pUC-ODN2
(Plasmid #208643)

• Purpose: (Empty Backbone) Empty vector for long single strand DNA (up to 1.5k) preparation. Further remove BsaI and BsmBI site in pUC-ODN (Addgene #208642).
• Depositing Lab: Quanfu Ma
• Publication: RBM24 interacts with RPL3 and regulates RPL3-eIF6 interaction (unpublished)

Backbone

• Vector backbone: pUC19
• Modifications to backbone: Remove BspQI (690), BsrDI (1760, 1934), BtsI (592, 2099, 2119) sites by point mutagenesis, remove BsaI (1760) site by point mutagenesis, remove BsmBI site by deletion of 2623~(origin)~85.
• Vector type: Long single strand DNA production

Growth in Bacteria

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

Resource Information

• Supplemental Documents:
  • pUC-ODN2.gb
• A portion of this plasmid was derived from a plasmid made by: point mutations that remove BspQI, BsrDI, BtsI were adoptted from Nagahora Hitoshi's work (genbank MA009870), point mutations that remove BsaI was adoptted from Wei Li's work (addgene#121957).

Terms and Licenses

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

Depositor Comments

Note that this is only a pUC19 removed indicated intrisic enzyme site, BspQI or BsrDI needed for ssDNA preparation need to be introduced together with the inserts. Method used was as discribed in "https://www.neb.com/-/media/nebus/files/application-notes/appnote_improved_methods_for_sdm_using_nebuilder_hifi_dna_assembly_master_mix.pdf"

>mutagenesis_primers

pUC-ΔBts592-F: 5' CGTTGCGCTCACAGCCCGCTTTCCAG 3'
pUC-ΔBts592-R: 5' CTGGAAAGCGGGCTGTGAGCGCAACG 3'

pUC-ΔBspQ690-F: 5' GTATTGGGCGCTGTTCCGCTTCCTC 3'
pUC-ΔBspQ690-R: 5' GAGGAAGCGGAACAGCGCCCAATAC 3'

pUC-ΔBsrD1760-F: 5' CAGTGCTGCAATAATACCGCGAGACC 3'
pUC-ΔBsrD1760-R: 5' GGTCTCGCGGTATTATTGCAGCACTG 3'

pUC-ΔBsrD1934-F: 5' CGTTGTTGCCATAGCTACAGGCATCGTG 3'
pUC-ΔBsrD1934-R: 5' CACGATGCCTGTAGCTATGGCAACAACG 3'

pUC-ΔBts2099&2119-F: 5' CCGCTGTGTTATCACTCATGGTTATGGCAGCACTACATAATTCTC 3'
pUC-ΔBts2099&2119-R: 5' TATGTAGTGCTGCCATAACCATGAGTGATAACACAGCGGCCAAC 3'

pUC-ΔBsa1766-F: 5' CAATAATACCGCGAGAGCCACGCTCACC 3'
pUC-ΔBsa1766-R: 5' GGTGAGCGTGGCTCTCGCGGTATTATTG 3'

pUC-Δ2623~85-F: 5' GAAAAGTGCCACCTGACGTCGAGCAGACAAGCCC 3'
pUC-Δ2623~85-R: 5' GGGCTTGTCTGCTCGACGTCAGGTGGCACTTTTC 3'