Part:BBa_K2841515
MG1655 Specific EndA Mutant Recognition Device
The targeting region of a dCAS9 (BBa_K2841510) gRNA (BBa_K2841514) will become exposed in response to the E.Coli K12 MG1655 mutant of the EndA gene, but not the E.Coli K12 DH5 Alpha mutant. Once the targeting region of the gRNA is exposed the LacIq promoter (BBa_K2841517) will be down regulated in response. In combination with a reporter circuit, the GFP per OD600 can be used to determine the strain of E.coli that has been transformed. The theoretical part has never been tested, but should work with the system utilised in the Warwick iGEM 2018 design.
The sequence recognised is: TGAGCGCAGCATTTT CCGGCCCGG CGTTGGCCGAAGGTA
The bold region is identified in the loop and the italicised region is identified further upstream in the gRNA for the purpose of specificity.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 158
Illegal NgoMIV site found at 187 - 1000COMPATIBLE WITH RFC[1000]
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