pDL52 (clone76)
(Plasmid #188579)

• Purpose: High-yield production of coenzyme F420 in E. coli
• Depositing Lab: Gerald Lackner
• Publication: Last et al Metab Eng. 2022 Jul 18. pii: S1096-7176(22)00093-3. doi: 10.1016/j.ymben.2022.07.006.

Backbone

• Vector backbone: pET28a
• Backbone manufacturer: Novagen
• Backbone size w/o insert (bp): 5021
• Total vector size (bp): 13300
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: For cloning DH5alpha is fine. For expression, E. coli BL21(DE3) is necessary.
• Copy number: High Copy

Gene/Insert 1

• Gene/Insert name: ribA
• Species: E. coli
• Mutation: Codon optimization, synthetic ribosome binding sites
• Promoter: T7 variants

Cloning Information for Gene/Insert 1

• Cloning method: Ligation Independent Cloning
• 5′ sequencing primer: n/a

Gene/Insert 2

• Gene/Insert name: ribD
• Species: E. coli
• Mutation: Codon optimization, synthetic ribosome binding sites

Gene/Insert 3

• Gene/Insert name: yigB
• Species: E. coli
• Mutation: Codon optimization, synthetic ribosome binding sites

Gene/Insert 4

• Gene/Insert name: fbiC
• Species: Mycetohabitans rhizoxinica
• Mutation: Codon optimization, synthetic ribosome binding sites

Gene/Insert 5

• Gene/Insert name: cofC
• Species: Methanocaldococus jannaschii
• Mutation: Codon optimization, synthetic ribosome binding sites

Gene/Insert 6

• Gene/Insert name: cofD
• Species: Methanocaldococus jannaschii
• Mutation: Codon optimization, synthetic ribosome binding sites

Gene/Insert 7

• Gene/Insert name: cofI
• Species: Thermomicrobium roseum
• Mutation: Codon optimization, synthetic ribosome binding sites

Gene/Insert 8

• Gene/Insert name: cofE
• Species: Methanocaldococus jannaschii
• Mutation: Codon optimization, synthetic ribosome binding sites

Resource Information

• Supplemental Documents:
  • pDL52_clone76.gb

Terms and Licenses

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

Depositor Comments

Sequencing of pDL52 plasmids using primers binding the (repetitive) T7 promotor region is only possible after the separation of the two operons e.g. by restriction digestion with HindIII and NcoI followed by purification of the two fragments from preparative agarose gels.