Difference between revisions of "Part:BBa K2016000"
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<partinfo>BBa_K2016000 short</partinfo> | <partinfo>BBa_K2016000 short</partinfo> | ||
| − | BBa_K2016000 is a derivative of Bba_J23100, a promoter from a family of constitutive <i>E. coli</i> 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 <i>E. coli</i>, followed by a 50 nucleotide spacer (5' UTR) and ending with a 6 nucleotide ribosomal binding site (AGGAGG).< | + | <html><center><img style="width:35%;float:right;margin: 0px 20px;border:6px double black" src="https://static.igem.org/mediawiki/2016/e/e9/Strp2.png"</center><p align="justify">BBa_K2016000 is a derivative of Bba_J23100, a promoter from a family of constitutive <i>E. coli</i> 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 <i>E. coli</i>, followed by a 50 nucleotide spacer (5' UTR) and ending with a 6 nucleotide ribosomal binding site (AGGAGG).<br/></p></html> |
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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. 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> | + | <html><p align="justify">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). <a href="http://2016.igem.org/Team:Sheffield/project/science/furreporter">Read more about our experiments.</a><br/><br/><center><img style="border:6px double black" src="https://static.igem.org/mediawiki/2016/2/23/Gfpgraph.png"></center></p></html> |
Revision as of 10:02, 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).
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). Read more about our experiments.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 38