pHW2159
(Plasmid #255961)

• Purpose: N-terminal TAG-IN donor plasmid for fusion tagging with mCherry (selection marker, SEC flanked by Lox2272)
• Depositing Lab: Han Wang
• Depositing Organization: University of Wisconsin, Madison
  Located in United States of America
• Publication: Wang Lab TAG-IN plasmids (unpublished)

Backbone

• Vector backbone: pFH173
• Vector type: Worm Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): NEB® Turbo competent E. coli
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: FRT-SEC(pDD382)-mCherry-N' linker-SD-rFRT3
• Species: C. elegans (nematode), Synthetic

Cloning Information

• Cloning method: Golden Gate

Terms and Licenses

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

Depositor Comments

For unknown reasons, the DNA construct (pHW2159) seems toxic to the E. coli strain DH5α, often resulting in unexpected mutations. When transforming this plasmid into DH5α, we recommend selecting only small colonies and sequencing the plasmid following fresh miniprep from a bacterial culture to confirm absence of any mutations. To overcome this issue, the deposited bacterial stock of this plasmid is NEB® Turbo competent E. coli, which better tolerates this construct and more reliably yield plasmid DNA with the correct sequence.