Generator

Part:BBa_K1499203:Design

Designed by: Raman Nelakanti   Group: iGEM14_StanfordBrownSpelman   (2014-10-09)


SdaB protein generator without tRNA + 5 amber stops


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 6
    Illegal NheI site found at 29
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 226
    Illegal AgeI site found at 549
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 520
    Illegal SapI.rc site found at 667


Design Notes

The SdaB was codon optimized again from the reference sequence. The 5 stop codons were introduced distributed in the protein to secure against expression in organisms other than amberless cells.

The part could not be cloned with the supP tRNA because of toxicity, so the promoter/RBS, protein, and terminator were only included in this part. The tRNA should be either ligated to the end of this part or integrated into the amberless genomic DNA.

Source

SdaB modified from Stanford-Brown's 2012 iGEM part. The part was synthesized using Integrated DNA Technologies and the part minus the tRNA was cloned and put into pSB1C3.

References