Coding

Part:BBa_K1897000:Design

Designed by: Choi Yan Ru   Group: iGEM16_NUS_Singapore   (2016-10-07)
Revision as of 18:48, 10 October 2016 by Cyr95 (Talk | contribs) (→‎References)

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HasA hemophore coding sequence


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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 442


Design Notes

We have included a hexa-histidine tag at the end of the coding sequence as to facilitate protein purification using Immobilised Metal Affinity Chromatography. However, this is only the coding sequence and therefore a relevant promoter, ribosome binding site and terminator would be required for expression (see BBa_K1897001 for the HasA construct that is ready for expression). This construct can be used by others who wish to change the control of the expression of this gene by adding their own promoters.

Source

The HasA gene is originally found in the Serratia marcescens. We synthesised the part from the Serratia marcescens genome, obtaining the gene sequence from the National Center for Biotechnology Information (NCBI) database (http://www.ncbi.nlm.nih.gov/nuccore/X81195.2), including a hexa histadine tag before the stop codon.

References

Serratia marcescens hasA, has B, hasD, hasE, hasI, hasR and hasS genes. Retrieved May 2016 from http://www.ncbi.nlm.nih.gov/nuccore/X81195.2

Cescau, S., Cwerman, H., Letoffe, S., Delepelaire, P., Wandersman, C., & Biville, F. (2007). Heme acquisition by hemophores. Biometals, 20(3-4), 603-613.