Part:BBa_K4190011

PGK Terminator Plasmid

Moclo part containing PGK terminator for Golden Gate Assembly.

Usage and Biology

The S. cerevisiae Modular Cloning (MoClo) Library for plasmid construction was built and published in 2015 [1]. This major breakthrough in yeast research enables rapid design of plasmids for protein expression. The MoClo library provides resources for easy, one-step constructions of S. cerevisiae plasmids through GGA. Interchangeable plasmid parts can be specialized allowing for rapid and easy testing of multiple plasmid constructs.

For the construction of our S. cerevisiae genomic insert, we used a GAL1 promoter and a PGK1 terminator. The GAL1 promoter and PGK1 terminator sequences are included as part of the basic block of the yeast MoClo library [1]. They are constructed on an E. coli plasmid containing a Chloramphenicol resistance marker, ColE1 ori, and BsaI binding site. Protein production is initiated by the GAL1 promoter when the S. cerevisiae is grown up in a galactose based liquid media.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal XhoI site found at 1811
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 1797
  Illegal BsaI.rc site found at 147
  

References

[1] M. E. Lee, W. C. DeLoache, B. Cervantes, and J. E. Dueber, “A Highly Characterized Yeast Toolkit for Modular, Multipart Assembly,” ACS Synth. Biol., vol. 4, no. 9, pp. 975–986, Sep. 2015, doi: 10.1021/sb500366v.