pMT1002
(Plasmid #8621)

• Depositing Lab: Robert Hartley
• Publication: Hartley et al Protein Peptide Letters 1996 3:225-231

Backbone

• Vector backbone: partial pUC19
• Backbone size w/o insert (bp): 1829
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: barnase, barstar
• Species: Bacillus amyloliquefaciens
• Insert Size (bp): 1860
• Mutation: CI157 lambda repressor gene has two HindIII sitesd removed by silent mutations.
• GenBank ID: M14442
• Tag / Fusion Protein:
  • as pMT416 (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoR1 (not destroyed)
• 3′ cloning site: HindIII (not destroyed)
• 5′ sequencing primer: na

Terms and Licenses

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

Depositor Comments

Barnase production plasmid. The vector is the same as that of pMT416. Provides higher yields of barnase than pMT413 or pMT416. This is pTN441 [Okorokov, A.L., Hartley, R.W. and Panov, A,L, Protein Express. and Purif.
5, 547-552 (1994)
] with two HindIII sites in the CI157 gene removed by silent mutations. 900-86: The temperature sensitive repressor gene(CI157) from lambda phage, followed(3')
by its own promoter and, in the forward direction, the PR promoter for barnase. Two silent mutations in this
gene have removed HindIII sites.
901-1872:barnase & barstar genes as in pMT413 & pMT416 from BamH1 site to HindIII site. The copy number is also controlled by the lambda repressor, so that both copy number and barnase production are minimal at 30^o and high at 37^o.