PGR-Blue
(Plasmid #68374)

• Purpose: (Empty Backbone) Terminator quantification vector utilizing Golden Gate Assembly and amilCP. Positive colonies are green and false positives are blue. Terminators are placed between GFP and RFP - induced by arabinose.
• Depositing Lab: Nathan Reyna
• Publication: Reyna Lab - Terminator Verification Plasmid (unpublished)

Backbone

• Vector backbone: pSB1A10
• Backbone manufacturer: http://partsregistry.org/partsdb/get_part.cgi?part=pSB1A10
• Backbone size (bp): 5191
• Modifications to backbone: Added BsaI cut site and sticky ends for directional golden gate assembly. Left facing AmilCP (Blue Chromo Protein) added between GFP/RFP coding sequences.
• Vector type: Bacterial Expression, Synthetic Biology ; Golden Gate Assembly
• Promoter: pBAD

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): JM109
• Copy number: High Copy

Cloning Information

• Cloning method: Restriction Enzyme

Resource Information

• Supplemental Documents:
  • PGR-Blue_Protocol
• A portion of this plasmid was derived from a plasmid made by: Modification of PGR (Voigt Lab): http://www.addgene.org/46002/
• Articles Citing this Plasmid:
  • 3 References

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Instructions on how to use this plasmid can be found in the associated JoVE article: http://www.jove.com/video/54064.

Terminator quantification vector utilizing Golden Gate Assembly and amilCP for easy colony selection. Colonies containing ligated inserts are Green and colonies containing the uncut plasmid are blue. Synthesized terminators must contain left sticky end of 5’ CGAC 3’ and right sticky end of 5’ CCGC 3’. Terminators are placed between GFP and RFP and expression is driven by pBAD arabinose inducible promoter. This is a modification of the PGR plasmid developed by the Voigt Lab.