This part is a Loop Type IIS Golden Gate adapter that contains a BsaI and a SapI restriction sites.
Usage and Biology
The Loop Type IIS cloning sites (triangles in figure 1) are a combination of BsaI and SapI restriction sites each with different cutting sites that generate well defined overhangs (circles in Figure 1 and 2).
A total of 50 combinations are theoretically possible and this part is one of them: the loop F-Beta (Figure 2).
Blue triangle: SapI fixation site (GCTCTTC) on the complementary strand
Red triangle: BsaI fixation site (GGTCTC)
Blue circle: overhang formed by SapI after cutting (GCA)
Yellow circle: overhang formed by BsaI after cutting (CGCT)
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).
 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
- 10COMPATIBLE WITH RFC
- 12Illegal NheI site found at 9
- 21COMPATIBLE WITH RFC
- 23COMPATIBLE WITH RFC
- 25COMPATIBLE WITH RFC
- 1000Illegal BsaI site found at 1
Illegal SapI.rc site found at 17