I am not an expert in this field, but I am very interested and have researched to find an answer. I received some assistance from tlooto.com for this response. Could you please review the response below to see if it is correct?

To design a pUCP18 vector from the pUC18 vector for cloning in Pseudomonas, incorporate a broad-host-range origin of replication such as oriV from plasmid RK2, ensuring compatibility with Pseudomonas species [2]. Use PCR to amplify the oriV region, then insert it into the pUC18 backbone via restriction enzyme digestion and ligation. Include antibiotic resistance markers suitable for selection in Pseudomonas [1]. Verify the construct through sequencing and test its functionality via transformation into Pseudomonas cells. This approach allows for stable replication and maintenance of the vector in Pseudomonas, facilitating successful gene cloning [3][4].

Reference

[1]

Matsusaki, H., Manji, S., Taguchi, K., Kato, M., Fukui, T., & Doi, Y. (1998). Cloning and Molecular Analysis of the Poly(3-hydroxybutyrate) and Poly(3-hydroxybutyrate-co-3-hydroxyalkanoate) Biosynthesis Genes in Pseudomonas sp. Strain 61-3. Journal of Bacteriology, 180, 6459 - 6467.

[2]

Aakvik, T., Degnes, K., Dahlsrud, R., Schmidt, F., Dam, R., Yu, L., Völker, U., Ellingsen, T., & Valla, S. (2009). A plasmid RK2-based broad-host-range cloning vector useful for transfer of metagenomic libraries to a variety of bacterial species.. FEMS microbiology letters, 296 2, 149-58 .

[3]

Yen, K., Karl, M., Blatt, L., Simon, M., Winter, R. B., Fausset, P., HSIENGS., L., Harcourt, A., Chen, K. K., & Amgen (1991). Cloning and characterization of a Pseudomonas mendocina KR1 gene cluster encoding toluene-4-monooxygenase. Journal of Bacteriology, 173, 5315 - 5327.

[4]

Kimbara, K., Hashimoto, T., Fukuda, M., Koana, T., Takagi, M., Oishi, M., & Yano, K. (1989). Cloning and sequencing of two tandem genes involved in degradation of 2,3-dihydroxybiphenyl to benzoic acid in the polychlorinated biphenyl-degrading soil bacterium Pseudomonas sp. strain KKS102. Journal of Bacteriology, 171, 2740 - 2747.