Difference between revisions of "Part:BBa K2983013"

Revision as of 18:03, 19 October 2019

Loop F-Gamma

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 F-Gamma (Figure 2).

Figure 2. Loop F-Gamma structure

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

Red triangle: BsaI fixation site (GGTCTC)

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

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).

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

@@ Line 27: / Line 27: @@
-Blue triangle: SapI fixation site (GCTCTTC)
+Blue triangle: SapI fixation site (GCTCTTC on complementary strand)
-Red triangle: BsaI fixation site (GGTCTC on complementary strand)
+Red triangle: BsaI fixation site (GGTCTC)
 Blue circle: overhang formed by SapI after cutting (TAC)
@@ Line 39: / Line 39: @@
-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)
+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.