pUS252
(Plasmid #127674)

• Purpose: Expresses fuGFP constitutively in E.coli cells. Designed to act as modular template for cutting out or amplifying fuGFP to place in other plasmid backbones
• Depositing Lab: Nicholas Coleman
• Publication: A new open-source superfolding green fluorescent protein: Free-Use GFP (fuGFP) (unpublished)

Backbone

• Vector backbone: pK18
• Backbone size w/o insert (bp): 2620
• Total vector size (bp): 3359
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: Based on what we know from other high-copy GFP-expressing vectors, there is a chance of vector instability (deletions / loss of GFP gene), although we havent seen this for this plasmid. Reducing growth temperature to 30 C can help to prevent this, since it lowers the copy number of the plasmid.
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Free Use GFP (fuGFP)
• Species: Synthetic
• Insert Size (bp): 739
• Promoter: lac

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI, EcoRI, XbaI, XhoI, NdeI (not destroyed)
• 3′ cloning site: HindIII, PstI, SphI, NheI (not destroyed)
• 5′ sequencing primer: M13 rev
• 3′ sequencing primer: M13 fwd

Resource Information

• Supplemental Documents:
  • pUS252 fuGFP notes

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Industry MTA

Depositor Comments

The creators (Nick Coleman & Mark Somerville) request that their work is cited once published. Please follow Addgene's "How to Cite" guidelines below.