Coding

Part:BBa_K4169029:Design

Designed by: Sijia Xu   Group: iGEM22_HZAU-China   (2022-10-11)
Revision as of 08:09, 12 October 2022 by Lloheha (Talk | contribs) (→‎Source)


Mutated TMADH


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 388
    Illegal PstI site found at 183
    Illegal PstI site found at 1782
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 388
    Illegal PstI site found at 183
    Illegal PstI site found at 1782
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 388
    Illegal XhoI site found at 1717
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 388
    Illegal PstI site found at 183
    Illegal PstI site found at 1782
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 388
    Illegal PstI site found at 183
    Illegal PstI site found at 1782
    Illegal AgeI site found at 879
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

GenScript synthesised tmd plasmids and performed codon optimization. We added His tag on the end of tmd sequence. Then optimized trimethylamine dehydrogenase, mutated amino acid 344 form Val to Cys.The primers we designed for mutation are showned below:

V344C-F: GACGACATCCGTTGTTGTATCGGCTG

V344C-R: CAGCCGATACAACAACGGATGTCGTC

Source

The original gene sequence of tmd is from The European Nucleotide Archive (ENA). Coding: CDX46666.1.

The link is here: https://www.ebi.ac.uk/ena/browser/view/CDX46666

And tmd gene is originally in Mesorhizobium sp. ORS 3359.

References