pCALNL-BsaI-EGFP
(Plasmid #123136)

• Purpose: Golden Gate acceptor vector for the assembly of EGFP-based mammalian recombinase reporters.
• Depositing Lab: David Liu
• Publication: Bessen et al Nat Commun. 2019 Apr 26;10(1):1937. doi: 10.1038/s41467-019-09987-0.

Backbone

• Vector backbone: pCANLN
• Backbone size w/o insert (bp): 5000
• Total vector size (bp): 6500
• Vector type: Mammalian Expression, Synthetic Biology

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Mach1
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: EGFP, mRFP
• Insert Size (bp): 1000
• Promoter: cBA

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Esp3I (destroyed during cloning)
• 3′ cloning site: Esp3I (destroyed during cloning)
• 5′ sequencing primer: TCTGCTAACCATGTTCATGCCTTCTTCTTT
• 3′ sequencing primer: CCGTCGACTGCAGAATTCAATTTAAATCGT

Resource Information

• Supplemental Documents:
  • pCALNL-BsaI-EGFP.gb

Terms and Licenses

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

Depositor Comments

Golden gate acceptor vector which, in conjunction with pBT100-neo donor vector and two dsDNA recombinase targets, assembles to form a fluorescent EGFP transfectable reporter. Acceptor vector has mRFP dropout cassette for ease of visualizing transformants that have received the donor cassette (white colonies).