PIA586
(Plasmid #104399)

• Purpose: vector for expression of single E.coli RNAP subunit
• Depositing Lab: Irina Artsimovitch
• Publication: Svetlov et al Methods Mol Biol. 2015;1276:13-29. doi: 10.1007/978-1-4939-2392-2_2.

Backbone

• Vector backbone: pET28b
• Backbone size w/o insert (bp): 5368
• Total vector size (bp): 7153
• Modifications to backbone: filled in XhoI site in pET28B, recloned XbaI-HindIII fragment from pET70-σ70 into pIA585 = unique XhoI site in rpoD
• Vector type: Bacterial Expression

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: rpoD
• Species: E. coli
• Promoter: T7
• Tag / Fusion Protein:
  • His6 (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Xba 1 (unknown if destroyed)
• 3′ cloning site: Hind III (unknown if destroyed)
• 5′ sequencing primer: T7 TAATACGACTCACTATAGGG

Terms and Licenses

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

Depositor Comments

T7P–His6-rpoD; N-terminally tagged σ70 subunit under control of the T7 gene 10 promoter