Difference between revisions of "Part:BBa J100198"

Latest revision as of 19:48, 21 December 2014

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

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal EcoRI site found at 1
Illegal PstI site found at 995
12
INCOMPATIBLE WITH RFC[12]
Illegal EcoRI site found at 1
Illegal PstI site found at 995
21
INCOMPATIBLE WITH RFC[21]
Illegal EcoRI site found at 1
23
INCOMPATIBLE WITH RFC[23]
Illegal EcoRI site found at 1
Illegal PstI site found at 995
25
INCOMPATIBLE 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
1000
COMPATIBLE WITH RFC[1000]

@@ Line 2: / Line 2: @@
 <partinfo>BBa_J100198 short</partinfo>
-Cloned in the fitness gene tester construct ([[Part:BBa_J100199]]), part J100198 contains the fitness gene thymidylate synthase [[Part:BBa_J100132]]. J100198 contains theophylline riboswitch D downstream of T5, a constitutive promoter [[Part:BBa_J100065]]. The construct is built to be used in thymine auxotrophs (''thyA-'') that necessitate 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]]. <br><br>
+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]]. <br><br>
 See Experience page [[Part:BBa_J100198:Experience]] for details about how well this device works.