pCM184
(Plasmid #46012)

• Purpose: (Empty Backbone)
• Depositing Lab: Mary Lidstrom
• Publication: Marx et al Biotechniques. 2002 Nov;33(5):1062-7.

Backbone

• Vector backbone: pAYC61
• Backbone manufacturer: Lidstrom Lab (PMID 8021187)
• Backbone size (bp): 6000
• Modifications to backbone: To insert loxP bounded antibiotic resistance gene: The 1.3-kb HincII fragment bearing the kanamycin resistance cassette from pUC4K (Vieira, 1982) was inserted into pLox1 (Palmeros, 2000), which had been cut with XbaI and blunted, to create pCM161. To introduce convenient multiple cloning sites, the loxP-bounded kanamycin cassette of pCM161 was amplified with following primer pair, CM-ufkMCS, 5′-TGACGTCTAGATCTGAATTCAGCTGTACAATTGGTACCATGGATGCATATGGCGGCCGCA-3′, and CM-dfkMCS, 5′-GACTAGTGAGCTCACCGGTTAACACGCGTACGTAGGGCCCGCGGTATCGATAAGCTGGATCC-3′. The resulting 1.4-kb PCR product was purified and cloned into pCR2.1 (Invitrogen, Carlsbad, CA, USA) to create pCM183. To preserve useful cloning sites, pAYC61 was cut with EcoRI and SmaI, blunted using T4 DNA polymerase, and selfligated to produce pCM182. Finally, the 1.4-kb AatII-SpeI fragment from pCM183 containing the loxP-flanked kanamycin cassette was ligated between the AatII and XbaI sites of pCM182 to create pCM184.
• Vector type: Cre/Lox
• Promoter: na

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin and Kanamycin, 100 & 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Unknown

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ sequencing primer: na

Resource Information

• Supplemental Documents:
  • pCM184
• Articles Citing this Plasmid:
  • 3 References

Terms and Licenses

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

Depositor Comments

This allelic exchange vector is used with Cre expression vectors (Addgene plasmids 45863 or 45864) to generate unmarked mutants in a broad bacterial host range.

The full plasmid sequence is approximate and there may be small discrepancies present.