pSMART-ala10
(Plasmid #87135)

• Purpose: NADPH dependent Alanine Dehydrogenase, Low Phosphate Dependent E. coli Expression
• Depositing Lab: Michael Lynch
• Publication: Menacho-Melgar et al Biotechnol Bioeng. 2020 May 22. doi: 10.1002/bit.27440.

Backbone

• Vector backbone: pSMART-HCKan (AF532107.1)
• Backbone manufacturer: Lucigen (Middleton, WI)
• Backbone size w/o insert (bp): 1800
• Total vector size (bp): 6256
• Vector type: Bacterial Expression

Growth in Bacteria

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

Gene/Insert 1

• Gene/Insert name: NADPH Alanine Dehydrogenase
• Alt name: ald*
• Species: Synthetic
• Insert Size (bp): 1200
• Mutation: D196A, L197R
• Entrez Gene: ald (a.k.a. BSU31930)
• Promoter: waaHp from E. coli as part of operon with ald* and alaE

Cloning Information for Gene/Insert 1

• Cloning method: Gibson Cloning
• 5′ sequencing primer: CAG TCC AGT TAC GCT GGA GTC
• 3′ sequencing primer: GGT CAG GTA TGA TTT AA A TGG TCA GT

Gene/Insert 2

• Gene/Insert name: alanine exporter
• Alt name: alaE
• Species: Synthetic
• Entrez Gene: alaE (a.k.a. b2670, ECK2664, JW2645, ygaW)
• Promoter: waaHp from E. coli as part of operon with ald* and alaE

Cloning Information for Gene/Insert 2

• Cloning method: Gibson Cloning
• 5′ sequencing primer: CAG TCC AGT TAC GCT GGA GTC
• 3′ sequencing primer: GGT CAG GTA TGA TTT AA A TGG TCA GT

Terms and Licenses

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