pBS MCK CRE
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||12529||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
Vector backbonepBluescript KS
- Backbone size w/o insert (bp) 9500
Vector typeCre/Lox ; transgenic construct
Growth in Bacteria
Copy numberHigh Copy
Gene/Insert nameCre recombinase
Insert Size (bp)1300
MutationCre is driven by the Muscle Creatine Kinase promoter
- Cloning method Restriction Enzyme
- 5′ cloning site SacI (destroyed during cloning)
- 3′ cloning site SacII (destroyed during cloning)
- 5′ sequencing primer M13 reverse (Common Sequencing Primers)
Terms and Licenses
- Not Available to Industry
To create a construct for muscle-specific expression of the Cre recombinase, a 6.5 kb XhoI fragment containing 3 kb of the MCK promoter and enhancer 1, untranslated exon 1, 3 kb of intron 1 including the enhancer region E2, and the first 16 bp of exon 2 was subcloned into the XhoI site of pBluescriptKS to result in pBSMCK (see Addgene plasmid #12528 for details. Plasmid 12528 contains a polyA sequence, whereas the base vector that MCK cre was built from does not - the polyA was added using a different cloning strategy as described below.). A 1.3 kb blunt-ended SacI/SacII fragment containing a modified Cre cDNA with a SV-40 large T antigen nuclear localization signal inserted in-frame after the ATG of Cre and a poly(A) signal was inserted into the blunt-ended PstI site of pBSMCK to result in pMCKNlsCrepolyA.
Please see "Author's Map" for a summary of the Kahn Lab's sequencing results.
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:pBS MCK CRE was a gift from Ronald Kahn (Addgene plasmid # 12529 ; http://n2t.net/addgene:12529 ; RRID:Addgene_12529)
For your References section:A muscle-specific insulin receptor knockout exhibits features of the metabolic syndrome of NIDDM without altering glucose tolerance. Bruning JC, Michael MD, Winnay JN, Hayashi T, Horsch D, Accili D, Goodyear LJ, Kahn CR. Mol Cell. 1998 Nov . 2(5):559-69. 10.1016/S1097-2765(00)80155-0 PubMed 9844629