pCD-Sub100-Sem1 Sc
(Plasmid #83241)

• Purpose: Expresses the cDHFR-link2-His6-MBP-Sem1 Sc in E.coli
• Depositing Lab: Gali Prag
• Publication: Levin-Kravets et al Nat Methods. 2016 Nov;13(11):945-952. doi: 10.1038/nmeth.4003. Epub 2016 Oct 3.

Backbone

• Vector backbone: pCDF-1
• Backbone size w/o insert (bp): 3709
• Total vector size (bp): 3979
• Modifications to backbone: The pCD-Sub vector was constructed based on a pCDF-duet vector. The T7 promoter was substituted with pLtetO-1. The cDHFR fragment (residues 109-187) was PCR-amplified and sub-cloned under the pLtetO1 promoter at the MluI and AscI endonuclease recognition sites (the AscI site was inserted by PCR). A second linker (linker2; as indicated in Figure 4B); was fused to the N-terminus of MBP (Maltose Binding Protein) and cloned into the AscI site. Substrates were PCR-amplified and sub-cloned into SacII and SpeI sites downstream and in-frame with the cDHFR-linker2-MBP. In some vectors the MBP was removed. Some vectors were prepared by complete chemical synthesis assembly (Gibson et al, 2009).
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Spectinomycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: SEM 1
• Species: S. cerevisiae (budding yeast)
• Entrez Gene: SEM1 (a.k.a. YDR363W-A, DSS1, HOD1)
• Tag / Fusion Protein:
  • cDHFR-link2-His-mbp (N terminal on insert)

Terms and Licenses

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