Plasmid_Backbone

Part:BBa_K1680014:Design

Designed by: Katharina Sporbeck   Group: iGEM15_Tuebingen   (2015-09-17)

pRS313 yeast shuttle vector with Biobrick MCS


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2120
    Illegal XbaI site found at 2105
    Illegal SpeI site found at 2097
    Illegal PstI site found at 2083
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2120
    Illegal NheI site found at 1007
    Illegal SpeI site found at 2097
    Illegal PstI site found at 2083
    Illegal NotI site found at 2088
    Illegal NotI site found at 2112
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2120
    Illegal BglII site found at 898
    Illegal BglII site found at 958
  • 23
    INCOMPATIBLE WITH RFC[23]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2120
    Illegal XbaI site found at 2105
    Illegal SpeI site found at 2097
    Illegal PstI site found at 2083
  • 25
    INCOMPATIBLE WITH RFC[25]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2120
    Illegal XbaI site found at 2105
    Illegal SpeI site found at 2097
    Illegal PstI site found at 2083
    Illegal NgoMIV site found at 1748
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal BsaI.rc site found at 3494
    Illegal SapI site found at 2411
    Illegal SapI site found at 4515


Design Notes

This part is derived from the pRS313 vector (Sikorski 1989). It is mutagenised to be compatible with RFC10 and the wild type MCS was switched for a RFC10 MCS. The biobrick MCS is flanked by ApaI (Prefix) and SacI (Suffix) restriction sites. These can be used together with either EcoRI or PstI site to introduce a promoter or terminator stably into the vector without disturbing the RFC10 standard.


Sequence of the new RFC10 MCS:

GGGCCCACGTGAATTCGCGGCCGCTTCTAGAGTACTAGTAGCGGCCGCTGCAGACGTGAGCTC

Source

Derived from pRS313 Part:BBa_K950008 (Sikorski 1989).

References

Sikorski, Robert S., and Philip Hieter. "A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae." Genetics 122.1 (1989): 19-27.