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pEP PolI
(Plasmid #11722)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 11722 Standard format: Plasmid sent in bacteria as agar stab 1 $75

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pHSG576
  • Backbone manufacturer
    see NCBI D88215
  • Backbone size w/o insert (bp) 3475
  • Vector type
    Bacterial Expression

Growth in Bacteria

  • Bacterial Resistance(s)
    Chloramphenicol
  • Growth Temperature
    30°C
  • Growth Strain(s)
    JS200
  • Growth instructions
    Grow at 30oC in LB in JS200 cells (polA recA718). The cells need to be kept in exponential phase (do not let the culture reach saturation)!
  • Copy number
    Low Copy

Gene/Insert

  • Gene/Insert name
    DNA polymerase I EP
  • Alt name
    DNA polymerase I
  • Alt name
    PolA
  • Alt name
    PolI
  • Species
    E. coli
  • Insert Size (bp)
    2787
  • Mutation
    D424A, I709N, A759R. Error-prone version of PolI.
  • Entrez Gene
    polA (a.k.a. b3863, ECK3855, JW3835, resA)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site HindIII (not destroyed)
  • 3′ cloning site EcoRI (not destroyed)
  • 5′ sequencing primer m13 reverse primer
  • 3′ sequencing primer m13 forward 20 primer
  • (Common Sequencing Primers)

Resource Information

Depositor Comments

See attached mutagenesis protocol provided by depositing scientist.

Please note that this plasmid may require a unique bacterial strain, so make sure to confirm that you can also obtain the appropriate growth strain. Please contact us at [email protected] or contact our distributors if you have any questions.

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:

    pEP PolI was a gift from Lawrence Loeb (Addgene plasmid # 11722 ; http://n2t.net/addgene:11722 ; RRID:Addgene_11722)
  • For your References section:

    Targeted gene evolution in Escherichia coli using a highly error-prone DNA polymerase I. Camps M, Naukkarinen J, Johnson BP, Loeb LA. Proc Natl Acad Sci U S A. 2003 Aug 19. 100(17):9727-32. 10.1073/pnas.1333928100 PubMed 12909725