI site found at 604
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1162
    Illegal PstI site found at 604
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Following is the amino acid sequence of the part: MRQKRNNLPPGPPALPIIGNIHILGTLPHQSLYNLAKKYGPIMSMRLGLVPAVVISSPEAAELVLKTHDIVFASRPRLQVADYFHYGTKGVILTEYGTYWRN MRRLCTVKLLNTVKIDSFAGTRKKEVASFVQSLKEASVAHKMVNLSARVANVIENMVCLMVIGRSSDERFKLKEVIQEAAQLAGAFNIGDYVPFLMPLDLQG LTRRIKSGSKAFDDILEVIIDEHVQDIKDHDDEQHGDFIDVLLAMMNKPMDSREGLSIIDRTNIKAILVDMIGAAMDTSTSGVEWAISELIKHPRVMKKLQD EVKTVIGMNRMVEEADLPKLPYLDMVVKETMRLHPPGPLLVPRESMEDITINGYYIPKKSRIIVNAWAIGRDTNAWSNNAHEFFPERFMSSNVDLQGQDFQL IPFGSGRRGCPGMRLGLTTVRLVLAQLIHCFDLELPKGTVATDLDMSEKFGLAMPRAQHLLAFPTYRLESHHHHHH

It has a 6X his-tag at its end and can be purified by a Ni-NTA column.

Removing 64 nucleotide sequences from position 3 to position 67 to remove the transmembrane domain of the protein.


Source

The mother part of this protein is BBa_K5181000, which was taken from the genome of the Santalum album. Then the transmembrane domain was truncated and codon optimized for p.putida

References