LOCUS Exported 6019 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Yeast Expression plasmid for SARS-CoV-2 N (nucleocapsid). ACCESSION . VERSION . KEYWORDS pGBW-m4046539 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6019) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 6019) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6019 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 10..417 /gene="S. cerevisiae TEF1" /label=TEF1 promoter /note="promoter for EF-1-alpha" CDS complement(620..631) /codon_start=1 /product="WELQut protease recognition and cleavage site" /label=WELQut site /translation="WELQ" CDS 1688..1705 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" CDS 1706..1729 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" primer_bind complement(1774..1792) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 2143..2946 /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 3029..4011 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 4089..4118 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 4140..4727 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 4739..4766 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(4790..4981) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(4982..5842) /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 5605..5624 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5843..5945) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 5946..5977 /label=tonB terminator /note="bidirectional E. coli tonB-P14 transcription terminator" ORIGIN 1 gcggccgccc cacacaccat agcttcaaaa tgtttctact ccttttttac tcttccagat 61 tttctcggac tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag catactaaat 121 ttcccctctt tcttcctcta gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa 181 aaaagagacc gcctcgtttc tttttcttcg tcgaaaaagg caataaaaat ttttatcacg 241 tttctttttc ttgaaaattt ttttttttga tttttttctc tttcgatgac ctcccattga 301 tatttaagtt aataaacggt cttcaatttc tcaagtttca gtttcatttt tcttgttcta 361 ttacaacttt ttttacttct tgctcattag aaagaaagca tagcaatcta atctaagttt 421 taattacaaa atgtctgata atggcccaca gaaccaacgt aacgcaccta gaatcacctt 481 cggtggtcca tccgacagca ctggttcaaa tcaaaacgga gagagatcgg gtgcccggag 541 taagcaaagg cgcccccaag gtctaccgaa taacacagct tcttggttta ctgctcttac 601 gcaacacggt aaagaagact tgaagttccc aagaggtcaa ggggttccaa ttaacaccaa 661 ctcctctcca gatgatcaaa taggttacta tagaagagct actagacgta ttaggggtgg 721 ggacggcaaa atgaaggatt tatctcctag atggtacttt tattacttgg gtaccggtcc 781 agaagctgga ttgccttacg gtgccaataa ggacggtatc atttgggtag caactgaagg 841 tgctttaaat accccaaagg accatatcgg tactagaaac cctgctaata acgctgccat 901 tgtcttgcaa ttaccacagg ggacaacgtt gccaaagggt ttctacgccg aaggatccag 961 aggtggctcg caagcttcta gtagatcatc ctctcgcagc agaaactcat ctcgtaactc 1021 cacaccaggt agttcgagag gtacctctcc agcgagaatg gctggtaacg gtggtgatgc 1081 agctctggcg ttgctgctac tcgacagatt gaatcaattg gaatccaaaa tgtctggtaa 1141 gggtcaacaa caacagggtc agactgtgac taagaagtct gctgctgaag cttccaaaaa 1201 gccccgtcaa aagagaaccg ccaccaaggc ctataatgtc actcaagctt tcggaagaag 1261 aggtccagag caaacgcaag gtaactttgg agatcaagaa ttgatccggc agggtaccga 1321 ttataaacac tggccccaaa ttgctcaatt cgcgccaagc gcctcagctt ttttcggcat 1381 gtcaagaata ggtatggaag ttactccatc tgggacttgg ttaacttaca ctggtgcaat 1441 caagttggac gataaagacc caaattttaa agaccaagtt attctactca acaagcatat 1501 agatgcttac aaaaccttcc cgcctacaga acccaagaaa gataaaaaga agaaggctga 1561 cgagacacaa gcattgcctc agcgtcaaaa gaaacaacaa actgtcacct tgttaccagc 1621 agctgatctt gatgatttct ctaagcaatt gcaacaatcc atgtcgtccg ccgactctac 1681 tcaagcgctc gtgccgagag gttcttggag ccatccgcag tttgaaaaat aaacaggccc 1741 cttttccttt gtcgatatca tgtaattagt tatgtcacgc ttacattcac gccctcctcc 1801 cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta 1861 ttttttttaa tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt 1921 tttctgtaca aagcggccgc cgaaccacta gtgcactgca gtacacggtt tccttgaaat 1981 ttttttgatt cggtaatctc cgaacagaag gaagaacgaa ggaaggagca cagacttaga 2041 ttggtatata tacgcatatg tagtgttgaa gaaacatgaa attgcccagt attcttaacc 2101 caactgcaca gaacaaaaac gtgcaggaaa cgaagataaa tcatgtcgaa agctacatat 2161 aaggaacgtg ctgctactca tcctagtcct gttgctgcca agctatttaa tatcatgcac 2221 gaaaagcaaa caaacttgtg tgcttcattg gatgttcgta ccaccaagga attactggag 2281 ttagttgaag cattaggtcc caaaatttgt ttactaaaaa cacatgtgga tatcttgact 2341 gatttttcca tggagggcac agttaagccg ctaaaggcat tatccgccaa gtacaatttt 2401 ttactcttcg aggacagaaa atttgctgac attggtaata cagtcaaatt gcagtactct 2461 gcgggtgtat acagaatagc agaatgggca gacattacga atgcacacgg tgtggtgggc 2521 ccaggtattg ttagcggttt gaagcaggcg gcagaagaag taacaaagga acctagaggc 2581 cttttgatgt tagcagaatt gtcatgcaag ggctccctat ctactggaga atatactaag 2641 ggtactgttg acattgcgaa gagcgacaaa gattttgtta tcggctttat tgctcaaaga 2701 gacatgggtg gaagagatga aggttacgat tggttgatta tgacacccgg tgtgggttta 2761 gatgacaagg gagatgcatt gggtcaacag tatagaaccg tggatgatgt ggtttctaca 2821 ggatctgaca ttattattgt tggaagagga ctatttgcaa agggaaggga tgctaaggta 2881 gagggtgaac gttacagaaa agcaggctgg gaagcatatt tgagaagatg cggccagcaa 2941 aactaaaaaa ctgtattata agtaaatgca tgtatactaa actcacaaat tagagcttca 3001 atttaattat atcagttatt acccgagtaa cgaagcatct gtgcttcatt ttgtagaaca 3061 aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga 3121 acagaaatgc aacgcgaaag cgctatttta ccaacgaaga atctgtgctt catttttgta 3181 aaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt 3241 acagaacaga aatgcaacgc gagagcgcta ttttaccaac aaagaatcta tacttctttt 3301 ttgttctaca aaaatgcatc ccgagagcgc tatttttcta acaaagcatc ttagattact 3361 ttttttctcc tttgtgcgct ctataatgca gtctcttgat aactttttgc actgtaggtc 3421 cgttaaggtt agaagaaggc tactttggtg tctattttct cttccataaa aaaagcctga 3481 ctccacttcc cgcgtttact gattactagc gaagctgcgg gtgcattttt tcaagataaa 3541 ggcatccccg attatattct ataccgatgt ggattgcgca tactttgtga acagaaagtg 3601 atagcgttga tgattcttca ttggtcagaa aattatgaac ggtttcttct attttgtctc 3661 tatatactac gtataggaaa tgtttacatt ttcgtattgt tttcgattca ctctatgaat 3721 agttcttact acaatttttt tgtctaaaga gtaatactag agataaacat aaaaaatgta 3781 gaggtcgagt ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt tatataggga 3841 tatagcacag agatatatag caaagagata cttttgagca atgtttgtgg aagcggtatt 3901 cgcaatattt tagtagctcg ttacagtccg gtgcgttttt ggttttttga aagtgcgtca 3961 tcagagcgct tttggttttc aaaagcgctc tgaagttcct atactttcta gctagagaat 4021 aggaacttcc cgagcggccg cgtgttacaa gcttgactcc agcgtaactg gactgcaatc 4081 aactcactgg ctcaccttca cgggtgggcc tttcttcggt agaaaatcaa aggatcttct 4141 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca 4201 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaggta actggcttca 4261 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca 4321 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg 4381 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg 4441 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct 4501 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga 4561 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc 4621 ttccaggggt aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg 4681 agcatcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg 4741 cagaaaggcc cacccgaagg tgagccaggt gattacattt gggccctcat cagaggtttt 4801 caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa 4861 gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg 4921 ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca 4981 cttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca 5041 tagttgcctg actccccgtc gtgtagataa ctacgatgcg ggagggctta ccatctggcc 5101 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa 5161 accagccagc cgggagggcc gagcgcagaa gtgatcctgc aactttatcc gcctccatcc 5221 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca 5281 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat 5341 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag 5401 cggttagctc cttcggtcct ccgatcgttg ccagaagtaa gttggccgca gtgttatcac 5461 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgtg agatgctttt 5521 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt 5581 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc 5641 tcatcattgg aaaacgttct tcggggcgta aactctcaag gatcttaccg ctgttgagat 5701 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca 5761 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga 5821 cacggaaatg ttgaatactc attttagctt ccttagctcc tgaaaatctc gataactcaa 5881 aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc 5941 gatcaagtca aaagcctccg gtcggaggct tttgactttc tgctatggag gtcaggtatg 6001 atttaaatgg tcagttaga //