Coding

Part:BBa_K2127001:Design

Designed by: Nafaa Haddou   Group: iGEM16_IngenuityLab_Canada   (2016-10-14)
Revision as of 13:44, 21 October 2016 by Nhaddou (Talk | contribs) (→‎Design Notes)


Inducible High Expression His-Tagged Photosystem II CP47 subunit


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 658
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 782
    Illegal AgeI site found at 1379
    Illegal AgeI site found at 1583
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1103
    Illegal SapI site found at 1780


Design Notes

Cyanobacteria is not optimal system to manipulate the protein expression mainly due of the lack of strong promoters. Pcpc560 is a Cyanobacterial super promoter discovered by Dr. Ma’s team that contains two predicted promoters 14 predicted transcription binding site. Using the Pcpc560, Dr. Ma’s team demonstrated that functional protein were produced at 15% of the total protein production. Our team decided to test the effect protein production in E.Coli cells DH5α using the 14 predicted transcription binding site from Pcpc560. We believe that 14 sequential binding sites from Pcpc560 will allow us to increase the protein function production greatly using the E.Coli DH5α. We have constructed the 14 transcription binding followed by an strong RBS (BBa_B0030), followed by mutant HIS-Tag psbB. The super promoter can be an valuable asset to other iGEM teams for protein production and therefore we have decided to submit the Construct along with 14 Transcription binding site from Pcpc560.

Source

Synchocisytys

References