Coding

Part:BBa_K3794000:Design

Designed by: Harshraj Bumia   Group: iGEM21_KCL_UK   (2021-09-30)


PVFP-5


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Removed signal sequence from UniProt U5Y3S6. Codon optimised for expression in E.Coli K12.

Added N-terminal 6xHis tag with start codon and TEV cleavage site for protein purification via Ni-NTA column

Upstream of 6xHis tag, a start codon (ATG) was added to allow for transcriptional initiation, and a stop codon was added at 3' end of PVFP-5 CDS to allow for translational termination during protein synthesis



Source

This part was synthesised as a composite by IDT (with an upstream LacI regulatory (BBa_R0010). The protein sequence was obtained from UniProtKB: U5Y3S6, a 6xHis tag and TEV cleavage site was added, and codon optimised for expression in E.Coli K12

References