Difference between revisions of "Part:BBa K4137007"
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<partinfo>BBa_K4137007 short</partinfo> | <partinfo>BBa_K4137007 short</partinfo> | ||
− | + | 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. | |
[[File:plac-ccdb-myc.png|800px|thumb|center|Fig.1 Construct design for ccdB toxin secretion.]] | [[File:plac-ccdb-myc.png|800px|thumb|center|Fig.1 Construct design for ccdB toxin secretion.]] | ||
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===Construct Design=== | ===Construct Design=== | ||
− | We attached a Myc epitope Tag downstream of the ccdB sequence for purification purposes. A pLac promoter is attached upstream to the side of the open reading frame | + | We attached a Myc epitope Tag downstream of the ccdB sequence for purification purposes. A pLac promoter is attached upstream to the side of the open reading frame(ccdB-Myc coding complex) and RBS (AAGGAG). The terminator BBa_B1006 is attached downstream of the sequence. |
===Characterization=== | ===Characterization=== |
Revision as of 05:38, 9 October 2022
pLac+RBS+CcdB-Myc+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.
Construct Design
We attached a Myc epitope Tag downstream of the ccdB sequence for purification purposes. A pLac promoter is attached upstream to the side of the open reading frame(ccdB-Myc coding complex) and RBS (AAGGAG). The terminator BBa_B1006 is attached downstream of the sequence.
Characterization
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 271