pCR182
(Plasmid #240642)

• Purpose: Template for in vitro transcription of AmNeonGreen mRNA using SP6 polymerase.
• Depositing Lab: John Pringle
• Publication: Aiptasia Genetic Tools Plasmids (unpublished)

Backbone

• Vector backbone: pCS107
• Backbone manufacturer: Baker et al., 1999
• Backbone size w/o insert (bp): 4108
• Total vector size (bp): 4131
• Vector type: Vertebrate gene expression and general in vitro transcription.

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: AmNeonGreen
• Species: Synthetic
• Insert Size (bp): 711
• Mutation: Codon-optimized for Exaiptasia diaphana, amino acid sequence identical to original mNeonGreen.
• GenBank ID: KC295282.1
• Promoter: SP6

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: Pcmv_end_seq_fwd (GCAAATGGGCGGTAGGCG)
• 3′ sequencing primer: M13 Reverse (-27) (CAGGAAACAGCTATGAC)

Resource Information

• Supplemental Documents:
  • pCR182.gb
• A portion of this plasmid was derived from a plasmid made by: pCS107 backbone: Baker JC, Beddington RSP, Harland RM. 1999. Wnt signaling in Xenopus embryos inhibits Bmp4 expression and activates neural development. Genes Dev. 13(23):3149–3159. doi:10.1101/gad.13.23.3149. mNeonGreen: Shaner NC, Lambert GG, Chammas A, Ni Y, Cranfill PJ, Baird MA, Sell BR, Allen JR, Day RN, Israelsson M, et al. 2013. A bright monomeric green fluorescent protein derived from Branchiostoma lanceolatum. Nat Methods. 10(5):407–409. doi:10.1038/nmeth.2413.

Terms and Licenses

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