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pRL128
(Plasmid #40180)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 40180 Standard format: Plasmid sent in bacteria as agar stab 1 $85

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pKD13
  • Backbone size w/o insert (bp) 3434
  • Total vector size (bp) 4922
  • Modifications to backbone
    The pKD13-derived pRL128 plasmid was constructed in two steps by restriction-free (RF) cloning using oligonucleotides with a 3’ target-specific sequence and a 5’ extension complementary to the desired insertion site in the pKD13 destination plasmid. First, the Ptac-lacO DNA region of the pMAL-p2 plasmid was amplified by PCR. The PCR product was then used as oligonucleotides for a second PCR using pKD13 as a template. The resulting plasmid, carrying the Ptac-lacO DNA fragment downstream the FRT site and KanR-encoding gene, was named pRL127. Then, the araC-PBAD DNA region of pBAD18 was amplified by PCR. The PCR product was used as oligonucleotides for a second PCR using pRL127 as a template. In the resulting plasmid designated pRL128, the Ptac and PBAD promoters are in divergent orientation and flank the two FRT sites.

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin and Kanamycin, 100 & 50 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    CC118lambdapir
  • Copy number
    Low Copy

Gene/Insert 1

  • Gene/Insert name
    Ptac-lacO
  • Insert Size (bp)
    290
  • Promoter tac

Cloning Information for Gene/Insert 1

  • Cloning method Ligation Independent Cloning
  • 5′ sequencing primer pKD13-ptac#1
  • 3′ sequencing primer pKD13-ptac#2
  • (Common Sequencing Primers)

Gene/Insert 2

  • Gene/Insert name
    araC-pBAD
  • Insert Size (bp)
    1198
  • Promoter BAD

Cloning Information for Gene/Insert 2

  • Cloning method Ligation Independent Cloning
  • 5′ sequencing primer pKD13-pbad#1
  • 3′ sequencing primer pKD13-pbad#2
  • (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

Note that though there are some discrepancies between the Addgene quality control sequence and the depositor sequence, they should not affect the function of the plasmid.

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:

    pRL128 was a gift from Eric Cascales (Addgene plasmid # 40180 ; http://n2t.net/addgene:40180 ; RRID:Addgene_40180)
  • For your References section:

    Promoter swapping unveils the role of the Citrobacter rodentium CTS1 type VI secretion system in interbacterial competition. Gueguen E, Cascales E. Appl Environ Microbiol. 2013 Jan;79(1):32-8. doi: 10.1128/AEM.02504-12. Epub 2012 Oct 12. 10.1128/AEM.02504-12 PubMed 23064344