human EB1 fused to monomeric RFP
(Plasmid #39323)

• Depositing Labs: Tim Mitchison, Jennifer Tirnauer
• Publication: Tirnauer lab EB1 plasmids (unpublished)

Backbone

• Vector backbone: pEGFP-N1
• Backbone manufacturer: clontech
• Backbone size w/o insert (bp): 3982
• Total vector size (bp): 5461
• Modifications to backbone: GFP cut out to swap in mRFP as a BamH1 - Not1 fragment. The mRFP is a PCR product from R Tsien's mRFP plasmid.
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Unknown

Gene/Insert 1

• Gene/Insert name: EB1
• Alt name: MAPRE1
• Species: H. sapiens (human)
• Insert Size (bp): 804
• Entrez Gene: MAPRE1 (a.k.a. EB1)
• Promoter: CMV

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: Xho1 (not destroyed)
• 3′ cloning site: BamH1 (not destroyed)
• 5′ sequencing primer: CMV-F
• 3′ sequencing primer: mCherry-R

Gene/Insert 2

• Gene/Insert name: mRFP
• Alt name: monomeric RFP
• Species: Synthetic
• Insert Size (bp): 675

Cloning Information for Gene/Insert 2

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamH1 (not destroyed)
• 3′ cloning site: Not1 (not destroyed)
• 5′ sequencing primer: n/a
• 3′ sequencing primer: SV40pA-R

Resource Information

• A portion of this plasmid was derived from a plasmid made by: mRFP from Roger Tsien
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

This construct makes very nice comets on growing microtubule tips; mRFP is early generation from ~2002, excitation max 584 nm and emission max 607 nm.