LOCUS Exported 5682 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Yeast Expression plasmid for SARS-CoV-2 nsp6. ACCESSION . VERSION . KEYWORDS pGBW-m4046504 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5682) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 5682) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5682 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 25..466 /gene="S. cerevisiae GAL1" /label=GAL1 promoter /note="inducible promoter, regulated by Gal4" protein_bind 25..142 /label=UAS /bound_moiety="Gal4" /note="upstream activating sequence mediating Gal4-dependent induction" primer_bind 437..460 /label=GAL1 /note="S. cerevisiae GAL1 promoter, forward primer" CDS 1351..1368 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" CDS 1369..1392 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" primer_bind complement(1437..1455) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 1806..2609 /codon_start=1 /gene="S. cerevisiae URA3" /product="orotidine-5'-phosphate decarboxylase, required for uracil biosynthesis" /label=URA3 /note="yeast auxotrophic marker, counterselectable with 5-fluoroorotic acid (5-FOA)" /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN" rep_origin 2692..3674 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 3752..3781 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 3803..4390 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 4402..4429 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(4453..4644) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(4645..5505) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESLRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLATIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGSLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 5268..5287 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5506..5608) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 5609..5640 /label=tonB terminator /note="bidirectional E. coli tonB-P14 transcription terminator" ORIGIN 1 gcggccgctt gctatattga agtacggatt agaagccgcc gagcgggcga cagccctccg 61 acggaagact ctcctccgtg cgtcctcgtc ttcaccggtc gcgttcctga aacgcagatg 121 tgcctcgcgc cgcactgctc cgaacaataa agattctaca atactagctt ttatggttat 181 gaagaggaaa aattggcagt aacctggccc cacaaacctt caaattaacg aatcaaatta 241 acaaccatag gatgataatg cgattagttt tttagcctta tttctggggt aattaatcag 301 cgaagcgatg atttttgatc tattaacaga tatataaatg gaaaagctgc ataaccactt 361 taactaatac tttcaacatt ttcagtttgt attacttctt attcaaatgt cataaaagta 421 tcaacaaaaa attgttaata tacctctata ctttaacgtc aaggagaaaa aactataatg 481 tctgctgtca aaagaactat taagggtacc catcactggt tgctgctaac aatcttaact 541 tccttgttag tgcttgttca aagtacccag tggtcgttgt ttttcttctt gtacgagaac 601 gcctttctcc cattcgcaat ggggataatt gctatgtcag cttttgccat gatgttcgtt 661 aagcacaaac atgcgttttt gtgcttattc ttgttgcctt ctctcgctac ggtagcttat 721 ttcaatatgg tctacatgcc agcttcctgg gttatgcgta tcatgacttg gttggacatg 781 gttgatacca gcctttctgg tttcaagttg aaagattgtg tcatgtacgc atctgccgtt 841 gtattactaa ttcttatgac tgctagaacc gtctatgacg atggcgcgag aagggtttgg 901 acactaatga acgtgttgac tttagtttac aaggtctact atggaaatgc cttggaccaa 961 gctatctcca tgtgggcttt aataatctca gtcacttcta actacagtgg tgttgttact 1021 acagtgatgt ttttggctag aggtattgtc ttcatgtgtg ttgaatattg tcccattttc 1081 tttatcaccg gtaacacttt acaatgcatt atgttggttt actgtttctt aggttacttt 1141 tgtacctgtt atttcggcct attttgtttg ttgaatcgat actttagact cacattgggt 1201 gtttacgatt atttggtcag cactcaagaa tttagatata tgaactccca aggtttgtta 1261 ccaccaaaga actctattga tgccttcaag ttgaacatca agctgttagg tgtcggtgga 1321 aaaccttgca tcaaggtagc caccgtccaa ctcgtgccga gaggttcttg gagccatccg 1381 cagtttgaaa aataaacagg ccccttttcc tttgtcgata tcatgtaatt agttatgtca 1441 cgcttacatt cacgccctcc tcccacatcc gctctaaccg aaaaggaagg agttagacaa 1501 cctgaagtct aggtccctat ttattttttt taatagttat gttagtatta agaacgttat 1561 ttatatttca aatttttctt ttttttctgt acaaagcggc cgccgaacca ctagtgcact 1621 gcagtacacg gtttccttga aatttttttg attcggtaat ctccgaacag aaggaagaac 1681 gaaggaagga gcacagactt agattggtat atatacgcat atgtagtgtt gaagaaacat 1741 gaaattgccc agtattctta acccaactgc acagaacaaa aacgtgcagg aaacgaagat 1801 aaatcatgtc gaaagctaca tataaggaac gtgctgctac tcatcctagt cctgttgctg 1861 ccaagctatt taatatcatg cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc 1921 gtaccaccaa ggaattactg gagttagttg aagcattagg tcccaaaatt tgtttactaa 1981 aaacacatgt ggatatcttg actgattttt ccatggaggg cacagttaag ccgctaaagg 2041 cattatccgc caagtacaat tttttactct tcgaggacag aaaatttgct gacattggta 2101 atacagtcaa attgcagtac tctgcgggtg tatacagaat agcagaatgg gcagacatta 2161 cgaatgcaca cggtgtggtg ggcccaggta ttgttagcgg tttgaagcag gcggcagaag 2221 aagtaacaaa ggaacctaga ggccttttga tgttagcaga attgtcatgc aagggctccc 2281 tatctactgg agaatatact aagggtactg ttgacattgc gaagagcgac aaagattttg 2341 ttatcggctt tattgctcaa agagacatgg gtggaagaga tgaaggttac gattggttga 2401 ttatgacacc cggtgtgggt ttagatgaca agggagatgc attgggtcaa cagtatagaa 2461 ccgtggatga tgtggtttct acaggatctg acattattat tgttggaaga ggactatttg 2521 caaagggaag ggatgctaag gtagagggtg aacgttacag aaaagcaggc tgggaagcat 2581 atttgagaag atgcggccag caaaactaaa aaactgtatt ataagtaaat gcatgtatac 2641 taaactcaca aattagagct tcaatttaat tatatcagtt attacccgag taacgaagca 2701 tctgtgcttc attttgtaga acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag 2761 aatctgagct gcatttttac agaacagaaa tgcaacgcga aagcgctatt ttaccaacga 2821 agaatctgtg cttcattttt gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa 2881 caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgagagcg ctattttacc 2941 aacaaagaat ctatacttct tttttgttct acaaaaatgc atcccgagag cgctattttt 3001 ctaacaaagc atcttagatt actttttttc tcctttgtgc gctctataat gcagtctctt 3061 gataactttt tgcactgtag gtccgttaag gttagaagaa ggctactttg gtgtctattt 3121 tctcttccat aaaaaaagcc tgactccact tcccgcgttt actgattact agcgaagctg 3181 cgggtgcatt ttttcaagat aaaggcatcc ccgattatat tctataccga tgtggattgc 3241 gcatactttg tgaacagaaa gtgatagcgt tgatgattct tcattggtca gaaaattatg 3301 aacggtttct tctattttgt ctctatatac tacgtatagg aaatgtttac attttcgtat 3361 tgttttcgat tcactctatg aatagttctt actacaattt ttttgtctaa agagtaatac 3421 tagagataaa cataaaaaat gtagaggtcg agtttagatg caagttcaag gagcgaaagg 3481 tggatgggta ggttatatag ggatatagca cagagatata tagcaaagag atacttttga 3541 gcaatgtttg tggaagcggt attcgcaata ttttagtagc tcgttacagt ccggtgcgtt 3601 tttggttttt tgaaagtgcg tcatcagagc gcttttggtt ttcaaaagcg ctctgaagtt 3661 cctatacttt ctagctagag aataggaact tcccgagcgg ccgcgtgtta caagcttgac 3721 tccagcgtaa ctggactgca atcaactcac tggctcacct tcacgggtgg gcctttcttc 3781 ggtagaaaat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc 3841 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc 3901 tttttccgag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta 3961 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct 4021 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc 4081 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca 4141 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga 4201 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg 4261 aacaggagag cgcacgaggg agcttccagg ggtaaacgcc tggtatcttt atagtcctgt 4321 cgggtttcgc cacctctgac ttgagcatcg atttttgtga tgctcgtcag gggggcggag 4381 cctatggaaa aacgccagca acgcagaaag gcccacccga aggtgagcca ggtgattaca 4441 tttgggccct catcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta 4501 aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat ccgcgtccag 4561 ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag 4621 ggcggttttt tcctgtttgg tcacttacca atgcttaatc agtgaggcac ctatctcagc 4681 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 4741 gcgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 4801 ggctccagat ttatcagcaa taaaccagcc agccgggagg gccgagcgca gaagtgatcc 4861 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 4921 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 4981 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 5041 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgccagaag 5101 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 5161 catgccatcc gtgagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 5221 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 5281 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gtaaactctc 5341 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 5401 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 5461 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcattttag cttccttagc 5521 tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg 5581 aaagttggaa cctcttacgt gccgatcaag tcaaaagcct ccggtcggag gcttttgact 5641 ttctgctatg gaggtcaggt atgatttaaa tggtcagtta ga //