pReporter_12
(Plasmid #60572)

• Purpose: A reporter plasmid containing the attB and attP sites flanking a green fluorescent protein reporter gene (gfp).
• Depositing Lab: Christopher Voigt
• Publication: Yang et al Nat Methods. 2014 Dec;11(12):1261-6. doi: 10.1038/nmeth.3147. Epub 2014 Oct 26.

Backbone

• Vector backbone: unknown
• Total vector size (bp): 4314
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Chloramphenicol, 25 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH10B
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: Reporter_12
• Alt name: attB-GFPmut3-RiboJ-attP
• Species: Synthetic
• Promoter: BBa_J23119

Cloning Information

• Cloning method: Unknown
• 5′ sequencing primer: GFP-F
• 3′ sequencing primer: GFP-R

Resource Information

• Supplemental Documents:
  • pReporter_12

Terms and Licenses

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

Depositor Comments

This reporter plasmid contains the attB and attP sites flanking a green fluorescent protein reporter gene (gfp). A strong constitutive promoter (BBa_J23119) is placed upstream of the attB site, which transcribes in the opposite orientation as gfp. After the integrase is expressed, the orientation of gfp is inverted and it is transcribed. After inversion, recombined attB and attP sites result in the formation of attL and attR. The attL site is located on the 5'UTR and could impact gfp expression. To insulate against this effect, we included a spacer and the ribozyme RiboJ.
Also contains mRFP in order to aid the gating of cells.