Plasmid_Backbone

Part:BBa_K635002:Design

Designed by: Arjun Athreya   Group: iGEM11_Virginia   (2011-09-27)


Artificial Yeast Chromosome (Ura3 & Amp)


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal NheI site found at 175
    Illegal NheI site found at 3149
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal BglII site found at 4850
    Illegal XhoI site found at 22
  • 23
    INCOMPATIBLE WITH RFC[23]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
  • 25
    INCOMPATIBLE WITH RFC[25]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal NgoMIV site found at 7
    Illegal NgoMIV site found at 4858
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.


Design Notes

The plasmid backbone contained a Gal1/10 bidirectional promoter between the EcoR1 and BamHI sites on the pRS316 plasmid. The plasmid had to be modified for biobrick compatibility by using the HindIII and SacI sites to digest and ligate a gene fragment with the corresponding HindIII and SacI sites, but which also contained the biobrick prefix and suffix in the correct orientation.


Source

The part was modified for biobrick compatibility from a working pRS316 plasmid containing a Gal1/10 bidirectional promoter which was removed through restriction digest.

References