Difference between revisions of "Part:BBa K4137013"

 
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This part consists of a pLac promoter, RBS, a ccdB-6xHis coding complex, and a terminator (BBa_B1006). This part produces a constant expression of ccdB toxin under the LB medium, inhibiting the GyrA subunit of DNA gyrase and prevents negative supercoiling of DNA. An accumulation of ccdB without sufficient suppression by antitoxin ccdA leads to risks in host viability.  
 
This part consists of a pLac promoter, RBS, a ccdB-6xHis coding complex, and a terminator (BBa_B1006). This part produces a constant expression of ccdB toxin under the LB medium, inhibiting the GyrA subunit of DNA gyrase and prevents negative supercoiling of DNA. An accumulation of ccdB without sufficient suppression by antitoxin ccdA leads to risks in host viability.  
  
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[[File:plac-ccdb-his.png|800px|thumb|center|Fig.1 Construct design for ccdB toxin secretion.]]
  
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===Construct Design===
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===Usage and Biology===
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We attached a 6x His-Tag downstream of the ccdB sequence for purification purposes. A pLac promoter, which induces protein production when malate is present, is attached upstream to the side of the open reading frame + RBS. The terminator BBa_B1006 is attached downstream of the sequence.
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===Characterization===
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===Sequence and Features===
  
 
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Revision as of 06:24, 9 October 2022


ccdB toxin secretion pathway

This part consists of a pLac promoter, RBS, a ccdB-6xHis coding complex, and a terminator (BBa_B1006). This part produces a constant expression of ccdB toxin under the LB medium, inhibiting the GyrA subunit of DNA gyrase and prevents negative supercoiling of DNA. An accumulation of ccdB without sufficient suppression by antitoxin ccdA leads to risks in host viability.

Fig.1 Construct design for ccdB toxin secretion.

Construct Design

We attached a 6x His-Tag downstream of the ccdB sequence for purification purposes. A pLac promoter, which induces protein production when malate is present, is attached upstream to the side of the open reading frame + RBS. The terminator BBa_B1006 is attached downstream of the sequence.

Characterization

Sequence and Features

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 289