pCMX/cyclinB1-GFP (27)
(Plasmid #26061)

• Depositing Lab: Jonathon Pines
• Publication: Hagting et al Curr Biol. 1999 Jul 1. 9(13):680-9.

Backbone

• Vector backbone: pCMX
• Backbone size w/o insert (bp): 4500
• Vector type: Mammalian Expression

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: Cyclin B1
• Species: H. sapiens (human)
• Entrez Gene: CCNB1 (a.k.a. CCNB)
• Tag / Fusion Protein:
  • GFP (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: HindIII (unknown if destroyed)
• 3′ cloning site: BamHI (unknown if destroyed)
• 5′ sequencing primer: CMV-F
• 3′ sequencing primer: GFP-R

Resource Information

• Articles Citing this Plasmid:
  • 4 References

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

The stop codon of cyclin B1 in the pCMX vector was mutated to a Mfe1 site to link cyclin B1 to MmGFP. Cyclin B1 was fused to the second amino acid residue of MmGFP by a 5 amino acid linker (AGAQF). The construct is cloned as a HindIII-BamHI fragment into pCMX.

The Cyclin insert contains a V5A polymorphism which has no functional consequence. More information on MmGFP can be found in the following reference: Zernicka-Goetz, M., Pines, J., M., Ryan, K., Siemering, K.R., Haseloff, J., and Gurdon, J.B. (1996) ‘An indelible lineage marker for Xenopus using a mutated green fluorescent protein.