pRL128
(Plasmid #40180)

• Depositing Lab: Eric Cascales
• Publication: Gueguen et al Appl Environ Microbiol. 2013 Jan;79(1):32-8. doi: 10.1128/AEM.02504-12. Epub 2012 Oct 12.

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

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

Resource Information

• Articles Citing this Plasmid:
  • 3 References

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

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.