pMT413
(Plasmid #8606)

• Depositing Lab: Robert Hartley
• Publication: Hartley et al J Mol Biol 1988 Aug 20;202(4):913-5.

Backbone

• Vector backbone: pUC19
• Backbone size w/o insert (bp): 2641
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: Requires strain with the lacI^Q gene (such as DH5alpha, XL1-blue, SURE or HB107)
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: barnase, barstar
• Species: Bacillus amyloliquefaciens
• Insert Size (bp): 1071
• GenBank ID: M14442 M15545
• Tag / Fusion Protein:
  • phoA signal sequence to barnase (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoRI (not destroyed)
• 3′ cloning site: HindIII (not destroyed)
• 5′ sequencing primer: M13F
• 3′ sequencing primer: M13R

Terms and Licenses

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

Depositor Comments

Barnase production (secretion)plasmid. Xba1 site is joint between barnase 3' and barstar 5' sequences. Requires an E. coli host carrying the lacI^Q gene.
Barnase and barstar genes from B. amyloliquefaciens strain H. 7-117: tac promoter and RBS.
132-194: Structural gene for propeptide phoA signal sequence.
195-628: DNA from B. amyloliquefaciens.
195-527: Structural gene for mature barnase. Genbank #M14442.
528-563: Transcription terminator.
635-1077: DNA from B. amyloliquefaciens.
763-1035: Structural gene for barstar. Genbank #X15545. Grown in Super Broth at 37oC, yield on the order of 20 mg/l is maintained from about 1 to 4 hours after growth stops.