pMJB1
(Plasmid #160429)

• Purpose: (Empty Backbone) Yeast vector for expressing mCherry tagged genes under the GAL1 promoter
• Depositing Lab: Gary Stormo
• Publication: Roy et al Commun Biol. 2020 Dec 11;3(1):761. doi: 10.1038/s42003-020-01494-z.

Backbone

• Vector backbone: pMW102
• Backbone size (bp): 7415
• Modifications to backbone: The His5 terminator site with 5’-NheI and 3’-XhoI restriction sites was synthesized by G-blocks from IDT. The mCherry sequence was cloned out from pMVS124-pACT1 (a generous gift from Max Staller) with a 3’-NheI and 5’-AvrII restriction sites by PCR using. pMW102-empty-MORF was linearized using primers to incorporate PacI and AvrII restriction sites downstream of the GAL1 promoter. One microgram of purified mCherry, His5 terminator and linearized vector backbone were digested with NheI, AvrII, XhoI and PacI for 15 min and gel purified
• Vector type: Yeast Expression
• Promoter: GAL1
• Selectable markers: URA3
• Tag / Fusion Protein:
  • mCherry (C terminal on backbone)

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Unknown

Cloning Information

• Cloning method: Gibson Cloning

Resource Information

• Supplemental Documents:
  • MJB1-mod5.gen.gb
• A portion of this plasmid was derived from a plasmid made by: This vector was modified from pMW102, which is the creation of one of the submitters (M. White). Other elements as described were from plasmid pMVS124-pACT1, a gift from Max Staller.

Terms and Licenses

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

Depositor Comments

Please visit https://www.biorxiv.org/content/10.1101/2020.05.11.089300v1 for bioRxiv preprint.

pMW102: "A systematic screen for transcriptional regulators of the yeast cell cycle", Michael A White , Linda Riles, Barak A Cohen, Genetics 2009 Feb;181(2):435-46. doi: 10.1534/genetics.108.098145.

pMVS124-pACT1: "A High-Throughput Mutational Scan of an Intrinsically Disordered Acidic Transcriptional Activation Domain", MV Staller, et al., Cell Syst. 2018 Apr 25;6(4):444-455.e6. doi: 10.1016/j.cels.2018.01.015.