Part:BBa_J100198
J100198 contains the fitness gene tester construct (J100199) with thymidylate synthase as the fitnes
Cloned in the fitness gene tester construct (Part:BBa_J100199), part J100198 contains the fitness gene thymidylate synthase Part:BBa_J100132. We used Golden Gate Assembly (GGA) to remove the spacer region from Part:BBa_J100199 and ligate in a wild type thyA allele. J100198 also contains contains the constitutive T5 promoter, followed by theophylline riboswitch D Part:BBa_J100065 followed by the thymidylate synthase Part:BBa_J100132 CDS. The construct is built to be used in thymine auxotrophic E. coli (thyA-) that normally require added thymine in the growth media for cell division. Upon successful binding of theophylline on the riboswitch (we used 0.3mM), thymidylate synthase mRNA is translated for prototophic growth and cellular division in the thyA- chassis. J100198 was cloned into pSB1A8 Part:BBa_J119043.
See Experience page Part:BBa_J100198:Experience for details about how well this device works.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 1
Illegal PstI site found at 995 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 1
Illegal PstI site found at 995 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 1
- 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 1
Illegal PstI site found at 995 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 1
Illegal PstI site found at 995
Illegal AgeI site found at 107
Illegal AgeI site found at 770 - 1000COMPATIBLE WITH RFC[1000]
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