pAGM22082_cRed
(Plasmid #117225)

• Purpose: pET-28b backbone plasmid, Level 2 Golden Gate expression plasmid.
• Depositing Lab: Sylvestre Marillonnet
• Publication: Pullmann et al Sci Rep. 2019 Jul 29;9(1):10932. doi: 10.1038/s41598-019-47376-1.

Backbone

• Vector backbone: pET28-b
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 5325
• Total vector size (bp): 13411
• Modifications to backbone: Introduced mutations for Type IIs cleavage site removal
• Vector type: Bacterial Expression

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: Canthaxanthin Operon
• Species: Erwinia uredovora, Escherichia Coli, Agrobacterium aurantiacum
• Insert Size (bp): 8086
• Mutation: removed Type IIS restriction sites within the backbone (BsaI and BbsI)
• GenBank ID: D90087.2
• Promoter: T7 RNA Polymerase
• Tag / Fusion Protein:
  • 34 amino acid N-terminal linker, Hexahistidine Tag, T7 Leader Tag (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BstXI (not destroyed)
• 3′ cloning site: BstXI (not destroyed)
• 5′ sequencing primer: GAAGGAGATATACCATGGGCAGCAG
• 3′ sequencing primer: CGTTTAGAGGCCCCAAGGGGTTATG

Resource Information

• A portion of this plasmid was derived from a plasmid made by: canthaxanthin operon as previously described in pAGM4673 (Plasmid #48014)

Terms and Licenses

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