Composite

Part:BBa_K5136235

Designed by: Xiaoxiao Zhang   Group: iGEM24_XMU-China   (2024-09-30)
Revision as of 09:06, 2 October 2024 by Lithiumga (Talk | contribs)


J23106-SD17-LexRO-B0015-pColE408-ccdB-B0014-pHybrid 2)-114 version-ccdA

Biology

LexRO

LexRO is a synthetic light-switchable repressor, based on a novel LOV light sensor domain, RsLOV. In the darkness, LexRO dimerizes and binds to its cognate operator sequence to repress promoter activity. Upon light exposure, the LexRO dimer dissociates, causing dissociation from the operator sequence, and initiates gene expression.

SD7

A SD sequence upstream of LexRO encoding gene. SD7 is a ribosome binding site on the genome , which is capable of recruiting ribosomes in engineered E. coli.

pHybrid 2)-114

pHybrid 2)-114 version is an engineering promoter that is suppressed by the Aca2 repressor, which uses the -35 and -10 regions of J23114.

pColE408-RBS

pColE408-RBS is a combination of the promoter pColE408 and RBS. PcolE408 is a promoter to which LexRO (BBa_K5136042) binds and represses the expression of genes driven by the promoter. The promoter mainly consists of two parts: ColE promoter, which is a strong promoter, and LexA(408)-operator, which is the binding site of LexRO.

ccdB

ccdB gene is located on the Escherichia coli F-factor plasmid and is part of the toxin-antitoxin system encoded by the ccd operon, responsible for plasmid maintenance during cell division. ccdB encodes a toxic protein that acts as a DNA gyrase poison. It can bind DNA gyrase to the broken double-stranded DNA, leading to cell death.

ccdA

ccdA is the gene found within the ccd operon, encoding the antidote protein (CcdA) that protects cells from the toxic effects of CcdB. CcdA protein is easily degraded by Lonprotease.The cell loses the ccdA gene due to the loss of the F plasmid, causing the cell to succumb to the toxicity of CcdB.

Characterization

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal NheI site found at 1610
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 456
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1379


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Categories
Parameters
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