Difference between revisions of "Part:BBa K4417019"

 
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<partinfo>BBa_K4417019 short</partinfo>
 
<partinfo>BBa_K4417019 short</partinfo>
  
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<h1>Description</h1>
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This part was created by cloning the master plasmid (BBa_K4417017) with the coding sequence of ureABC (BBa_K4417012) and ureEFG (BBa_K4417013). Both ureABC and ureEFG genes have their own cumate inducible promoter, RBS, and terminator.
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[[File:Lllll.png|600px|thumb|center|'''Figure 1:'''CuO-RBS-ureABC-rrnB T1 Terminator-CuO-RBS-ureEFG-rrnB T1 Terminator MTU.]]
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<h1>Usage and Biology</h1>
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* This part can be transformed in both B. subtilis and E. coli.
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* Inducer: p-isopropyl benzoate (cumate).
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* Cumate is non-toxic to the host. In our experiment, we induced the urease expression with 50 μM cumate.
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* E. coli ori is a pMB1 derivative
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* B. subtilis ori is unknown
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* The copy number of this plasmid in B. subtilis and E. coli is unknown.
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* NdeI restriction can be used to change promoter. In this way, the performance of ureABC and ureEFG could be compared, each from a different promoter.
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<h1>Method</h1>
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Golden Gate Assembly was used to assemble the master plasmid with TU1 and TU2.
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[[File:Uhuhuhu.png|600px|thumb|center]]
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Revision as of 15:12, 12 October 2022


CuO-RBS-ureABC-rrnB T1 Terminator-CuO-RBS-ureEFG-rrnB T1 Terminator MTU

Description

This part was created by cloning the master plasmid (BBa_K4417017) with the coding sequence of ureABC (BBa_K4417012) and ureEFG (BBa_K4417013). Both ureABC and ureEFG genes have their own cumate inducible promoter, RBS, and terminator.

Figure 1:CuO-RBS-ureABC-rrnB T1 Terminator-CuO-RBS-ureEFG-rrnB T1 Terminator MTU.


Usage and Biology

  • This part can be transformed in both B. subtilis and E. coli.
  • Inducer: p-isopropyl benzoate (cumate).
  • Cumate is non-toxic to the host. In our experiment, we induced the urease expression with 50 μM cumate.
  • E. coli ori is a pMB1 derivative
  • B. subtilis ori is unknown
  • The copy number of this plasmid in B. subtilis and E. coli is unknown.
  • NdeI restriction can be used to change promoter. In this way, the performance of ureABC and ureEFG could be compared, each from a different promoter.

Method

Golden Gate Assembly was used to assemble the master plasmid with TU1 and TU2.

Uhuhuhu.png


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 3844
    Illegal BamHI site found at 1
    Illegal BamHI site found at 2779
    Illegal XhoI site found at 4598
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]