Preprint Article Version 2 This version is not peer-reviewed

Engineering a Minimal Cloning Vector from a pUC18 Plasmid Backbone with an Extended Multiple Cloning Site

Version 1 : Received: 16 July 2018 / Approved: 16 July 2018 / Online: 16 July 2018 (12:52:38 CEST)
Version 2 : Received: 12 October 2018 / Approved: 15 October 2018 / Online: 15 October 2018 (09:22:22 CEST)

A peer-reviewed article of this Preprint also exists.

Journal reference: Biotechniques 2019
DOI: 10.2144/btn-2019-0014

Abstract

Minimal plasmids play an essential role in many intermediate steps in molecular biology. They can for example 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 (MCS). To our knowledge, this is the smallest high-copy cloning vector ever described.

Subject Areas

PCR; recombination; cloning; engineering; biotechnology; synthetic biology; synthetic nucleotide; plasmids; repository; minimalism; Escherichia coli; mutagenesis

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