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Addgene

p1A Prk-
(Plasmid #162694)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 162694 Standard format: Plasmid sent in bacteria as agar stab 1 $85
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Backbone

  • Vector backbone
    pFE21 (derived from pZE21expressing under PLtet0-1 and constitutive tetR)
  • Backbone size w/o insert (bp) 3127
  • Total vector size (bp) 5986
  • Vector type
    Bacterial Expression

Growth in Bacteria

  • Bacterial Resistance(s)
    Kanamycin, 50 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Growth instructions
    Do not induce with aTc in LB, it will induce prk toxicity. If you want to express the genes do so in minimal media.
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    H. neapolitanus CbbLS and S. elongatus Prk
  • Species
    H. neapolitanus and S. elongatus
  • Insert Size (bp)
    2859
  • Mutation
    Prk inactive (K20M S21A native protein, this map has a 6x N-terminal His tagged PRK so its K26M, S27A)
  • Promoter PLtet0-1 promoter
  • Tag / Fusion Protein
    • N terminal 6x His tag on PRK (N terminal on insert)

Cloning Information

  • Cloning method Gibson Cloning
  • 5′ sequencing primer gaaagccatccagtttactttgca
  • 3′ sequencing primer GCGTTCACCGACAAACAACA
    • (Common Sequencing Primers)

Resource Information

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry
Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

This plasmid contains a Prk K20M S21A mutation rendering Prk inactive.

How to cite this plasmid ( Back to top)

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:

    p1A Prk- was a gift from David Savage (Addgene plasmid # 162694 ; http://n2t.net/addgene:162694 ; RRID:Addgene_162694)

  • For your References section:

    Functional reconstitution of a bacterial CO2 concentrating mechanism in E. coli. Flamholz AI, Dugan E, Blikstad C, Gleizer S, Ben-Nissan R, Amram S, Antonovsky N, Ravishankar S, Noor E, Bar-Even A, Milo R, Savage D. Elife. 2020 Oct 21;9. pii: 59882. doi: 10.7554/eLife.59882. 10.7554/eLife.59882 PubMed 33084575
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