pRK415-mNeonGreen(inducible expression)
(Plasmid #252211)

• Purpose: Expresses mNeonGreen in a very low copy F-like plasmid under the inducible expression of a tac promoter within E. coli DH10Beta (for cloning)
• Depositing Lab: Cecile Fradin
• Publication: Experimental and simulated FRAP for the quantitative determination of protein diffusion in helical cells (unpublished)

Backbone

• Vector backbone: pRK415
• Backbone manufacturer: Nova Lifetech Inc.
• Backbone size w/o insert (bp): 10416
• Total vector size (bp): 12529
• Modifications to backbone: Additions: mNeonGreen fluorescent protein, tac promoter upstream of mNG, optimized ribosome binding site for Paramagnetospirillum, Lac repressor and promoter system
• Vector type: Bacterial Expression, Synthetic Biology
• Selectable markers: Tetracycline

Growth in Bacteria

• Bacterial Resistance(s): Tetracycline, 10 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): E. coli DH10 Beta
• Growth instructions: LB Miller, 250-300 RPM of shaking
• Copy number: Low Copy

Gene/Insert 1

• Gene/Insert name: mNeonGreen
• Species: Synthetic; Branchiostoma lanceolatum
• Insert Size (bp): 711
• GenBank ID: KC295282.1
• Promoter: tac

Cloning Information for Gene/Insert 1

• Cloning method: Other
• 5′ sequencing primer: CATAATTCGTGTCGCTC
• 3′ sequencing primer: GCCTCTTCGCTATTACG

Gene/Insert 2

• Gene/Insert name: LacI
• Alt name: Lac repressor
• Species: E. coli
• Insert Size (bp): 1083
• Mutation: deleted amino acids 1-3.
• GenBank ID: WP_000248563.1 CP194969.1
• Promoter: LacI promoter

Cloning Information for Gene/Insert 2

• Cloning method: Other
• 5′ sequencing primer: GAGCGACACGAATTATG
• 3′ sequencing primer: caagcacgtccccatg

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • mNeonGreen Label License
• Industry Terms:
  • Not Available to Industry

Depositor Comments

This plasmid was constructed via isothermal DNA assembly that was optimized with a proprietary technique protected by US Provisional (US 63/881,962)