LOCUS pOME0190 683 bp DNA linear UNA 04-Mar-2014 DEFINITION MERS EMC nsp1 altered TAGGED; pDEST47 SOURCE UNC178-6 (MERS EMC nsp1 altered TAGGED) COMMENT RG altered to get rid of internal ORF in pOME185, 186 Plasmid synthesized by BioBasic Sequence verified via Sanger sequencing through Genewiz pOME= plasmid of the Orfeome project nsP= non-structural protein EMC= isolated in Netherlands FEATURES Location/Qualifiers Site join(1..6,1^2) /site_type="restriction site" /note="Name: SalI" /note="Pattern: gtcgac" /note="cut_0_on_positive_strand: 1^2" /note="cut_0_on_negative_strand: 5^6" /note="inhibited_by: 5': N6-methyladenosine" /note="star_activity: yes" /note="site_type: other" /note="restriction site" /label=SalI Site 1..12 /site_type="mutagenized site" /label="5' RE tag" Site join(7..12,11^12) /site_type="restriction site" /note="Name: KpnI" /note="Pattern: ggtacc" /note="cut_0_on_positive_strand: 11^12" /note="cut_0_on_negative_strand: 7^8" /note="inhibited_by: 5': N6-methyladenosine" /note="site_type: other" /note="restriction site" /label=KpnI Site 13..18 /site_type="mutagenized site" /label="Kozak sequence" CDS 19..666 /cds_type=ORF /note="Length: 648" /note="Found at strand: positive" /note="Start codon: ATG" Site 598..663 /site_type="mutagenized site" /label="3X FLAG tag" Site 664..669 /site_type="mutagenized site" /label="double stop" Site join(670..675,670^671) /site_type="restriction site" /note="Name: XhoI" /note="Pattern: ctcgag" /note="cut_0_on_positive_strand: 670^671" /note="cut_0_on_negative_strand: 674^675" /note="inhibited_by: 5': N6-methyladenosine" /note="site_type: other" /note="restriction site" /label=XhoI Site 670..683 /site_type="mutagenized site" /label="3' RE tag" Site join(676..683,677^678) /site_type="restriction site" /note="Name: NotI" /note="Pattern: gcggccgc" /note="cut_0_on_positive_strand: 677^678" /note="cut_0_on_negative_strand: 681^682" /note="inhibited_by: 5': N4-methylcytosine" /note="site_type: other" /note="restriction site" /label=NotI ORIGIN 1 gtcgacggta ccgccaccAT GTCTTTCGTG GCTGGTGTGA CCGCGCAAGG TGCGCGCGGT 61 ACGTATCGAG CAGCGCTCAA CTCTGAAAAA CATCAAGACC ATGTGTCTCT AACTGTGCCA 121 CTCTGTGGTT CAGGAAACCT GGTTGAAAAA CTTTCACCAT GGTTCATGGA TGGCGAAAAT 181 GCCTATGAAG TGGTGAAGGC CATGTTACTT AAAAAGGAGC CACTTCTCTA cGTGCCCATC 241 CGGCTGGCTG GACACACTAG ACACCTCCCA GGTCCTCGTG TGTACCTGGT TGAGAGGCTC 301 ATTGCTTGTG AAAATCCATT CATGGTTAAC CAATTGGCTT ATAGCTCTAG TGCAAAcGGC 361 AGCCTGGTTG GCACAACTTT GCAGGGCAAG CCTATTGGTA TGTTCTTCCC TTAcGACATC 421 GAACTTGTCA CAGGAAAGCA AAATATTCTC CTGCGCAAGT AcGGCCGTGG TGGTTATCAC 481 TACACCCCAT TCCACTAcGA GCGAGACAAC ACCTCTTGCC CTGAGTGGAT GGACGATTTT 541 GAGGCGGATC CTAAAGGCAA ATATGCCCAG AATCTGCTTA AGAAGTTGAT TGGCGGTgac 601 tacaaagacc atgacggtga ttataaagat catgatatcg attacaagga tgacgatgac 661 aagtagtaac tcgaggcggc cgc //