Nature - USA (2020-06-25)

1 nature research | reporting summary October 2018 Corresponding author(s): Ke Lan Last updated by author(s):Apr 2, 2020 Reporti ...

2 nature research | reporting summary October 2018 For a reference copy of the document with all sections, see nature.com/docume ...

Nature | Vol 582 | 25 June 2020 | 561 Article Rapid reconstruction of SARS-CoV-2 using a synthetic genomics platform Tran Thi Nh ...

562 | Nature | Vol 582 | 25 June 2020 Article the large genome size (27–31 kb) and occasional instability of cloned DNA in E. co ...

Nature | Vol 582 | 25 June 2020 | 563 and overlapping sequences with the TAR plasmid pVC604. To muta- genize the MERS-CoV clone, ...

564 | Nature | Vol 582 | 25 June 2020 Article First cases identied in Wuhan Early Dec 14 synSARS-CoV-2 DNA segments ordered 14 ...

Nature | Vol 582 | 25 June 2020 | 565 1.1, 2.2, and 3.1, plaques were readily detectable, demonstrating that infectious virus ha ...

Article Methods Cells and general culture conditions Vero, Vero B4 and Vero B6 cells (all ATCC) were cultured in Dulbecco’s modi ...

A single colony was grown in 20 ml of SD−His liquid medium, 1 ml ali- quots were removed and expanded in fresh medium every 12 h ...

Article v.2019.2.3. Results from virus growth kinetics were analysed and graphi- cally presented using GraphPad Prism v.8.3.0 fo ...

Raj, V. S. et al. Dipeptidyl peptidase 4 is a functional receptor for the emerging human coronavirus-EMC. Nature 495 , 251–254 ...

Article rMERS-CoVMock rMERS-CoV-GFP Mock b c a kb^0102030 S 3 GFP P2A 4a 4b E 5M 1a 1b S 3 4a 4b 5 E M N F2 F1 F4 F3 F5 F6 F7 F8 ...

A A AA AA A A A A A A AAA C C CC C C C C C C CC C C G G G G G G G G G G G G U U U UUU U U U U U UU U U U U U AAAU A U U U U U A ...

Article Extended Data Fig. 2 | Sequence analyses of the 5′ UTR of the SARS-CoV-2 genome. a, Sequence analysis using 5′R ACE. Res ...

Design Construct 1 Construct 2 Construct 3 Construct 4 Construct 5 Construct 6 rSARS-CoV-2 rSARS-CoV-2-GFP TAR cloning Positive ...

Article 1a 1b 7a S E 6 N kb 0102030 F5 F9 F4 F6 F12 F3 F2 F1 F10 F7 F8 F11 pT7-5’ UTR 3’ UTR-An a 3a/bM 7b 8 3a/b E M 6 7a GFP 7 ...

Extended Data Fig. 4 | Reconstruction of synSARS-CoV-2-GFP and TAR cloning of full-length synthetic fragments 5 and 7 in yeast. ...

Article Extended Data Fig. 5 | See next page for caption. ...

Extended Data Fig. 5 | Characterization of rSARS-CoV-2, rSARS-CoV-2-GFP and synSARS-CoV-2-GFP. a, Serum neutralization assay sho ...

Article Extended Data Table 1 | List of restriction enzymes used to linearize the YAC DNA for each virus before in vitro transcr ...