Coding

Part:BBa_K2382004:Design

Designed by: SHAO-CHI LO   Group: iGEM17_CSMU_NCHU_Taiwan   (2017-10-26)


Thioredoxin with polylinker

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 367
    Illegal XhoI site found at 373
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Thioredoxin could help the protein folding correctly, and make the fusion proteins in soluble form that are biologically active.

And we remove the stop coden of Thioredoxin(NC_000913.3) so that it can be used in fusion protein designing.

We Insert linker between our two proteins: thioredoxine and FGD. The purpose to do so is to separate these two proteins.

The sequence of Linker contain the polylinker which has the sequence of GGTACCCGGGGATCCCTCGAGGGTGGT.

The polylinker contain four restriction sites in it:

Kpn1:GGT’ACC

Sma1: CCC’GGG

BamH1:GGA’TCC

Xho1:C’TCGAG

Two glycines are added at the end of the sequence, and it allows the protein behind linker can be reversed, reducing the probability of protein folding error.

Source

We have it synthesized by AllBio Science Incorporated.