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Engineering a minimal cloning vector from a pUC18 plasmid backbone with an extended multiple cloning site

Jens Staal (UGent) , Kûbra Alci (UGent) , Wouter De Schamphelaire (UGent) , Martine Vanhoucke (UGent) and Rudi Beyaert (UGent)
(2019) BIOTECHNIQUES. 66(6). p.254-259
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Abstract
Minimal plasmids play an essential role in many intermediate steps in molecular biology. For example, they can be used to assemble building blocks in synthetic biology or be used as intermediate cloning plasmids that are ideal for PCR-based mutagenesis methods. A small backbone also opens up for additional unique restriction enzyme cloning sites. Here we describe the generation of pICOz, a 1185-bp fully functional high-copy cloning plasmid with an extended multiple cloning site. We believe that this is the smallest high-copy cloning vector ever described. METHOD SUMMARY: We eliminated all superfluous sequences in a commonly used cloning vector in order to generate as small a cloning plasmid as possible by simple iterative PCR mutagenesis.
Keywords
cloning, Escherichia coli, minimalism, mutagenesis, PCR, plasmids, recombination, repository, synthetic biology, synthetic nucleotide, DNA, REPLICATION, MUTAGENESIS, RESISTANCE

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Citation

Please use this url to cite or link to this publication:

MLA
Staal, Jens et al. “Engineering a Minimal Cloning Vector from a pUC18 Plasmid Backbone with an Extended Multiple Cloning Site.” BIOTECHNIQUES 66.6 (2019): 254–259. Print.
APA
Staal, J., Alci, K., De Schamphelaire, W., Vanhoucke, M., & Beyaert, R. (2019). Engineering a minimal cloning vector from a pUC18 plasmid backbone with an extended multiple cloning site. BIOTECHNIQUES, 66(6), 254–259.
Chicago author-date
Staal, Jens, Kûbra Alci, Wouter De Schamphelaire, Martine Vanhoucke, and Rudi Beyaert. 2019. “Engineering a Minimal Cloning Vector from a pUC18 Plasmid Backbone with an Extended Multiple Cloning Site.” Biotechniques 66 (6): 254–259.
Chicago author-date (all authors)
Staal, Jens, Kûbra Alci, Wouter De Schamphelaire, Martine Vanhoucke, and Rudi Beyaert. 2019. “Engineering a Minimal Cloning Vector from a pUC18 Plasmid Backbone with an Extended Multiple Cloning Site.” Biotechniques 66 (6): 254–259.
Vancouver
1.
Staal J, Alci K, De Schamphelaire W, Vanhoucke M, Beyaert R. Engineering a minimal cloning vector from a pUC18 plasmid backbone with an extended multiple cloning site. BIOTECHNIQUES. 2019;66(6):254–9.
IEEE
[1]
J. Staal, K. Alci, W. De Schamphelaire, M. Vanhoucke, and R. Beyaert, “Engineering a minimal cloning vector from a pUC18 plasmid backbone with an extended multiple cloning site,” BIOTECHNIQUES, vol. 66, no. 6, pp. 254–259, 2019.
@article{8619550,
  abstract     = {Minimal plasmids play an essential role in many intermediate steps in molecular biology. For example, they can be used to assemble building blocks in synthetic biology or be used as intermediate cloning plasmids that are ideal for PCR-based mutagenesis methods. A small backbone also opens up for additional unique restriction enzyme cloning sites. Here we describe the generation of pICOz, a 1185-bp fully functional high-copy cloning plasmid with an extended multiple cloning site. We believe that this is the smallest high-copy cloning vector ever described.
METHOD SUMMARY: We eliminated all superfluous sequences in a commonly used cloning vector in order to generate as small a cloning plasmid as possible by simple iterative PCR mutagenesis.},
  author       = {Staal, Jens and Alci, Kûbra and De Schamphelaire, Wouter and Vanhoucke, Martine and Beyaert, Rudi},
  issn         = {0736-6205},
  journal      = {BIOTECHNIQUES},
  keywords     = {cloning,Escherichia coli,minimalism,mutagenesis,PCR,plasmids,recombination,repository,synthetic biology,synthetic nucleotide,DNA,REPLICATION,MUTAGENESIS,RESISTANCE},
  language     = {eng},
  number       = {6},
  pages        = {254--259},
  title        = {Engineering a minimal cloning vector from a pUC18 plasmid backbone with an extended multiple cloning site},
  url          = {http://dx.doi.org/10.2144/btn-2019-0014},
  volume       = {66},
  year         = {2019},
}

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