Purpose(Empty Backbone) A backbone for adenovirus creation by the AdEasy method, similar to ShuttleCMV (Addgene Plasmid # 16403), except it uses a mouse cofilin promoter (MCP) instead of the CMV promoter
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||50959||Standard format: Plasmid sent in bacteria as agar stab||1||$75 *|
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Backbone manufacturerBert Vogelstein lab (Addgene Plasmid #16402)
- Backbone size (bp) 8507
Modifications to backboneA 1255 bp region of the mouse cofilin 1 gene in BAC RP-23-457M2 just upstream of the start codon was amplified by PCR using the following primers containing a KpnI site: 5’ TTTCTAGATGGTACCGCTTCGGCCTCCACCTGG, 3’ TCTTCTAGAGGTACCGGGAGACAGAAAGAGCAACTG. KpnI-cut MCP DNA was then ligated into the KpnI site of pShuttle. A 710 bp polyadenylation sequence from the 3’UTR of the mouse cofilin-1 gene was amplified from the BAC template using PCR primers that contained a BglII site using the following primers: 5’ TTCAAGATCTGCCGTCATTTCCCTGGAGG, 3’ TTCAAGATCTGAGCCCAACTGCCCTGCC. This polyadenylation sequence was also cloned into the BglII site of pShuttle to generate pShuttle-MCP.
Vector typeMammalian Expression, Adenoviral
- Promoter uses portion of the mouse cofilin gene as a mouse cofilin promoter (MCP)
Growth in Bacteria
Bacterial Resistance(s)Kanamycin, 50 μg/mL
Copy numberLow Copy
- Cloning method Restriction Enzyme
- 5′ sequencing primer Encap-F (5'-TTTGGGCGTAACCGAGTAAG-3')
- 3′ sequencing primer pShuttle-R (5'-CACAATGCTTCCATCAAACG-3') (Common Sequencing Primers)
Terms and Licenses
- Zeocin® is an InvivoGen trademark.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:ShuttleMCP was a gift from James Bamburg (Addgene plasmid # 50959 ; http://n2t.net/addgene:50959 ; RRID:Addgene_50959)
For your References section:A genetically encoded reporter for real-time imaging of cofilin-actin rods in living neurons. Mi J, Shaw AE, Pak CW, Walsh KP, Minamide LS, Bernstein BW, Kuhn TB, Bamburg JR. PLoS One. 2013 Dec 31;8(12):e83609. doi: 10.1371/journal.pone.0083609. PubMed 24391794