Difference between revisions of "Part:BBa K4060110"
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<partinfo>BBa_K4060110 short</partinfo> | <partinfo>BBa_K4060110 short</partinfo> | ||
− | a | + | Since bioparts may behave differently in different host strains, any team who wish to integrate this kill switch system into their designs can use Construct 2 (BBa_K4060111) to roughly estimate MazE expression level in various condition. |
+ | After that, a constitutive promoter with appropriate strength could be selected to construct a MazF constant expression system. Strength of constitutive promoters can be test out with an RFP reporter system, and we found that BBa_J23100 constitutive promoter family on BBa_J61002 reporter plasmid is useful for this purpose. A promoter that shows a slightly higher level of RFP expression in comparison with that of Construct 2 (BBa_K4060111) under L-arabinose induction (1mM) or low temperature (20℃) should be chosen. Integrate Construct 1 (BBa_K4060110) with the aforementioned MazF constant expression system in the same plasmid, and the kill switch system is completed. | ||
+ | |||
+ | <div class="img"> | ||
+ | <img src="https://2021.igem.org/wiki/images/d/df/T--NYCU-Taipei--const1const2.png" alt=""> | ||
+ | </div> | ||
<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here |
Revision as of 03:55, 22 October 2021
Const1
Since bioparts may behave differently in different host strains, any team who wish to integrate this kill switch system into their designs can use Construct 2 (BBa_K4060111) to roughly estimate MazE expression level in various condition. After that, a constitutive promoter with appropriate strength could be selected to construct a MazF constant expression system. Strength of constitutive promoters can be test out with an RFP reporter system, and we found that BBa_J23100 constitutive promoter family on BBa_J61002 reporter plasmid is useful for this purpose. A promoter that shows a slightly higher level of RFP expression in comparison with that of Construct 2 (BBa_K4060111) under L-arabinose induction (1mM) or low temperature (20℃) should be chosen. Integrate Construct 1 (BBa_K4060110) with the aforementioned MazF constant expression system in the same plasmid, and the kill switch system is completed.
<img src="" alt="">
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 1112
Illegal XbaI site found at 1128
Illegal SpeI site found at 1382 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 1112
Illegal NheI site found at 125
Illegal SpeI site found at 1382
Illegal NotI site found at 1119 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 1112
Illegal BamHI site found at 65
Illegal BamHI site found at 1179 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 1112
Illegal XbaI site found at 1128
Illegal SpeI site found at 1382 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 1112
Illegal XbaI site found at 1128
Illegal SpeI site found at 1382 - 1000COMPATIBLE WITH RFC[1000]