|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||26818||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
- Backbone size w/o insert (bp) 5400
Vector typeMammalian Expression
Growth in Bacteria
Copy numberHigh Copy
SpeciesH. sapiens (human)
Insert Size (bp)1504
MutationSequence begins from second ATG; Synthetic UTR sequences added to c-MYC ORF
Entrez GeneMYC (a.k.a. MRTL, MYCC, bHLHe39, c-Myc)
- Cloning method Restriction Enzyme
- 5′ sequencing primer CMV Forward
- 3′ sequencing primer TK polyA reverse (Common Sequencing Primers)
Note that orientation of the insert in pcDNA does not matter -- they were cloned by TA-cloning, and either orientation is fine.
Comment from lab: "with reference to NM_002467.4 sequence we have two mutations at following positions: 172 (A to G) and 174 (C to G) there is some confusion as to what the ref seq is exactly but these are the constructs that we've been using successfully."
Please note that the 5'UTR and 3'UTR sequences are synthetic and were added to the c-MYC ORF as described in Figure S1 and Table S2 of Warren et al., Cell Stem Cell. 2010. These UTR sequences are not endogenous c-MYC UTR sequences.
The 5' UTR sequence (including T7 promoter) is: TGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAAATAAGAG
The 3' UTR sequence is: GCTGCCTTCTGCGGGGCTTGCCTTCTGGCCATGCCCTTCTTCTCT
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:pcDNA3.3_c-MYC was a gift from Derrick Rossi (Addgene plasmid # 26818 ; http://n2t.net/addgene:26818 ; RRID:Addgene_26818)
For your References section:Highly Efficient Reprogramming to Pluripotency and Directed Differentiation of Human Cells with Synthetic Modified mRNA. Warren L, Manos PD, Ahfeldt T, Loh YH, Li H, Lau F, Ebina W, Mandal PK, Smith ZD, Meissner A, Daley GQ, Brack AS, Collins JJ, Cowan C, Schlaeger TM, Rossi DJ. Cell Stem Cell. 2010 Sep 29. ():. 10.1016/j.stem.2010.08.012 PubMed 20888316