pCAG:myr-Venus
(Plasmid #32602)

• Depositing Lab: Anna-Katerina Hadjantonakis
• Publication: Rhee et al Genesis. 2006 Apr;44(4):202-18.

Backbone

• Vector backbone: pCAGGS
• Backbone size w/o insert (bp): 4700
• 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: myr-Venus
• Promoter: CAG

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoR1 (not destroyed)
• 3′ cloning site: EcoR1 (not destroyed)
• 5′ sequencing primer: CTT GAA TTC GCC ACC ATG GGA AGC AGC AAG AGC AAG CCA AAG GTG AGC AAG GGC GAG GAG CTG
• 3′ sequencing primer: GTC ATG AAT TCT TAC TTG TAC AGC TCG TCC

Resource Information

• Articles Citing this Plasmid:
  • 3 References

Terms and Licenses

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

Depositor Comments

To generate myristoylated fluorescent fusion proteins, an N-terminal myristoylation tag
derived from Src was generated by adding the sequences MGSSKSKPK to the N-terminus of
any given fluorescent protein. This was achieved by using the following oligonucleotide: 5′
GFP-MYR-Eco (5′-CTT GAA TTC GCC ACC ATG GGA AGC AGC AAG AGC AAG CCA
AAG GTG AGC AAG GGC GAG GAG CTG). The GFP and Venus coding sequences were
amplified from pEGFP-N1 (BD Biosciences, San Jose, CA) and pCS2-Venus (Nagai et al.,
2002) to generate myr-GFP and myr-Venus, respectively, by high-fidelity polymerase chain
reaction (PCR) using Pfx Polymerase (Invitrogen, La Jolla, CA) with the 5′ myristoylation
primer combined with a 3′-GFP primer (5′-GTC ATG AAT TCT TAC TTG TAC AGC TCG
TCC) primer, respectively. The resulting product was cloned into the EcoRI site of pCAGGS
to generate pCX::myr-EGFP and pCX::myr-Venus