pXZ13lacZalpha
(Plasmid #160229)

• Purpose: (Empty Backbone) places LacZalpha unit between Esp3I sites for better cutting of plasmid and ability to distinguish recombinants as white colonies when plated on Xgal plates
• Depositing Lab: Ian Duncan
• Publication: Modified donor plasmids to enhance Golden Gate assembly of Drosophila two stage Crispr/Cas9 cassette exchange (unpublished)

Backbone

• Vector backbone: pXZ13 (Addgene plasmid #60949)
• Backbone manufacturer: Zhang et al. G3. 2014 Oct 15;4(12):2409-18
• Backbone size (bp): 2528
• Modifications to backbone: pXZ13 was digested with Esp3I and the lacZ alpha subunit from pEMBL18- was cloned in as a compatible PCR fragment
• Vector type: Bacterial Expression, CRISPR
• Promoter: lac

Growth in Bacteria

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

Cloning Information

• Cloning method: Gibson Cloning

Terms and Licenses

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

Depositor Comments

This vector is an improvement upon the published pXZ13 vector for Golden Gate cloning of the donor plasmid in the published article noted below. The vector increases cloning efficiency in two ways: 1) the distance between the Esp3I sites is increased to insure complete cutting at both sites, and 2) lacZ alpha is placed between the two cutting sites to provide Blue/White screening of recombinants.
Vector progenitor reference:
Zhang X, Koolhaas WH, Schnorrer F. A Versatile Two-Step CRISPR- and RMCE-Based Strategy for Efficient Genome Engineering in Drosophila. G3(Bethesda). 2014 Oct 15;4(12):2409-18.