pdHJ4
(Plasmid #248895)

• Purpose: Generation of double Holliday Junction DNA
• Depositing Lab: Stephen West
• Publication: Ho et al Proc Natl Acad Sci U S A. 2022 May 3;119(18):e2123420119. doi: 10.1073/pnas.2123420119. Epub 2022 Apr 22.

Backbone

• Vector backbone: pBluescript II SK(+)
• Backbone manufacturer: Agilent
• Backbone size w/o insert (bp): 2464
• Total vector size (bp): 3486
• Modifications to backbone: pBluescript II SK(+) was digested with SapI and PvuII and synthetic DNA fragments were added as follows 1) 25- to 30-bp homology to pBluescript II SK(+) backbone (SapI site); 2) First DNAzyme self-cleavage cassette 3) precursor DNA 4) Second DNAzyme self-cleavage cassette 5) 25- to 30-bp homology to pBluescript II SK(+) backbone (PvuII site). Homologous sequences 1 and 5 allow the precursor DNA to be inserted with the intended orientation into the backbone during Gibson assembly reactions. Priginal SapI site near ori was lost in the resultant phagemids. The sequence of the precursor DNA (d4), was based on d1 (from pdHJ1), so that so that it could be used to generate DNA molecules containing Holliday junctions.
• Vector type: Cloning and Sequencing. As a phagemid, it allows for the rescue of single stranded DNA

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: Transforming pdHJ4 into E. coli DH5α together with an M13 helper plasmid yields single-stranded DNA.
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: synthetic DNA comprised of homologous sequences (to phagemid backbone), DNAzyme self-cleavage cassettes and precursor DNA
• Species: Synthetic
• Insert Size (bp): 1022
• GenBank ID:

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: n/a

Terms and Licenses

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