SKA974
(Bacterial strain #80403)

• Purpose: JT2-based E. coli host strain for optogenetic control of cell growth.
• Depositing Lab: Mustafa Khammash
• Publication: Milias-Argeitis et al Nat Commun. 2016 Aug 26;7:12546. doi: 10.1038/ncomms12546.

Backbone

• Vector backbone: none

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): SKA974 (JT2 ΔmetE Tn7::PcpcG2Δ59-metE)
• Growth instructions: Grow in LB or minimal media supplemented with methionine.
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: none

Resource Information

• Supplemental Documents:
  • PCR primers for verification of SKA974
• A portion of this plasmid was derived from a plasmid made by: SKA974 was derived from strain JT2 (Tabor JJ, Levskaya A, and Voigt CA. 2011. J Mol Biol, 405(2): 315-324.)

Terms and Licenses

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

Depositor Comments

Genotype: ΔmetE Tn7::PcpcG2Δ59-metE

Precursor strain: JT2 (Tabor JJ, Levskaya A, and Voigt CA. 2011. J Mol Biol, 405(2): 315-324.) Endogenous metE has been deleted and metE under the cpcG2Δ59 promoter has been integrated into the Tn7 attachment site on the chromosome.

metE was deleted from strain JT2 using λ-Red recombination (Baba T, et al. 2006. Mol Syst Biol, 2. doi: 10.1038/msb4100050). PcpcG2Δ59-metE was then integrated into the Tn7 attachment site by tri-parental mating with Addgene plasmid pSKA397 (Plasmid #80380), helper plasmid pTNS3 (Choi KH, et al. 2008. Applied and Environmental Microbiology, 74(4): 1064-1075.), and E. coli mating strain MFDpir (Ferrières L, et al. 2010. Journal of Bacteriology, 192(24): 6418-6427.).