Translational_Unit

Part:BBa_K2649012:Design

Designed by: Alexis Schneider   Group: iGEM18_MIT   (2018-10-12)


hEF1a: eComE


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 6
    Illegal EcoRI site found at 1210
    Illegal PstI site found at 337
    Illegal PstI site found at 842
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 6
    Illegal EcoRI site found at 1210
    Illegal PstI site found at 337
    Illegal PstI site found at 842
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 6
    Illegal EcoRI site found at 1210
    Illegal BglII site found at 591
    Illegal BamHI site found at 1197
    Illegal XhoI site found at 990
    Illegal XhoI site found at 1230
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 6
    Illegal EcoRI site found at 1210
    Illegal PstI site found at 337
    Illegal PstI site found at 842
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 6
    Illegal EcoRI site found at 1210
    Illegal PstI site found at 337
    Illegal PstI site found at 842
    Illegal NgoMIV site found at 725
    Illegal AgeI site found at 103
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1227


Design Notes

We chose to drive eComE with hEF1a to have strong constitutive expression.


Source

The eComE protein is from the Streptococcus mutans strain SM6. The protein sequence is item A0A0E2EP61 in the UniProtKB database, and the corresponding codon sequence was optimized for human expression. The NLS, VP16, and NES sequences were taken from Systematic Transfer of Prokaryotic Sensors and Circuits to Mammalian Cells (Weiss and Voigt et al., 2014. DOI: 10.1021/sb5002856)

References