Difference between revisions of "Part:BBa K2016000"

(Usage and Biology)
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===Usage and Biology===
 
===Usage and Biology===
  
Sheffield 2016 has used this promoter in order to design a biological device that responds to intracellular iron levels in bacteria. Bba_J23106 was also used in order to observe a difference between having a system under the control of a medium and a strong promoter.<center>https://static.igem.org/mediawiki/2016/2/23/Gfpgraph.png</center>
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Sheffield 2016 has used this promoter in order to design a biological device that responds to intracellular iron levels in bacteria. Bba_J23106 was also used in order to observe a difference between having a system under the control of a medium and a strong promoter. In the figure below we can see differences in the GFP fluorescence when expressed using a medium promoter (J23106 derivative) or strong promoter (J23100 derivative) in two different strains: W3110 (WT) and JC28 (∆entC)<center>https://static.igem.org/mediawiki/2016/2/23/Gfpgraph.png</center>
  
  

Revision as of 09:05, 18 October 2016


Strong constitutive E. coli promoter with included RBS - ready for cloning

BBa_K2016000 is a derivative of Bba_J23100, a promoter from a family of constitutive E. coli promoters isolated from a small combinatorial library. This family was submitted to the registry by Berkeley 2006. Sheffield 2016 has improved Bba_J23100 by adding an RBS downstream of the promoter to make it suitable for direct cloning. BBa_K2016000 is 91bp long (Fig. 1), first 35 nucleotides make up a functional strong constitutive promoter for expression in E. coli, followed by a 50 nucleotide spacer (5' UTR) and ending with a 6 nucleotide ribosomal binding site (AGGAGG).
Strp2.png


Usage and Biology

Sheffield 2016 has used this promoter in order to design a biological device that responds to intracellular iron levels in bacteria. Bba_J23106 was also used in order to observe a difference between having a system under the control of a medium and a strong promoter. In the figure below we can see differences in the GFP fluorescence when expressed using a medium promoter (J23106 derivative) or strong promoter (J23100 derivative) in two different strains: W3110 (WT) and JC28 (∆entC)
Gfpgraph.png


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 38

Functional Parameters