Part:BBa_K5136044

pColE408-RBS

Biology

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.

Usage

pColE408-RBS was used to initiate the expression of ccdB, CcdB locks DNA gyrase together with broken double-stranded DNA, ultimately leading to cell death. At the same time, we used pColE408-RBS to initiate the expression of aca2 and inhibit the expression of the underlying CcdA protein. We used LexRO, pHybrid 2)-114 version, SD7, ccdA and ccdB to construct the regulation system and obtained the composite part BBa K5136231 , which was assembled on the expression vector pSB4A5.

Characterization

Agarose gel electrophoresis (AGE)
When we were building this circuit, colony PCR was used to certify the plasmid was correct. We got the target fragment-****bp 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
  COMPATIBLE WITH RFC[1000]