Plasmid_Backbone

Part:BBa_K2077000

Designed by: Danielle Bauhan, Alexander Lacrampe, Joseph Lee, and Richard Anthony   Group: iGEM16_RHIT   (2016-08-09)
Revision as of 13:15, 13 August 2016 by RicRHIT (Talk | contribs)


pSB416-GPD: URA3-selectable S. cerevisiae Expression Vector (RFC 10)

This is a RFC10 standard yeast expression vector adapted from the widely used plasmid p416-GPD. The MCS of p416-GPD was replaced with the standard BioBrick prefix and suffix and an illegal PstI restriction site near the URA3 promoter of p416-GPD was removed. In the resultant plasmid, pSB416-GPD (5743 bp), the BioBrick prefix and suffix is situated between the strong constitutive yeast GPD promoter (TDH3) and CYC1 terminator allowing for facile cloning and expression of translational units in yeast. Standard VF2 and VR primer binding sites are included and M13 primer binding sites bracket the promoter and terminator.

The plasmid has a ColE1 origin and ampicillin resistance for replication in E. coli. It also contains a yeast CEN/ARS and URA3-selectable marker to provide high fidelity maintenance and low copy number in S. cerevisiae.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 5722
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal NotI site found at 9
    Illegal NotI site found at 5728
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 5722
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found at 5722
    Illegal suffix found at 2
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found at 5722
    Plasmid lacks a suffix.
    Illegal XbaI site found at 5737
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal NgoMIV site found at 603
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal BsaI site found at 3703
    Illegal BsaI.rc site found at 1195
    Illegal SapI site found at 1347
    Illegal SapI.rc site found at 2681
    Illegal SapI.rc site found at 4785


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Parameters
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