DNA

Part:BBa_K4361006

Designed by: Lars van den Biggelaar   Group: iGEM22_TUDelft   (2022-08-21)
Revision as of 07:35, 10 October 2022 by Larsvdb (Talk | contribs)


BlcR-binding oligo, 51 bp, swapped IRs

BlcR is a transcription factor originating from the bacterium Agrobacterium tumefaciens (Part:BBa_K4361100). In a homodimer state it contains a single DNA-binding domain that specifically binds one of two DNA sequences. Both sequences are so-called inverted repeat pairs (IRs), short DNA sequences whose ends are reverse complements of each other. For the Blc operator, these sequences are 'ACTCTAATgATTCAAGT' (IR1) and 'ATTAGttgaactCTAAT' (IR2), as further explained in Part:BBa_K4361001.
This part has been designed to test whether or not the order of IRs influences the binding strength of BlcR to DNA. To create this part, the nucleotides originally designated as belonging to IR2 have been fully replaced by the sequence of IR1, and those of IR1 by IR2. The BlcR-binding domain of this part thus consists of IR2-tca-IR1, where tca is the original 3 nt linker sequence between IRs.

Based off of BBa_K4361001, but swapping both inverted repeat pairs.

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]

Usage and biology

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Results

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Figure 2. Results of the second Tapestation experiment, in which the fraction of DNA bound to BlcR was determined for different types of oligos. The first bar and bottom dashed line represent the results with Part:BBa_K4361000 (scrambled oligo, negative control), the second bar and top dashed line correspond to those with Part:BBa_K4361001 (wildtype oligo, positive control). The third bar depicts the measured fraction of bound DNA for this part.


[edit]
Categories
//awards/part_collection
Parameters
None