Plasmid_Backbone

Part:BBa_K590011

Designed by: Michael Brasino, Alicia Wong, Rashmi Ravichandran   Group: iGEM11_Washington   (2011-09-15)
Revision as of 23:20, 28 September 2011 by Swanson (Talk | contribs) (Usage and Biology)

pGA1C3, Gibson assembly plasmid (bglBrick) with pLac-GFP insert

This Gibson Cloning friendly 1C3 plasmid backbone was made by [http://2011.igem.org/Team:Washington UW iGEM Team 2011] as part of the [http://2011.igem.org/Team:Washington/Magnetosomes/GibsonVectors Gibson Assembly toolkit].

Usage and Biology

This is a high copy plasmid backbone which has Chloramphenicol resistance, with pLac promoter and GFP. This is a high efficiency Gibson Assembly vector.

Characterization

A sister plasmid (pGA1A3) with the same BglBrick prefix and suffix regions has been shown to have much higher efficiency than the equivalent pSB vector. On a average of 6 plates, the Gibson efficiency was determined to be 0.991129032 ~ 99%!. See [http://2011.igem.org/Team:Washington/Magnetosomes/GibsonResults#Comparison_between_pGA_and_pSB_vectors Gibson Assembly efficiency assay] page for details on the protocol and efficiency measurements.

Washington_pGAefficiency_summary.jpg

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2051
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2051
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2051
    Illegal BglII site found at 2066
    Illegal BamHI site found at 1
    Illegal XhoI site found at 16
    Illegal XhoI site found at 1035
    Illegal XhoI site found at 1927
  • 23
    INCOMPATIBLE WITH RFC[23]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2051
  • 25
    INCOMPATIBLE WITH RFC[25]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2051
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.


[edit]
Categories
Parameters
copies100-300
originpMB1
resistanceChloramphenicol