Difference between revisions of "Part:BBa K1725351"

Latest revision as of 23:59, 8 February 2017

K1725080.luxCDE - bright

Part K1725351 is a luxCDE gene assembly under the R0011-N (BBa_K1725080) promoter generated by BioBrick assembly of luxC (BBa_K1725202), luxD (BBa_K1725203) and luxE (BBa_K1725204) ribosome binding site (RBS) libraries.

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 the 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.

Error creating thumbnail: Invalid thumbnail parameters

iGEM logo painted with E. coli co-transformed with BBa_K1725350 and BBa_K1725351

Design of the RBS library for the lux operon

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 1
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 2050
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 1435
Illegal SapI.rc site found at 2603

@@ Line 2: / Line 2: @@
 <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.
+Part K1725351 is a <i>luxCDE</i> gene assembly under the R0011-N (<partinfo>K1725080</partinfo>) promoter generated by BioBrick assembly of <i>luxC</i> (<partinfo>K1725202</partinfo>), <i>luxD</i> (<partinfo>K1725203</partinfo>) and <i>luxE</i> (<partinfo>K1725204</partinfo>) ribosome binding site (RBS) libraries.
-==Design of RBS library for <i>luxCDE</i>==
+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 the 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>]]