Composite

Part:BBa_K2797002:Design

Designed by: Frank Eardley   Group: iGEM18_Newcastle   (2018-09-27)
Revision as of 23:32, 17 October 2018 by Frank Eardley (Talk | contribs) (→‎Design Notes)

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Composite part for streptomycin resistance


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal NheI site found at 861
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 709
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The aadA coding sequence was taken from the Addgene database (plasmid #74703). The sequence was found to have no illegal restriction sites (i.e. no EcoRI, XbaI, SpeI, or PstI sites). A strong, standard constitutive promoter (BBa_J23119) and ribosome binding site (BBa_B0034) were added before the aadA sequence. A native terminator (BBa_K2797001) was added after the aadA sequence. The composite part was flanked by 22 and 21 base pair BioBrick prefix and suffixes respectively. The composite part was synthesised as a gBlock by IDT. Gibson assembly was used to clone the gBlock into the pSB1C3 backbone.

Source

Addgene plasmid #74703

References

Genetic circuit design automation. Nielsen AA, Der BS, Shin J, Vaidyanathan P, Paralanov V, Strychalski EA, Ross D, Densmore D, Voigt CA. Science. 2016 Apr 1;352(6281):aac7341. doi: 10.1126/science.aac7341. 10.1126/science.aac7341