pNAS1B
(Plasmid #61968)

• Purpose: Duet vector for co-expression of GFP and mCherry from PBAD and PLtetO promoters, respectively; parent vector for split mCherry assembly vectors
• Depositing Lab: Lynne Regan
• Publication: Sawyer et al ACS Chem Biol. 2014 Nov 21;9(11):2502-7. doi: 10.1021/cb500658w. Epub 2014 Oct 6.

Backbone

• Vector backbone: pNAS1B
• Backbone size w/o insert (bp): 3937
• Total vector size (bp): 5374
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH10B
• Copy number: Low Copy

Gene/Insert 1

• Gene/Insert name: mCherry
• Insert Size (bp): 720
• Promoter: PLtetO

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: KpnI (not destroyed)
• 3′ cloning site: HindIII (not destroyed)
• 5′ sequencing primer: 5'-GATACTGAGCACATCAGCAGGACG-3'
• 3′ sequencing primer: 5'-GTAAGCCAGTATACACTCCGCTAGC-3'

Gene/Insert 2

• Gene/Insert name: GFP
• Insert Size (bp): 717
• Mutation: Serine 65 to Glycine (GGT), Serine 72 to Alanine (GCG); mut3b from Cormack, B.P. et al. 1996 Gene 173, 33-38.
• Promoter: PBAD

Cloning Information for Gene/Insert 2

• Cloning method: Restriction Enzyme
• 5′ cloning site: AatII (not destroyed)
• 3′ cloning site: SacI (not destroyed)
• 5′ sequencing primer: 5'-CCTACCTGACGCTTTTTATCGCAACTC-3'

Resource Information

• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry