Difference between revisions of "Part:BBa K2014000"
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<b>pBAD-E15'UTR->sfGFP</b> construct is derived from pBAD (Arashort1, BBa_K1741000) Escherichia coli K-12 arabinose promoter, which we fused with <b>E1_5’UTR</b> (Fig. 1). <b>E1_5’UTR</b> contains an additional ribosome binding site from gene 10 of bacteriophage T7. The new promoter controls the expression of sfGFP with a His-tag at its N-end. The fluorescent protein sfGFP is a marker of gene expression and protein synthesis/accumulation. Protein expression from all compared arabinose responsive promoters was induced in rich media with 0.4% L-arabinose.   
 
<b>pBAD-E15'UTR->sfGFP</b> construct is derived from pBAD (Arashort1, BBa_K1741000) Escherichia coli K-12 arabinose promoter, which we fused with <b>E1_5’UTR</b> (Fig. 1). <b>E1_5’UTR</b> contains an additional ribosome binding site from gene 10 of bacteriophage T7. The new promoter controls the expression of sfGFP with a His-tag at its N-end. The fluorescent protein sfGFP is a marker of gene expression and protein synthesis/accumulation. Protein expression from all compared arabinose responsive promoters was induced in rich media with 0.4% L-arabinose.   
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[[Image:https://static.igem.org/mediawiki/parts/8/80/BBa_K2014000-1.png|thumb|400px|alt=Fig. 1 Synthetic evolution of E.coli arabinose induced promoters in our lab.|<font size="2"><b>Fig. 1 Synthetic evolution of E.coli arabinose induced promoters in our lab.</b> pBAD-E15'UTR (Arashort1-E1) is a promoter without AraC ORF, with E1_5’UTR containing the additional ribosome binding site from gene 10 of bacteriophage T7.</font>]]
  
 
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Revision as of 19:00, 19 October 2016

pBAD-E15'UTR->sfGFP

pBAD-E15'UTR->sfGFP construct is derived from pBAD (Arashort1, BBa_K1741000) Escherichia coli K-12 arabinose promoter, which we fused with E1_5’UTR (Fig. 1). E1_5’UTR contains an additional ribosome binding site from gene 10 of bacteriophage T7. The new promoter controls the expression of sfGFP with a His-tag at its N-end. The fluorescent protein sfGFP is a marker of gene expression and protein synthesis/accumulation. Protein expression from all compared arabinose responsive promoters was induced in rich media with 0.4% L-arabinose.

File:Https://static.igem.org/mediawiki/parts/8/80/BBa K2014000-1.png
Fig. 1 Synthetic evolution of E.coli arabinose induced promoters in our lab. pBAD-E15'UTR (Arashort1-E1) is a promoter without AraC ORF, with E1_5’UTR containing the additional ribosome binding site from gene 10 of bacteriophage T7.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 236
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 71
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 53
    Illegal SapI.rc site found at 365