DNA

Part:BBa_K4636000:Design

Designed by: Ping-yu Yang   Group: iGEM23_NTHU-Taiwan   (2023-09-12)


Modified T7 promoter


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]


Design Notes

Refer to previous study, we add four parts of sequence to enhance the reverse transcription ability. Below is the four parts :

(1) Upstream of T7 promoter from -18 to -21 : With highly AT rich. It can increase binding affinity of the T7 polymerase for the DNA template.

(2) Upstream of T7 promoter from -22 to -27 : With highly GC rich. It can increase binding affinity of the T7 polymerase for the DNA template.

(3) Downstream of T7 promoter from +1 to +3 : With triple G (GGG) sequence, It can increase binding affinity of the T7 polymerase for the DNA template. (Reverse transcription start from the first G)

(4) Downstream of T7 promoter from +4 to +8 : With highly AT rich. It can help DNA template to form a initiation bubble which increases the initially transcription ability.



Source

Maximizing transcription of nucleic acids with efficient T7 promoters : https://www.nature.com/articles/s42003-020-01167-x

References