RBS

Part:BBa_K3192030

Designed by: Simonne Guenette   Group: iGEM19_Virginia   (2019-10-16)


Custom RBS Optimzed for BBa_K3192018

Synthetic Ribosomal Binding site for BBa_K3192018, a BioBrick created for split kanamycin resistance to maintain two plasmids within one cell using a single antibiotic resistance gene.

The 2019 Virginia iGEM Team sought efficiency and productivity in designing their genetic construct as the device required two plasmids to operate. Two plasmids bears a large metabolic load on the chassis, so, synthetic ribosomal binding sites were developed in order to optimize translations rates for the genes used in the styrene-degrading and polyhydroxybutyrate-producing plasmids. This ribosomal binding site is optimized for part BBa_K3192018 with a translation initiation rate of 524834.67.

When expressing gene clusters in a foreign chassis, the endogenous ribosomal binding sites (RBS) are often included in the sequence information provided by NCBI. Although these RBS are sufficient, they may not provide optimal translational efficiency. Synthetic RBSs provide a solution to increase translation initiation rates by using software to design a RBS with a linear secondary structure that allows for optimal ribosomal binding and translation. Our team’s project required enhanced translation of our coding sequences; therefore we chose to optimize all RBSs for high translation initiation rates. Using DeNovo DNA, our team created synthetic RBSs that ran thousands of iterations to create substantially elevated translation initiation rates.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 4
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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