Difference between revisions of "Part:BBa K4060110"
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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.
 
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.
  
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 14:30, 7 December 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.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1112
    Illegal XbaI site found at 1128
    Illegal SpeI site found at 1382
  • 12
    INCOMPATIBLE 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
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1112
    Illegal BamHI site found at 65
    Illegal BamHI site found at 1179
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1112
    Illegal XbaI site found at 1128
    Illegal SpeI site found at 1382
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1112
    Illegal XbaI site found at 1128
    Illegal SpeI site found at 1382
  • 1000
    COMPATIBLE WITH RFC[1000]