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Part:BBa_K2983016

Designed by: Tom Zaplana   Group: iGEM19_Evry_Paris-Saclay   (2019-10-11)


Loop Epsilon-A

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 Epsilon-A (Figure 2).


Figure 2. Loop Espilon-A structure


Blue triangle: SapI fixation site (GCTCTTC)

Red triangle: BsaI fixation site (GGTCTC on complementary strand)

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

Red circle: overhang formed by BsaI after cutting (GGAG)



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.rc site found at 18
    Illegal SapI site found at 1


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