URL: http://www.addgene.org/32602
Proper Citation: RRID:Addgene_32602
Insert Name: myr-Venus
Bacterial Resistance: Ampicillin
Defining Citation: PMID:16604528
Vector Backbone Description: Backbone Size:4700; Vector Backbone:pCAGGS; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
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
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