pMOD4-mT2A-mutated_rpsL-BSD-F
(Plasmid #182338)

• Purpose: Template plasmid for PCR amplification of initial recombineering cassette
• Depositing Lab: Mario Capecchi
• Publication: Additional gene targeting vector construction plasmids (unpublished)

Backbone

• Vector backbone: pMOD4-galK-G
• Backbone manufacturer: Alfred L. Fisher
• Vector type: Bacterial Expression, Mouse Targeting ; FLP / FRT
• Selectable markers: Blasticidin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): EC100D pir-116 E. coli (Epicentre)
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: BSD
• Species: Synthetic

Resource Information

• Supplemental Documents:
  • * 2 pMOD4-mT2A-rpsL-BSD-F.gb
• A portion of this plasmid was derived from a plasmid made by: BSD is from Stewart A. Anderson.

Terms and Licenses

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

Depositor Comments

PCR amplify around the 5' T2A and 3' FRT site, making sure to add a PacI site at the 3' of the FRT site.

As suggested in the plasmid name, due to a PCR error, the rpsL open reading frame has a 1-bp deletion that results in a frame shift and as a consequence premature stop codons. Despite this, the downstream BSD open reading frame is translated from the bicistronic RNA in E. coli without problems. These mean that the mutated rpsL cannot be used for streptomycin counterselection, but the unmutated BSD can and was successfully used several times to confer blasticidin resistance for selection after recombineering.