Difference between revisions of "Part:BBa K1725351"
Line 1: | Line 1: | ||
− | |||
__NOTOC__ | __NOTOC__ | ||
<partinfo>BBa_K1725351 short</partinfo> | <partinfo>BBa_K1725351 short</partinfo> | ||
− | . | + | Part K1725351 is a <i>luxCDE</i> gene assembly under the R00N1 (<partinfo>K1725080</partinfo>) promoter generated by BioBrick assembly by combining <i>luxC</i> (<partinfo>K1725202</partinfo>), <i>luxD</i> (<partinfo>K1725203</partinfo>) and <i>luxE</i> (<partinfo>K1725204</partinfo>) ribosome binding site (RBS) libraries together. RBS <partinfo>K1725307</partinfo>, <partinfo>K1725302</partinfo> and <partinfo>K1725301</partinfo> are upstream of <i>luxC</i>, <i>luxD</i> and <i>luxE</i> genes, respectively. The assembly has been shown to induce high levels of bioluminescence in <i>E. coli</i> when co-tranformed with pBAD.<i>luxABG</i>-bright (<partinfo>K1725350</partinfo>). [http://2015.igem.org/Team:Glasgow/Description More information] about the part. |
+ | |||
+ | ==Design of RBS library for <i>luxCDE</i>== | ||
+ | For the construction of the RBS library, a master sequence based on the RBS B0032 was used. 4 nucleotides within the actual ribosome binding site were randomised giving 32 different B0032-derived RBS variants. The predicted efficiency of each RBS library member was estimated using RBS Library Calculator. Originally, RBS Library was designed for every gene in luxABGCDE operon (<partinfo>K1725352</partinfo>) in order to optimise bioluminescence in E. coli. 32 different BioBrick RBS variants were incorporated upstream of each of the <i>lux</i> genes by PCR, and different libraries were then combined together by BioBrick assembly. This method could potentially produce over 1 billion different variants of the <i>lux</i> operon. Just for the <i>luxCDE</i> assembly, there are over 32000 different variants. | ||
+ | [[File:Igem_logo_luxCDE.jpg|280px|thumb|upright|iGEM logo painted with <i>E. coli</i> co-transformed with <partinfo>K1725350</partinfo> and <partinfo>K1725351</partinfo>]] | ||
+ | [[File:GlasgowTeam_RBSlibrarydesign.jpg|620px|thumb|none|Design of the RBS library for the <i>lux</i> operon ]] | ||
<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here |
Revision as of 23:01, 18 September 2015
K1725080.luxCDE - bright
Part K1725351 is a luxCDE gene assembly under the R00N1 (BBa_K1725080) promoter generated by BioBrick assembly by combining luxC (BBa_K1725202), luxD (BBa_K1725203) and luxE (BBa_K1725204) ribosome binding site (RBS) libraries together. RBS BBa_K1725307, BBa_K1725302 and BBa_K1725301 are upstream of luxC, luxD and luxE genes, respectively. The assembly has been shown to induce high levels of bioluminescence in E. coli when co-tranformed with pBAD.luxABG-bright (BBa_K1725350). [http://2015.igem.org/Team:Glasgow/Description More information] about the part.
Design of RBS library for luxCDE
For the construction of the RBS library, a master sequence based on the RBS B0032 was used. 4 nucleotides within the actual ribosome binding site were randomised giving 32 different B0032-derived RBS variants. The predicted efficiency of each RBS library member was estimated using RBS Library Calculator. Originally, RBS Library was designed for every gene in luxABGCDE operon (BBa_K1725352) in order to optimise bioluminescence in E. coli. 32 different BioBrick RBS variants were incorporated upstream of each of the lux genes by PCR, and different libraries were then combined together by BioBrick assembly. This method could potentially produce over 1 billion different variants of the lux operon. Just for the luxCDE assembly, there are over 32000 different variants.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 2050
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1435
Illegal SapI.rc site found at 2603