Author: João Pedro Fonseca; Alain R. Bonny; G. Renuka Kumar; Andrew H. Ng; Jason Town; Qiu Chang Wu; Elham Aslankoohi; Susan Y. Chen; Patrick Harrigan; Lindsey C. Osimiri; Amy L. Kistler; Hana El-Samad
Title: A Toolkit for Rapid Modular Construction of Biological Circuits in Mammalian Cells Document date: 2018_12_26
ID: 1kugu5zk_9
Snippet: Within these defined parts, further specialization is easily achieved. For example, Part 3 and Part 4 vectors can be replaced with constituent a and b vectors where Part 3 can be replaced with Part 3a and 3b, and still connect to a Part 4. This allows combinations that can implement tethering of localization tags, protein domains or any desired coding sequence both N-and C-terminally with an innocuous linker sequence in between (Fig. 1b) . Additi.....
Document: Within these defined parts, further specialization is easily achieved. For example, Part 3 and Part 4 vectors can be replaced with constituent a and b vectors where Part 3 can be replaced with Part 3a and 3b, and still connect to a Part 4. This allows combinations that can implement tethering of localization tags, protein domains or any desired coding sequence both N-and C-terminally with an innocuous linker sequence in between (Fig. 1b) . Additionally, we have developed coupled Part 234 vectors to accommodate rapid cloning of small guide RNA (sgRNA) expression by oligo annealing for CRISPR/Cas9-related genetic constructs. Parts 6, 7 and 8 generally flank a typical TU and can encode the method of delivery to cells. For example, it can encode the homology arms for a locus of integration, or bacterial origin of replication and selection markers.
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