pRiboI-Sec-APEX2m
(Plasmid #176845)

• Purpose: Expression of APEX2 in the periplasm from a codon-optimized gene (single-copy integrating plasmid)
• Depositing Lab: Jessica Seeliger
• Publication: Ahamed et al Methods Enzymol. 2022;664:267-289. doi: 10.1016/bs.mie.2021.11.021. Epub 2021 Dec 30.

Backbone

• Vector backbone: pMV306
• Backbone manufacturer: Jeffery Cox Lab
• Backbone size w/o insert (bp): 3981
• Total vector size (bp): 5086
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: When the plasmid is used in mycobacteria, add theophylline to a maximum of 2mM to promote gene expression.
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: pRibo-Sec-APEX2m
• Alt name: pRibo aka.; hsp60 based theophylline (on) riboswitch promotor
• Alt name: Sec aka; Sec signal; secretion signal from Mpt63 mycobacterial protein
• Alt name: APEX2m aka.; ascorbate peroxidase 2 (codon optimized for mycobacteria)
• Insert Size (bp): 1183
• Promoter: pRibo, theophilline riboswitch inducible hsp60 promoter

Cloning Information

• Cloning method: Ligation Independent Cloning
• 5′ sequencing primer: GCCTTTGAGTGAGCTGATACC
• 3′ sequencing primer: GATGGCATAAAACGAAAGGCC

Resource Information

• Supplemental Documents:
  • pRiboI-Sec-APEX2m.gb
  • pRiboI-Sec-APEX2m.gb
• A portion of this plasmid was derived from a plasmid made by: mycobacterial codon optimized sequence of Apex2 from Alice Ting Lab. Ordered gene from Genewiz.

Terms and Licenses

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

Depositor Comments

Please note: Plasmid contains an E151V mutation in the int feature. This mutation is not known to affect plasmid function.