p5T7-LYCipi-ggpps
(Plasmid #122017)

• Purpose: Expresses genes for lycopene synthesis (crtI, crtB & ipi from Pantoea agglomerans, ggpps from T. canadiensis). IPTG inducible. Spec. resist.
• Depositing Lab: Gregory Stephanopoulos
• Publication: Chatzivasileiou et al Proc Natl Acad Sci U S A. 2018 Dec 24. pii: 1812935116. doi: 10.1073/pnas.1812935116.

Backbone

• Vector backbone: psc101
• Backbone size w/o insert (bp): 6200
• Total vector size (bp): 11922
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Spectinomycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: GGPP synthase
• Alt name: GGPPs
• Species: Taxus canadensis
• Insert Size (bp): 891
• Mutation: codon optimized, truncated first 98 amino acids, methionine added. Please see Depositor Comments
• GenBank ID: AAD16018.1
• Promoter: T7

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: TTAATAAGGAGATATACCATATGTTCGACTTCAACGAG
• 3′ sequencing primer: TTGAACCCAAAAGGGCGGTATTAGTTTTGACGAAAGGC

Resource Information

• Supplemental Documents:
  • p5T7-LYCipi-ggpps.gb

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Based on pAC-LYCipi (Addgene plasmid # 53279 ; http://n2t.net/addgene:53279 ; RRID:Addgene_53279), with backbone changed to pSC101, and crtE swapped for ggpps from Taxus canadensis (truncated & codon optimized).

Please note: The plasmid contains a 387 nucleotide deletion from Trp249 of Eh-idi to His30 of IDI-2 (when compared to the depositor's sequence) combining the two ORFs into a single ORF. This mutation is not known to affect plasmid function.