Part:BBa_K5232004

ds-template

Description: It has a sequence that is fully complementary to primer3 ^[1], with the middle portion having a recognition sequence for the Cas3-Cascade/crRNA complex. Usage: <img src="https://static.igem.wiki/teams/5232/wiki/mechanism/mechanism-3-ds-amp.svg" alt="hogehoge"> Cas3 recognizes a double-stranded PAM sequence and a single-stranded spacer sequence for activation. It has been reported by Yoshimi et al. that even partial PAM sequences combined with a complete spacer sequence can still elicit some level of activity ^[2]. Therefore, a sequence complementary to the spacer sequence was incorporated into the ds-template.In other words, the ds-template sequentially contains a complementary sequence to the primer, a PAM sequence, and one strand of the complementary spacer sequence, starting from the 3' end. <img src="https://static.igem.wiki/teams/5232/wiki/result/result-ds-cas3.svg" alt="hogehoge"> When ds Amp and Cas3 system were connected, no increase in NC fluorescence intensity was observed, and the graph showed a slope dependent on target concentration.

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]