LOCUS Exported 7093 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Yeast Expression plasmid for SARS-CoV-2 N (nucleocapsid). ACCESSION . VERSION . KEYWORDS pGBW-m4046575 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7093) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 7093) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7093 /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" primer_bind complement(2848..2866) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 3217..4020 /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 4103..5085 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 5163..5192 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 5214..5801 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 5813..5840 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(5864..6055) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(6056..6916) /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 6679..6698 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" gap 6935..6944 /estimated_length=10 terminator 7020..7051 /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 gttctaaaat cgaggaaggt aagctagtca tttggataaa 1741 tggagataag ggttacaacg gtttggcaga agtaggcaag aaatttgaaa aggacacagg 1801 tatcaaggtg acggttgaac atccagacaa gttagaagaa aaattccctc aagttgctgc 1861 tactggtgat ggtcccgaca ttattttctg ggcccacgat cgttttggtg gatatgctca 1921 gtcaggtttg ctggctgaga tcaccccaga caaggccttc caagataaat tgtacccatt 1981 cacttgggat gctgtcagat acaacggtaa gcttatagca tatccaattg ctgttgaagc 2041 cttatcgttg atctacaata aggacttgtt acctaaccca ccaaagacat gggaagaaat 2101 tccagctttg gataaagagc taaaggccaa gggaaaaagc gctttgatgt ttaacttaca 2161 agaaccttat ttcacctggc cattaatcgc tgctgacggt ggttacgcat ttaaatacga 2221 aaacggtaag tatgacatta aggatgtcgg cgttgataat gctggtgcaa aggctggttt 2281 gaccttcttg gttgatttaa taaagaataa acacatgaac gctgacactg actactctat 2341 cgccgaagcc gctttcaaca agggtgaaac tgctatgacc ataaatggtc catgggcctg 2401 gagtaacatt gatacttcta aagtaaacta tggtgtcact gttttgccta cattcaaagg 2461 ccaaccatcc aagccctttg tcggtgttct atccgctggc atcaatgctg cttctccaaa 2521 caaggagcta gctaaagaat ttttggaaaa ttacttgtta actgacgaag gtttggaggc 2581 cgttaacaag gacaagccat tgggtgctgt cgctttgaaa tcttacgaag aagaattggc 2641 taaggatccc agaattgccg ctaccatgga gaacgcccaa aagggtgaaa ttatgccaaa 2701 catcccacaa atgtcagcct tctggtacgc agttaggact gccgtgatta atgctgcttc 2761 cggtagacaa accgtcgacg aagcattaaa agatgctcaa acgtaaacag gccccttttc 2821 ctttgtcgat atcatgtaat tagttatgtc acgcttacat tcacgccctc ctcccacatc 2881 cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt 2941 ttaatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg 3001 tacaaagcgg ccgccgaacc actagtgcac tgcagtacac ggtttccttg aaattttttt 3061 gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg agcacagact tagattggta 3121 tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt aacccaactg 3181 cacagaacaa aaacgtgcag gaaacgaaga taaatcatgt cgaaagctac atataaggaa 3241 cgtgctgcta ctcatcctag tcctgttgct gccaagctat ttaatatcat gcacgaaaag 3301 caaacaaact tgtgtgcttc attggatgtt cgtaccacca aggaattact ggagttagtt 3361 gaagcattag gtcccaaaat ttgtttacta aaaacacatg tggatatctt gactgatttt 3421 tccatggagg gcacagttaa gccgctaaag gcattatccg ccaagtacaa ttttttactc 3481 ttcgaggaca gaaaatttgc tgacattggt aatacagtca aattgcagta ctctgcgggt 3541 gtatacagaa tagcagaatg ggcagacatt acgaatgcac acggtgtggt gggcccaggt 3601 attgttagcg gtttgaagca ggcggcagaa gaagtaacaa aggaacctag aggccttttg 3661 atgttagcag aattgtcatg caagggctcc ctatctactg gagaatatac taagggtact 3721 gttgacattg cgaagagcga caaagatttt gttatcggct ttattgctca aagagacatg 3781 ggtggaagag atgaaggtta cgattggttg attatgacac ccggtgtggg tttagatgac 3841 aagggagatg cattgggtca acagtataga accgtggatg atgtggtttc tacaggatct 3901 gacattatta ttgttggaag aggactattt gcaaagggaa gggatgctaa ggtagagggt 3961 gaacgttaca gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa 4021 aaaactgtat tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa 4081 ttatatcagt tattacccga gtaacgaagc atctgtgctt cattttgtag aacaaaaatg 4141 caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa 4201 atgcaacgcg aaagcgctat tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa 4261 aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa 4321 cagaaatgca acgcgagagc gctattttac caacaaagaa tctatacttc ttttttgttc 4381 tacaaaaatg catcccgaga gcgctatttt tctaacaaag catcttagat tacttttttt 4441 ctcctttgtg cgctctataa tgcagtctct tgataacttt ttgcactgta ggtccgttaa 4501 ggttagaaga aggctacttt ggtgtctatt ttctcttcca taaaaaaagc ctgactccac 4561 ttcccgcgtt tactgattac tagcgaagct gcgggtgcat tttttcaaga taaaggcatc 4621 cccgattata ttctataccg atgtggattg cgcatacttt gtgaacagaa agtgatagcg 4681 ttgatgattc ttcattggtc agaaaattat gaacggtttc ttctattttg tctctatata 4741 ctacgtatag gaaatgttta cattttcgta ttgttttcga ttcactctat gaatagttct 4801 tactacaatt tttttgtcta aagagtaata ctagagataa acataaaaaa tgtagaggtc 4861 gagtttagat gcaagttcaa ggagcgaaag gtggatgggt aggttatata gggatatagc 4921 acagagatat atagcaaaga gatacttttg agcaatgttt gtggaagcgg tattcgcaat 4981 attttagtag ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc gtcatcagag 5041 cgcttttggt tttcaaaagc gctctgaagt tcctatactt tctagctaga gaataggaac 5101 ttcccgagcg gccgcgtgtt acaagcttga ctccagcgta actggactgc aatcaactca 5161 ctggctcacc ttcacgggtg ggcctttctt cggtagaaaa tcaaaggatc ttcttgagat 5221 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg 5281 gtttgtttgc cggatcaaga gctaccaact ctttttccga ggtaactggc ttcagcagag 5341 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact 5401 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg 5461 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc 5521 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg 5581 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg 5641 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag 5701 gggtaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcatc 5761 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcagaaa 5821 ggcccacccg aaggtgagcc aggtgattac atttgggccc tcatcagagg ttttcaccgt 5881 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt 5941 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg 6001 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcacttacc 6061 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg 6121 cctgactccc cgtcgtgtag ataactacga tgcgggaggg cttaccatct ggccccagtg 6181 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc 6241 cagccgggag ggccgagcgc agaagtgatc ctgcaacttt atccgcctcc atccagtcta 6301 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg 6361 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct 6421 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta 6481 gctccttcgg tcctccgatc gttgccagaa gtaagttggc cgcagtgtta tcactcatgg 6541 ttatggcagc actgcataat tctcttactg tcatgccatc cgtgagatgc ttttctgtga 6601 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt 6661 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca 6721 ttggaaaacg ttcttcgggg cgtaaactct caaggatctt accgctgttg agatccagtt 6781 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt 6841 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga 6901 aatgttgaat actcatttta gcttccttag ctccnnnnnn nnnngataac tcaaaaaata 6961 cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa 7021 gtcaaaagcc tccggtcgga ggcttttgac tttctgctat ggaggtcagg tatgatttaa 7081 atggtcagtt aga //