Generator
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Part:BBa_K314203

Designed by: Matthew Coyne, Ingrid Swanson, Jesa Landis, Matthew Harger   Group: iGEM10_Washington   (2010-10-19)
Revision as of 00:50, 21 October 2010 by Hargem (Talk | contribs) (→‎Sequence and Features)

HSL inducible toxin antitoxin, Tse2 + Tsi2

Tse2/Tsi2 w/HSL inducible promoter

This part contains the Tse2/Tsi2 locus from Pseudomonas aeruginosa downstream from an HSL induced promoter( F2620)

Usage and Biology

Tse2 is a toxin that can be exported into gram (-) bacteria by the Type VI secretion system, killing all gram (-) cells not expressing Tsi2 ( the antitoxin of Tse2). Since Tse2 is being coexpressed with Tsi2, cells transformed with this part will not die. Therefore, this part is useful for regulating secretion of Tse2.

Functional Parameters

This part has been tested in pSB3k3. E. coli transformed with this part showed Tse2 production in a broth culture containing HSL at a concentration of 10µM showed Tse2 expression, while no appreciable level of Tse2 was produced in an environment not containing HSL. This was verified by an anti-Tse2 western blot of cellular supernatant of colonies grown up in liquid cultures containing either 10µM HSL or no HSL for a period of approximatively 24 hours.
Cells showed Tse2 production in response to HSL

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
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1283
    Illegal NgoMIV site found at 1340
    Illegal NgoMIV site found at 1465
    Illegal NgoMIV site found at 1795
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1004
    Illegal SapI.rc site found at 1790

====Note: This part does not contain the standard biobricks scar between Lux pr and

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Parameters
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