pSiMPlk_N + pSiMPlk_C
(Plasmid #134310)

• 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): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: kanR_gp41-1 N-intein + kanR_gp41-1 C-intein
• Alt name: APT
• Species: Synthetic

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

pSiMPlk_N should be cotransformed with pSiMPlk_C (or vice versa) in E. coli to get a reconstituted functional APT thereby conferring resistance against kanamycin. 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 pSiMPlk_N + pSiMPlk_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.