Difference between revisions of "Part:BBa K5133001"

Revision as of 05:56, 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 sfGFP expression for cell-free protein synthesis (CFPS). 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 demonstrate the correctness of DNA sequence, result of Sanger sequencing for BBa_K5133004 show the successful assembly among T7 promoter (BBa_K5133001), 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

Sequence and Features