Difference between revisions of "Part:BBa K4417019"
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<h1>Description</h1> | <h1>Description</h1> | ||
| − | 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. | + | This part was created by cloning the master plasmid (<partinfo>BBa_K4417017</partinfo>) with the coding sequence of ureABC (<partinfo>BBa_K4417012</partinfo>) and ureEFG (<partinfo>BBa_K4417013</partinfo>). Both ureABC and ureEFG genes have their own cumate inducible promoter, RBS, and terminator. |
| − | [[File:Lllll.png| | + | [[File:Lllll.png|400px|thumb|center|'''Figure 1:'''CuO-RBS-ureABC-rrnB T1 Terminator-CuO-RBS-ureEFG-rrnB T1 Terminator MTU.]] |
<h1>Usage and Biology</h1> | <h1>Usage and Biology</h1> | ||
| − | * This part can be transformed in both B. subtilis and E. coli. | + | * This part can be transformed in both ''B. subtilis'' and ''E. coli''. |
* Inducer: p-isopropyl benzoate (cumate). | * Inducer: p-isopropyl benzoate (cumate). | ||
* Cumate is non-toxic to the host. In our experiment, we induced the urease expression with 50 μM 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 | + | * ''E. coli'' ori is a pMB1 derivative |
| − | * B. subtilis ori is unknown | + | * ''B. subtilis'' ori is unknown |
| − | * The copy number of this plasmid in B. subtilis and E. coli 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. | * 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. | ||
Latest revision as of 15:48, 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.
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.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE 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 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]