Part:BBa_K2983024

Loop E-Alpha

This part is a Loop Type IIS Golden Gate adapter that contains a BsaI and a SapI restriction sites.

Usage and Biology

Figure 1. BsaI and SapI restriction sites (adapted from [1])

The Loop Type IIS cloning sites (triangles in figures 1 and 2) are a combination of BsaI and SapI restriction sites each with different cutting sites that generate well defined overhangs (circles in Figures 1 and 2).

A total of 50 combinations are theoretically possible and this part is one of them: the loop E-Alpha (Figure 2).

Figure 2. Loop E-Alpha structure

Blue triangle: SapI fixation site (GCTCTTC) on the complementary strand

Red triangle: BsaI fixation site (GGTCTC)

Blue circle: overhang formed by SapI after cutting (ATG)

Orange circle: overhang formed by BsaI after cutting (GCTT)

This part was used to define the Loop assembly system dedicated to the oleaginous yeast Yarrowia lipolytica (for details visit https://2019.igem.org/Team:Evry_Paris-Saclay/Design).

References

[1] Pollak B, Cerda A, Delmans M, Álamos S, Moyano T, West A, Gutiérrez RA, Patron NJ, Federici F, Haseloff J. Loop assembly: a simple and open system for recursive fabrication of DNA circuits. New Phytol (2019) 222, 628-640.

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
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 1
  Illegal SapI.rc site found at 17