DNA

Part:BBa_K4361000

Designed by: Lars van den Biggelaar   Group: iGEM22_TUDelft   (2022-08-21)
Revision as of 14:23, 6 October 2022 by Larsvdb (Talk | contribs)


BlcR-binding oligo, 51 bp, scrambled

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. This is further described in the wildtype oligo, [[Part:BBa_K4361001]]. To test the binding strength of BlcR to its operator sequence and variations thereof, one needs a negative control. To create the negative control, the 51 nt sequence of the wildtype oligo was scrambled using the Genscript Sequence Scramble tool, set to species 'Human'. This tool randomizes the order of the nucleotides of the input sequence, resulting in this part. As a scrambled variant of the original sequence, it has the same amount of each nucleotide, but lacks the specific binding sequence which is otherwise recognized by BlcR. As such, BlcR should not be able to specifically bind this molecule, allowing for its use as a negative control during measurements.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal SpeI site found at 37
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal SpeI site found at 37
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal SpeI site found at 37
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal SpeI site found at 37
  • 1000
    COMPATIBLE WITH RFC[1000]

Usage and Biology

===Results===
In vivo characterization of GBL / GHB sensor with strain containing BBa_K1758377. All experiments were perfomed as triplicates. All samples except "control, not induced" were induced to express T7 polymerase at OD600 = 0.7-0.8 . Control refers to a strain carrying this part in pSB1C3.

[edit]
Categories
//awards/part_collection
Parameters
None