LOCUS Exported 6024 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Yeast Expression plasmid for SARS-CoV-2 N (nucleocapsid). ACCESSION . VERSION . KEYWORDS pGBW-m4046671 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6024) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 6024) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6024 /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 complement(667..678) /codon_start=1 /product="WELQut protease recognition and cleavage site" /label=WELQut site /translation="WELQ" primer_bind complement(1779..1797) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 2148..2951 /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 3034..4016 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 4094..4123 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 4145..4732 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 4744..4771 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(4795..4986) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(4987..5847) /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 5610..5629 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5848..5950) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 5951..5982 /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 tctgataatg gcccacagaa ccaacgtaac gcacctagaa tcaccttcgg tggtccatcc 541 gacagcactg gttcaaatca aaacggagag agatcgggtg cccggagtaa gcaaaggcgc 601 ccccaaggtc taccgaataa cacagcttct tggtttactg ctcttacgca acacggtaaa 661 gaagacttga agttcccaag aggtcaaggg gttccaatta acaccaactc ctctccagat 721 gatcaaatag gttactatag aagagctact agacgtatta ggggtgggga cggcaaaatg 781 aaggatttat ctcctagatg gtacttttat tacttgggta ccggtccaga agctggattg 841 ccttacggtg ccaataagga cggtatcatt tgggtagcaa ctgaaggtgc tttaaatacc 901 ccaaaggacc atatcggtac tagaaaccct gctaataacg ctgccattgt cttgcaatta 961 ccacagggga caacgttgcc aaagggtttc tacgccgaag gatccagagg tggctcgcaa 1021 gcttctagta gatcatcctc tcgcagcaga aactcatctc gtaactccac accaggtagt 1081 tcgagaggta cctctccagc gagaatggct ggtaacggtg gtgatgcagc tctggcgttg 1141 ctgctactcg acagattgaa tcaattggaa tccaaaatgt ctggtaaggg tcaacaacaa 1201 cagggtcaga ctgtgactaa gaagtctgct gctgaagctt ccaaaaagcc ccgtcaaaag 1261 agaaccgcca ccaaggccta taatgtcact caagctttcg gaagaagagg tccagagcaa 1321 acgcaaggta actttggaga tcaagaattg atccggcagg gtaccgatta taaacactgg 1381 ccccaaattg ctcaattcgc gccaagcgcc tcagcttttt tcggcatgtc aagaataggt 1441 atggaagtta ctccatctgg gacttggtta acttacactg gtgcaatcaa gttggacgat 1501 aaagacccaa attttaaaga ccaagttatt ctactcaaca agcatataga tgcttacaaa 1561 accttcccgc ctacagaacc caagaaagat aaaaagaaga aggctgacga gacacaagca 1621 ttgcctcagc gtcaaaagaa acaacaaact gtcaccttgt taccagcagc tgatcttgat 1681 gatttctcta agcaattgca acaatccatg tcgtccgccg actctactca agcgtaaaca 1741 ggcccctttt cctttgtcga tatcatgtaa ttagttatgt cacgcttaca ttcacgccct 1801 cctcccacat ccgctctaac cgaaaaggaa ggagttagac aacctgaagt ctaggtccct 1861 atttattttt tttaatagtt atgttagtat taagaacgtt atttatattt caaatttttc 1921 ttttttttct gtacaaagcg gccgccgaac cactagtgca ctgcagtaca cggtttcctt 1981 gaaatttttt tgattcggta atctccgaac agaaggaaga acgaaggaag gagcacagac 2041 ttagattggt atatatacgc atatgtagtg ttgaagaaac atgaaattgc ccagtattct 2101 taacccaact gcacagaaca aaaacgtgca ggaaacgaag ataaatcatg tcgaaagcta 2161 catataagga acgtgctgct actcatccta gtcctgttgc tgccaagcta tttaatatca 2221 tgcacgaaaa gcaaacaaac ttgtgtgctt cattggatgt tcgtaccacc aaggaattac 2281 tggagttagt tgaagcatta ggtcccaaaa tttgtttact aaaaacacat gtggatatct 2341 tgactgattt ttccatggag ggcacagtta agccgctaaa ggcattatcc gccaagtaca 2401 attttttact cttcgaggac agaaaatttg ctgacattgg taatacagtc aaattgcagt 2461 actctgcggg tgtatacaga atagcagaat gggcagacat tacgaatgca cacggtgtgg 2521 tgggcccagg tattgttagc ggtttgaagc aggcggcaga agaagtaaca aaggaaccta 2581 gaggcctttt gatgttagca gaattgtcat gcaagggctc cctatctact ggagaatata 2641 ctaagggtac tgttgacatt gcgaagagcg acaaagattt tgttatcggc tttattgctc 2701 aaagagacat gggtggaaga gatgaaggtt acgattggtt gattatgaca cccggtgtgg 2761 gtttagatga caagggagat gcattgggtc aacagtatag aaccgtggat gatgtggttt 2821 ctacaggatc tgacattatt attgttggaa gaggactatt tgcaaaggga agggatgcta 2881 aggtagaggg tgaacgttac agaaaagcag gctgggaagc atatttgaga agatgcggcc 2941 agcaaaacta aaaaactgta ttataagtaa atgcatgtat actaaactca caaattagag 3001 cttcaattta attatatcag ttattacccg agtaacgaag catctgtgct tcattttgta 3061 gaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt 3121 acagaacaga aatgcaacgc gaaagcgcta ttttaccaac gaagaatctg tgcttcattt 3181 ttgtaaaaca aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca 3241 tttttacaga acagaaatgc aacgcgagag cgctatttta ccaacaaaga atctatactt 3301 cttttttgtt ctacaaaaat gcatcccgag agcgctattt ttctaacaaa gcatcttaga 3361 ttactttttt tctcctttgt gcgctctata atgcagtctc ttgataactt tttgcactgt 3421 aggtccgtta aggttagaag aaggctactt tggtgtctat tttctcttcc ataaaaaaag 3481 cctgactcca cttcccgcgt ttactgatta ctagcgaagc tgcgggtgca ttttttcaag 3541 ataaaggcat ccccgattat attctatacc gatgtggatt gcgcatactt tgtgaacaga 3601 aagtgatagc gttgatgatt cttcattggt cagaaaatta tgaacggttt cttctatttt 3661 gtctctatat actacgtata ggaaatgttt acattttcgt attgttttcg attcactcta 3721 tgaatagttc ttactacaat ttttttgtct aaagagtaat actagagata aacataaaaa 3781 atgtagaggt cgagtttaga tgcaagttca aggagcgaaa ggtggatggg taggttatat 3841 agggatatag cacagagata tatagcaaag agatactttt gagcaatgtt tgtggaagcg 3901 gtattcgcaa tattttagta gctcgttaca gtccggtgcg tttttggttt tttgaaagtg 3961 cgtcatcaga gcgcttttgg ttttcaaaag cgctctgaag ttcctatact ttctagctag 4021 agaataggaa cttcccgagc ggccgcgtgt tacaagcttg actccagcgt aactggactg 4081 caatcaactc actggctcac cttcacgggt gggcctttct tcggtagaaa atcaaaggat 4141 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc 4201 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aggtaactgg 4261 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca 4321 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc 4381 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga 4441 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac 4501 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga 4561 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag 4621 ggagcttcca ggggtaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg 4681 acttgagcat cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag 4741 caacgcagaa aggcccaccc gaaggtgagc caggtgatta catttgggcc ctcatcagag 4801 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc 4861 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag 4921 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt 4981 ggtcacttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 5041 atccatagtt gcctgactcc ccgtcgtgta gataactacg atgcgggagg gcttaccatc 5101 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 5161 aataaaccag ccagccggga gggccgagcg cagaagtgat cctgcaactt tatccgcctc 5221 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 5281 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 5341 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 5401 aaaagcggtt agctccttcg gtcctccgat cgttgccaga agtaagttgg ccgcagtgtt 5461 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtgagatg 5521 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 5581 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 5641 agtgctcatc attggaaaac gttcttcggg gcgtaaactc tcaaggatct taccgctgtt 5701 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 5761 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 5821 ggcgacacgg aaatgttgaa tactcatttt agcttcctta gctcctgaaa atctcgataa 5881 ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg aacctcttac 5941 gtgccgatca agtcaaaagc ctccggtcgg aggcttttga ctttctgcta tggaggtcag 6001 gtatgattta aatggtcagt taga //