Difference between revisions of "Part:BBa K1900001"

 
 
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<partinfo>BBa_K1900001 short</partinfo>
 
<partinfo>BBa_K1900001 short</partinfo>
  
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This part
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Combination of part BBa_K1406000 (BBa_K206000 + BBa_B0034) the E. coli tolC signaling sequence and the Serratia marcescens tolC gene. Transcription is controlled by the arabinose-induced pBAD promoter (BBa_K206000). A strong E. coli RBS (BBa_B0034) controls translation. The signaling sequence is a cleavable N-terminal signaling sequence from E. coli's tolC gene. This is required for trimerization of the TolC monomers as well as insertion into the outer membrane; it forms a chimeric polypeptide with the tolC gene from Serratia marcescens. TolC trimers form a outer-membrane alpha/beta barrel pore can interface with a variety of molecule pumps to form a transperiplasmic pump. <br/> <br/>
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Contribution: <br/>
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Team WLC-Milwaukee 2019 <br/>
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Authors: Laura Sanford, Ben Bateman <br/>
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The tolC gene from Serratia marcescens (part BBa_K1900001) was cloned into a plasmid containing the pTrc promoter. The plasmid was transformed into an E. coli strain that lacked a chromosomal copy of tolC. Disk sensitivity assays were performed to determine how well the S. marcescens protein would work in an E. coli cell. <br/>
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[[File:T--WLC-Milwaukee--BBa_K1900000_graph.JPG]] <br/>
 
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===Usage and Biology===
 
===Usage and Biology===
  
 
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<span class='h3bb'>Sequence and Features</span>
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<span class='h3bb'></span>
 
<partinfo>BBa_K1900001 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1900001 SequenceAndFeatures</partinfo>
  

Latest revision as of 20:05, 21 October 2019


pBAD+strong RBS+E. coli tolC signal sequence+S. marcescens tolC

Combination of part BBa_K1406000 (BBa_K206000 + BBa_B0034) the E. coli tolC signaling sequence and the Serratia marcescens tolC gene. Transcription is controlled by the arabinose-induced pBAD promoter (BBa_K206000). A strong E. coli RBS (BBa_B0034) controls translation. The signaling sequence is a cleavable N-terminal signaling sequence from E. coli's tolC gene. This is required for trimerization of the TolC monomers as well as insertion into the outer membrane; it forms a chimeric polypeptide with the tolC gene from Serratia marcescens. TolC trimers form a outer-membrane alpha/beta barrel pore can interface with a variety of molecule pumps to form a transperiplasmic pump.

Contribution:
Team WLC-Milwaukee 2019
Authors: Laura Sanford, Ben Bateman
The tolC gene from Serratia marcescens (part BBa_K1900001) was cloned into a plasmid containing the pTrc promoter. The plasmid was transformed into an E. coli strain that lacked a chromosomal copy of tolC. Disk sensitivity assays were performed to determine how well the S. marcescens protein would work in an E. coli cell.
T--WLC-Milwaukee--BBa K1900000 graph.JPG


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1642
    Illegal NheI site found at 1663
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1582
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 337
    Illegal NgoMIV site found at 1126
    Illegal NgoMIV site found at 1413
    Illegal AgeI site found at 1311
  • 1000
    COMPATIBLE WITH RFC[1000]