Difference between revisions of "Part:BBa K5133001"
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==<b>DNA sequence (from 5' to 3')</b>==
 
==<b>DNA sequence (from 5' to 3')</b>==
  
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atcccgcgaaat<font color="red">taatacgactcactatagggaga</font>ccacaacggtttccctctagaaataattttgtttaacttt
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<font color="red">aagaaggaga</font>tatacat
  
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<font color="red">Red font: conserved T7 promoter</font>
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<font color="red">Red font: RBS</font>
  
  
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[1] https://www.addgene.org/69496/
 
[1] https://www.addgene.org/69496/
  
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[2] Conrad, T. et al. Maximizing transcription of nucleic acids with efficient T7 promoters. <b>Communications Biology</b> 3, 439 (2020). doi: 10.1038/s42003-020-01167-x
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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
  
  

Revision as of 05:53, 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, Results of Sanger sequencing for BBa_K5133004 showing the successful assembly among T7 promoter, RBS, and sfGFP (Figure 2).


Resizable Image


Figure 1. Design of basic part BBa_K5133000, generated by SnapGene.



Resizable Image


Figure 2. Validation the DNA sequence of BBa_K5133000 by Sanger sequencing, generated by SnapGene.




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


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]