pRecomb-loxP-mCherry-4Gcloningsite-lox2272-66
(Plasmid #168787)

• Purpose: (Empty Backbone) Donor vector for recombinase mediated integration in filamentous fungi. Is a shuttle vector for optional in vivo cloning in yeast, and contains four inducible promoters or sites for whole BGC cloning.
• Depositing Lab: Yit Heng Chooi
• Publication: Roux et al ACS Synth Biol. 2022 Feb 16. doi: 10.1021/acssynbio.1c00458.

Backbone

• Vector backbone: pRecomb-loxp-mCherry-4Gcloningsite-lox2272-66
• Modifications to backbone: pRecomb was created de novo by assembly of parts from pYFAC-CH2 and pKW20088, excluding AMA1 origin of replication.
• Vector type: Cre/Lox, Synthetic Biology ; Expression in filamentous fungi
• Selectable markers: URA3 ; pyrG

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: For improved transformation efficiency, using a DH10B E coli such as 10-beta, and electroporation.
• Copy number: High Copy

Cloning Information

• Cloning method: Gibson Cloning

Terms and Licenses

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

Depositor Comments

pRecomb-loxp-mcherry-4Gcloningsite-lox71-2272-66 (donor vector 2) is shuttle vector that supports optional transformation-associated recombination cloning in Saccharomyces cerevisiae. Donor vector 2 also contains within the floxed region four cloning sites for the expression of biosynthetic genes under strong alcohol inducible promoters, derived from the vector pYFAC-CH2. As the four-promoter multiple cloning site is flanked by EcoRI sites, donor vector 2 can also be used for cloning genomic fragments containing a BGC.