Difference between revisions of "Part:BBa K5133001"
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==<b>Brief introduction</b>== | ==<b>Brief introduction</b>== | ||
| − | This basic part is derived from plasmid pJL1 (Addgene: #69496)<sup>[1]</sup>, including a conserved ribosome binding site (RBS) as <b>5'-aagaaggaga-3'</b><sup>[2]</sup>. The plasmid pJL1 is commonly used for the <i>in vitro</i> sfGFP expression of cell-free protein synthesis (CFPS). Hence, this part is used for the construction of three composite parts: <bbpart>BBa_K5133004</bbpart> (sfGFP generator), <bbpart>BBa_K5133006</bbpart> (Microcin H47 generator), and <bbpart>BBa_K5133008</bbpart> (Microcin M generator), for CFPS in our project. | + | This basic part is derived from plasmid pJL1 (Addgene: #69496)<sup>[1]</sup>, including a conserved ribosome binding site (RBS) as <b>5'-aagaaggaga-3'</b><sup>[2]</sup>. The plasmid pJL1 is commonly used for the <i>in vitro</i> sfGFP expression of cell-free protein synthesis (CFPS)<sup>[3]</sup>. Hence, this part is used for the construction of three composite parts: <bbpart>BBa_K5133004</bbpart> (sfGFP generator), <bbpart>BBa_K5133006</bbpart> (Microcin H47 generator), and <bbpart>BBa_K5133008</bbpart> (Microcin M generator), for CFPS in our project. |
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[2] Hausjell, J. et al. The effects of lactose induction on a plasmid-free <i>E. coli</i> T7 expression system. <b>Bioengineering</b> 7, 8 (2020). doi: 10.3390/bioengineering7010008 | [2] Hausjell, J. et al. The effects of lactose induction on a plasmid-free <i>E. coli</i> T7 expression system. <b>Bioengineering</b> 7, 8 (2020). doi: 10.3390/bioengineering7010008 | ||
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| + | [3] Ba, F. et al. Expanding the toolbox of probiotic <i>Escherichia coli</i> Nissle 1917 for synthetic biology. <b>Biotechnology Journal</b> 19, 2300327 (2024)). doi: 10.1002/biot.202300327 | ||
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Revision as of 06:16, 27 July 2024
Ribosome binding site (RBS, from plasmid pJL1)
Group: GEC-China (iGEM 2024, team number: #5133)
Brief introduction
This basic part is derived from plasmid pJL1 (Addgene: #69496)[1], including a conserved ribosome binding site (RBS) as 5'-aagaaggaga-3'[2]. The plasmid pJL1 is commonly used for the in vitro sfGFP expression of cell-free protein synthesis (CFPS)[3]. Hence, this part is used for the construction of three composite parts: BBa_K5133004 (sfGFP generator), BBa_K5133006 (Microcin H47 generator), and BBa_K5133008 (Microcin M generator), for CFPS in our project.
Design and characterization
The plasmid design of this biological part is shown as Figure 1, assembled with iGEM standard backbone pSB1C3. To validate the correctness of DNA sequence, result of Sanger sequencing for BBa_K5133004 show the successful assembly among T7 promoter (BBa_K5133000), RBS (this part), and sfGFP (BBa_K5133002) (Figure 2).
Usages
This part is used for the construction of three composite parts: BBa_K5133004 (sfGFP generator), BBa_K5133006 (Microcin H47 generator), and BBa_K5133008 (Microcin M generator), for CFPS in our project.
DNA sequence (from 5' to 3')
aagaaggagatatacat
Red font: RBS
References
[1] https://www.addgene.org/69496/
[2] Hausjell, J. et al. The effects of lactose induction on a plasmid-free E. coli T7 expression system. Bioengineering 7, 8 (2020). doi: 10.3390/bioengineering7010008
[3] Ba, F. et al. Expanding the toolbox of probiotic Escherichia coli Nissle 1917 for synthetic biology. Biotechnology Journal 19, 2300327 (2024)). doi: 10.1002/biot.202300327
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]