pBS MCK CRE
(Plasmid #12529)

• Depositing Lab: Ronald Kahn
• Publication: Bruning et al Mol Cell. 1998 Nov . 2(5):559-69.

Backbone

• Vector backbone: pBluescript KS
• Backbone size w/o insert (bp): 9500
• Vector type: Cre/Lox ; transgenic construct

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: Cre recombinase
• Insert Size (bp): 1300
• Mutation: Cre is driven by the Muscle Creatine Kinase promoter

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: SacI (destroyed during cloning)
• 3′ cloning site: SacII (destroyed during cloning)
• 5′ sequencing primer: M13 reverse

Resource Information

• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

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.