LOCUS Exported 6013 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Yeast Expression plasmid for SARS-CoV-2 N (nucleocapsid). ACCESSION . VERSION . KEYWORDS pGBW-m4046174 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6013) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 6013) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6013 /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 1688..1705 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" CDS 1706..1723 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(1768..1786) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 2137..2940 /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 3023..4005 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 4083..4112 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 4134..4721 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 4733..4760 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(4784..4975) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(4976..5836) /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 5599..5618 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5837..5939) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 5940..5971 /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 atggacccca aaatcagcga aatgcacccc gcattacgtt 481 tggtggaccc tcagattcaa ctggcagtaa ccagaatgga gaacgcagtg gggcgcgatc 541 aaaacaacgt cggccccaag gtttacccaa taatactgcg tcttggttca ccgctctcac 601 tcaacatggc aaggaagacc ttaaattccc tcgaggacaa ggcgttccaa ttaacaccaa 661 tagcagtcca gatgaccaaa ttggctacta ccgaagagct accagacgaa ttcgtggtgg 721 tgacggtaaa atgaaagatc tcagtccaag atggtatttc tactacctag gaactgggcc 781 agaagctgga cttccctatg gtgctaacaa agacggcatc atatgggttg caactgaggg 841 agccttgaat acaccaaaag atcacattgg cacccgcaat cctgctaaca atgctgcaat 901 cgtgctacaa cttcctcaag gaacaacatt gccaaaaggc ttctacgcag aagggagcag 961 aggcggcagt caagcctctt ctcgttcctc atcacgtagt cgcaacagtt caagaaattc 1021 aactccaggc agcagtaggg gaacttctcc tgctagaatg gctggcaatg gcggtgatgc 1081 tgctcttgct ttgctgctgc ttgacagatt gaaccagctt gagagcaaaa tgtctggtaa 1141 aggccaacaa caacaaggcc aaactgtcac taagaaatct gctgctgagg cttctaagaa 1201 gcctcggcaa aaacgtactg ccactaaagc atacaatgta acacaagctt tcggcagacg 1261 tggtccagaa caaacccaag gaaattttgg ggaccaggaa ctaatcagac aaggaactga 1321 ttacaaacat tggccgcaaa ttgcacaatt tgcccccagc gcttcagcgt tcttcggaat 1381 gtcgcgcatt ggcatggaag tcacaccttc gggaacgtgg ttgacctaca caggtgccat 1441 caaattggat gacaaagatc caaatttcaa agatcaagtc attttgctga ataagcatat 1501 tgacgcatac aaaacattcc caccaacaga gcctaaaaag gacaaaaaga agaaggctga 1561 tgaaactcaa gccttaccgc agagacagaa gaaacagcaa actgtgactc ttcttcctgc 1621 tgcagatttg gatgatttct ccaaacaatt gcaacaatcc atgagcagtg ctgactcaac 1681 tcaggccctc gtgccgagag gttctcacca tcatcaccat cattaaacag gccccttttc 1741 ctttgtcgat atcatgtaat tagttatgtc acgcttacat tcacgccctc ctcccacatc 1801 cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt 1861 ttaatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg 1921 tacaaagcgg ccgccgaacc actagtgcac tgcagtacac ggtttccttg aaattttttt 1981 gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg agcacagact tagattggta 2041 tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt aacccaactg 2101 cacagaacaa aaacgtgcag gaaacgaaga taaatcatgt cgaaagctac atataaggaa 2161 cgtgctgcta ctcatcctag tcctgttgct gccaagctat ttaatatcat gcacgaaaag 2221 caaacaaact tgtgtgcttc attggatgtt cgtaccacca aggaattact ggagttagtt 2281 gaagcattag gtcccaaaat ttgtttacta aaaacacatg tggatatctt gactgatttt 2341 tccatggagg gcacagttaa gccgctaaag gcattatccg ccaagtacaa ttttttactc 2401 ttcgaggaca gaaaatttgc tgacattggt aatacagtca aattgcagta ctctgcgggt 2461 gtatacagaa tagcagaatg ggcagacatt acgaatgcac acggtgtggt gggcccaggt 2521 attgttagcg gtttgaagca ggcggcagaa gaagtaacaa aggaacctag aggccttttg 2581 atgttagcag aattgtcatg caagggctcc ctatctactg gagaatatac taagggtact 2641 gttgacattg cgaagagcga caaagatttt gttatcggct ttattgctca aagagacatg 2701 ggtggaagag atgaaggtta cgattggttg attatgacac ccggtgtggg tttagatgac 2761 aagggagatg cattgggtca acagtataga accgtggatg atgtggtttc tacaggatct 2821 gacattatta ttgttggaag aggactattt gcaaagggaa gggatgctaa ggtagagggt 2881 gaacgttaca gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa 2941 aaaactgtat tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa 3001 ttatatcagt tattacccga gtaacgaagc atctgtgctt cattttgtag aacaaaaatg 3061 caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa 3121 atgcaacgcg aaagcgctat tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa 3181 aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa 3241 cagaaatgca acgcgagagc gctattttac caacaaagaa tctatacttc ttttttgttc 3301 tacaaaaatg catcccgaga gcgctatttt tctaacaaag catcttagat tacttttttt 3361 ctcctttgtg cgctctataa tgcagtctct tgataacttt ttgcactgta ggtccgttaa 3421 ggttagaaga aggctacttt ggtgtctatt ttctcttcca taaaaaaagc ctgactccac 3481 ttcccgcgtt tactgattac tagcgaagct gcgggtgcat tttttcaaga taaaggcatc 3541 cccgattata ttctataccg atgtggattg cgcatacttt gtgaacagaa agtgatagcg 3601 ttgatgattc ttcattggtc agaaaattat gaacggtttc ttctattttg tctctatata 3661 ctacgtatag gaaatgttta cattttcgta ttgttttcga ttcactctat gaatagttct 3721 tactacaatt tttttgtcta aagagtaata ctagagataa acataaaaaa tgtagaggtc 3781 gagtttagat gcaagttcaa ggagcgaaag gtggatgggt aggttatata gggatatagc 3841 acagagatat atagcaaaga gatacttttg agcaatgttt gtggaagcgg tattcgcaat 3901 attttagtag ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc gtcatcagag 3961 cgcttttggt tttcaaaagc gctctgaagt tcctatactt tctagctaga gaataggaac 4021 ttcccgagcg gccgcgtgtt acaagcttga ctccagcgta actggactgc aatcaactca 4081 ctggctcacc ttcacgggtg ggcctttctt cggtagaaaa tcaaaggatc ttcttgagat 4141 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg 4201 gtttgtttgc cggatcaaga gctaccaact ctttttccga ggtaactggc ttcagcagag 4261 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact 4321 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg 4381 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc 4441 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg 4501 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg 4561 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag 4621 gggtaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcatc 4681 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcagaaa 4741 ggcccacccg aaggtgagcc aggtgattac atttgggccc tcatcagagg ttttcaccgt 4801 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt 4861 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg 4921 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcacttacc 4981 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg 5041 cctgactccc cgtcgtgtag ataactacga tgcgggaggg cttaccatct ggccccagtg 5101 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc 5161 cagccgggag ggccgagcgc agaagtgatc ctgcaacttt atccgcctcc atccagtcta 5221 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg 5281 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct 5341 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta 5401 gctccttcgg tcctccgatc gttgccagaa gtaagttggc cgcagtgtta tcactcatgg 5461 ttatggcagc actgcataat tctcttactg tcatgccatc cgtgagatgc ttttctgtga 5521 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt 5581 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca 5641 ttggaaaacg ttcttcgggg cgtaaactct caaggatctt accgctgttg agatccagtt 5701 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt 5761 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga 5821 aatgttgaat actcatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata 5881 cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa 5941 gtcaaaagcc tccggtcgga ggcttttgac tttctgctat ggaggtcagg tatgatttaa 6001 atggtcagtt aga //