pSiMPlk_N + pSiMPlk_C
PurposeMaintenance of 2 plasmids with a single antibiotic in E. coli
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||134310||Standard format: Plasmid sent in bacteria as agar stab||1||$85|
This material is available to academics and nonprofits only.
Vector backbonepBAD33 + pTrc99a
Vector typeBacterial Expression
Growth in Bacteria
Bacterial Resistance(s)Kanamycin, 50 μg/mL
Copy numberHigh Copy
Gene/Insert namekanR_gp41-1 N-intein + kanR_gp41-1 C-intein
- Cloning method Gibson Cloning
- 5′ sequencing primer Unknown (Common Sequencing Primers)
Terms and Licenses
- Not Available to Industry
- Zeocin® is an InvivoGen trademark.
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.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:pSiMPlk_N + pSiMPlk_C was a gift from Barbara Di Ventura (Addgene plasmid # 134310 ; http://n2t.net/addgene:134310 ; RRID:Addgene_134310)
For your References section:Split intein-mediated selection of cells containing two plasmids using a single antibiotic. Palanisamy N, Degen A, Morath A, Ballestin Ballestin J, Juraske C, Ozturk MA, Sprenger GA, Youn JW, Schamel WW, Di Ventura B. Nat Commun. 2019 Oct 31;10(1):4967. doi: 10.1038/s41467-019-12911-1. 10.1038/s41467-019-12911-1 PubMed 31672972