pSiMPlh_N + pSiMPlh_C
(Plasmid #134316)

• Purpose: Maintenance of 2 plasmids with a single antibiotic in E. coli
• Depositing Lab: Barbara Di Ventura
• Publication: Palanisamy et al Nat Commun. 2019 Oct 31;10(1):4967. doi: 10.1038/s41467-019-12911-1.

Backbone

• Vector backbone: pBAD33 + pTrc99a
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Hygromycin, 200 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: hygR_gp41-1 N-intein + hygR_gp41-1 C-intein
• Alt name: HPT
• Species: Synthetic
• Mutation: P254Q in hygR_gp41-1 N-intein

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: Unknown

Terms and Licenses

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

Depositor Comments

pSiMPlh_N should be cotransformed with pSiMPlh_C (or vice versa) in E. Coli to get a reconstituted functional HPT thereby conferring resistance against hygromycin. mRuby3 gene was amplified from a construct bought by us from Addgene (ID: 74252).

Please see the Sequences link for the individual plasmid sequences.

The sample from Addgene will have both the plasmids pSiMPlh_N + pSiMPlh_C in it for the interested
antibiotic resistance. Separating individual plasmid backbones will be possible via PCR. Agarose gel electrophoresis can be used to visualize the presence of both the plasmids. But, separating individual plasmids via gel electrophoresis, in our hands, always resulted in cross-contamination of the two backbones.