pDL1
(Plasmid #182263)

• Purpose: (Empty Backbone) Gibson cloning vector for synthesis of single and double stranded RNA
• Depositing Lab: Daniel Lobo
• Publication: Wolff et al STAR Protoc. 2022 Jun 14;3(3):101458. doi: 10.1016/j.xpro.2022.101458. eCollection 2022 Sep 16.

Backbone

• Vector backbone: pDL1
• Backbone manufacturer: Daniel Lobo
• Backbone size (bp): 2319
• Vector type: Gibson cloning vector for synthesis of single and double stranded RNA
• Promoter: T3 and SP6 for sense and antisense riboprobe synthesis, respectively, and T7 for dsRNA synthesis

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: M13 Forward
• 3′ sequencing primer: M13 Reverse

Resource Information

• Supplemental Documents:
  • pDL1.gb

Terms and Licenses

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

Depositor Comments

pDL1 includes T3 and SP6 promoters for single-stranded RNA synthesis, and two T7 promoters and terminators for dsRNA synthesis.

To insert the gene of interest, simultaneously restriction digest the vector with BspQI and Gibson assembly with the insert extended with the following Gibson overhangs:

Forward primer extended with 5’-TTAACCCTCACTAAAGGGAG-3’.
Reverse primer extended with 5’-GGGATTTAGGTGACACTATAGAA-3’.

After assembly, directly transform DH5-alpha cells. M13 forward and reverse universal sequences can be used for complete forward and reverse sequencing of the inserted gene.