Composite

Part:BBa_K2974500:Design

Designed by: Janet Standeven, Abby Bell, Ashtyn Cauffman, Monica Cho   Group: iGEM19_Lambert_GA   (2019-10-16)
Revision as of 18:03, 21 October 2019 by Monicacho123 (Talk | contribs)


Medium Promoter (BBa_J23106) Toehold LacZ


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


Design Notes

The toehold was designed with promoter BBa_J23106 and reporter gene BBa_K2550201. This reporter contributed by 2018 Lambert iGEM codes for the full-length LacZ Beta-galactosidase gene with a wobble introduced to ensure that the gene is restriction fragment cloning compatible. In our model, LacZ is the downstream reporter translated following binding of the trigger sequence to the toehold switch. The entire LacZ construct was designed in SnapGene. Due to the size of the LacZ reporter gene and constraints of restriction sites present in the pSB3C5 plasmid, the team decided to use Gibson Assembly rather than restriction digest in cloning workflow. Gibson primers were designed using SnapGene for the J23106 promoter T7 LacZ Toehold.

T--Lambert GA--Snapgene3.png

Figure 1. Toehold LacZ Construct with BBa_J23106 promoter designed in SnapGene.



T--Lambert GA--Snapgene4.png

Figure 2. LacZ Toehold Construct with BBa_J23100 Promoter designed in SnapGene.

Source

Genomic

References