Composite

Part:BBa_K2797002:Design

Designed by: Frank Eardley   Group: iGEM18_Newcastle   (2018-09-27)
Revision as of 14:21, 2 October 2018 by Frank Eardley (Talk | contribs) (→‎Design Notes)


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 gene 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 (J23119) and RBS (B0034) was added before the aadA sequence. A native terminator (K2797001) was added after the aadA sequence to prevent any read through. The entire construct was flanked by 22 and 21 base pair biobrick prefix and suffixes and was Gibson assembled into a pSB1C3 plasmid digested with XbaI and Spe. The construct was submitted to IDT for synthesis as a gBlock.

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