pMT440
(Plasmid #8609)

• Depositing Lab: Robert Hartley
• Publication: Yazynin et al Gene 1996 Feb 22;169(1):131-2.

Backbone

• Vector backbone: partial pUC19, as pMT416
• Backbone size w/o insert (bp): 1747
• Vector type: Bacterial Expression

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: barnase, barstar
• Species: Bacillus amyloliquefaciens
• Insert Size (bp): 1071
• Mutation: V36 codon of barnase of pMT416 replaced by polylinker of pUC19. Original EcoR1, BamH1, Xba1- Pst1 and HindIII sites first removed by cutting, blunting and religating.
• GenBank ID: M14442 X15545
• Tag / Fusion Protein:
  • as pMT416

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoRI (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

General purpose conditionally lethal vector with a multiple cloning (MCS)
site for positive selection of cloned inserts. As pMT416 but with MCS from pUC19
inserted within barnase gene. Can be carried in E. coli carrying the lacI^Q gene
in the absence of IPTG. With IPTG it becomes
nonlethal only with DNA fragments inserted into the MCS. Except for Sac1,
MCS sites found elsewhere have been removed . 9-100: tac promoter and RBS.
116-178: Structural gene for propeptide phoA signal sequence.
179-562: Structural gene for barnase with nondecapeptide insert.
283-340: MCS insert.
775-1044: Barstar structural gene