Difference between revisions of "Part:BBa K2983012"
 
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<partinfo>BBa_K2983012 short</partinfo>
 
<partinfo>BBa_K2983012 short</partinfo>
  
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This part is a Loop Type IIS Golden gate adapter that contains a BsaI and a SapI restriction sites.
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This part is a Loop Type IIS Golden Gate adapter that contains a BsaI and a SapI restriction sites.
  
 
===Usage and Biology===
 
===Usage and Biology===
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A total of 50 combinations are theoretically possible and this part is one of them: the loop Alpha-A (Figure 2).
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A total of 50 combinations are theoretically possible and this part is one of them: the loop Beta-A (Figure 2).
  
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[[File:T--Evry Paris-Saclay--Loop B-Alpha.png|300px|thumb|left| Figure2: Loop Beta-A structure]]
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[[File:T--Evry Paris-Saclay--Loop Beta-A.png|300px|thumb|left| Figure 2. Loop Beta-A structure]]
  
  
 
Blue triangle: SapI fixation site (GCTCTTC)
 
Blue triangle: SapI fixation site (GCTCTTC)
  
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Red triangle : BsaI fixation site (GAGACC)
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Red triangle: BsaI fixation site (GGTCTC on complementary strand)
  
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Blue circle : overhang formed by SapI after cutting (GCA)
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Blue circle: overhang formed by SapI after cutting (GCA)
  
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red circle: overhang formed by BasI  after cutting (GGAG)
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Red circle: overhang formed by BsaI after cutting (GGAG)
  
  
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[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.
 
[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.
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Latest revision as of 17:47, 19 October 2019


Loop Beta-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 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 Beta-A (Figure 2).

Figure 2. Loop Beta-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 (GCA)

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