Plasmid_Backbone

Part:BBa_K590014

Designed by: Michael Brasino, Alicia Wong, Rashmi Ravichandran   Group: iGEM11_Washington   (2011-09-15)
Revision as of 19:54, 28 September 2011 by Robere (Talk | contribs) (Usage and Biology)

pGA3K3, Gibson assembly plasmid (bglBrick) with pLac-GFP insert

This is a Gibson Cloning friendly 3K3 plasmid backbone that 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 medium copy plasmid backbone that confers kanamycin resistance. It was deposited in the registry with an insert coding for LacI-repressible GFP.

Below is the gel image of the plasmid amplified with universal pGA backbone primers pGAsuffix_fwd and pGAprefix_rev. The band is near the expected length of 2750 bp.

1kb Ladder (left), pGA3k3 (right)

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2729
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2729
    Illegal NheI site found at 1384
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2729
    Illegal BglII site found at 2744
    Illegal BamHI site found at 1
    Illegal XhoI site found at 16
    Illegal XhoI site found at 177
    Illegal XhoI site found at 1020
  • 23
    INCOMPATIBLE WITH RFC[23]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2729
  • 25
    INCOMPATIBLE WITH RFC[25]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2729
    Illegal AgeI site found at 1470
    Illegal AgeI site found at 1793
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal BsaI.rc site found at 2498


[edit]
Categories
Parameters
copies10-30
originp15A
resistanceKanamycin