LOCUS Exported 7519 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Yeast expression plasmid for SARS-CoV-2 RNA-dependent RNA polymerase. ACCESSION . VERSION . KEYWORDS pGBW-m4046618 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7519) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 7519) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7519 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(2857..2875) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 3226..4029 /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 4112..5094 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 5172..5201 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 5223..5810 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 5822..5849 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(5873..6064) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(6065..6925) /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 6688..6707 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6926..7028) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 7029..7060 /label=tonB terminator /note="bidirectional E. coli tonB-P14 transcription terminator" promoter 7112..7519 /gene="S. cerevisiae TEF1" /label=TEF1 promoter /note="promoter for EF-1-alpha" ORIGIN 1 ttttaattac aaaatgtccg ccgatgcaca atctttctta aatagagtct gcggcgtgag 61 tgctgctcgt ctaacccctt gtggtacggg tacttcaaca gacgtagttt acagggcgtt 121 tgatatttat aacgacaagg tcgctggttt cgctaaattt ttgaagacca actgttgtag 181 attccaggag aaggatgaag acgacaatct tatcgattcg tacttcgttg ttaaaagaca 241 tactttttcc aactatcaac acgaagaaac aatatacaat ttgctcaagg actgtccagc 301 cgtcgctaaa cacgattttt tcaagttcag aattgatggg gacatggtgc cacatatcag 361 cagacaacgg ttgaccaagt acactatggc tgatttagtt tacgccttgc gacactttga 421 cgaaggaaac tgtgatactc tgaaggaaat tttggttacc tataactgct gtgacgatga 481 ttacttcaac aaaaaggact ggtacgactt cgtagagaat cccgacatct tgagggttta 541 cgctaactta ggtgaaagag tcagacaagc tttgttaaag acagtccaat tttgcgatgc 601 tatgagaaat gcgggtattg ttggtgtttt gactttggat aatcaagatt tgaacggtaa 661 ctggtatgac ttcggtgatt ttatccagac taccccgggt tctggggtac cagttgtcga 721 ttcttactat tctctgttaa tgccaatact caccctaact agagcattga ctgccgaatc 781 ccatgttgac actgatttaa ctaagccata cattaaatgg gaccttctaa agtacgattt 841 caccgaagaa cgtttaaagt tgttcgacag atattttaaa tactgggacc aaacctacca 901 ccctaattgt gtcaactgtt tggacgacag atgtattctt cactgcgcta actttaatgt 961 gttgttctcg actgtttttc caccaacctc tttcggtccg ctggttcgaa agatttttgt 1021 tgacggtgtt ccattcgtgg tcagtacggg atatcatttc agagaattag gggtcgttca 1081 taaccaagac gtgaacttgc actcatcaag attgtccttc aaggaattgt tggtctacgc 1141 agctgatcct gcaatgcacg ccgcttctgg aaacttattg ctggataaga gaacaacctg 1201 tttcagcgtt gccgctctta ctaataacgt cgcgttccaa actgtaaagc cgggtaactt 1261 taataaagac ttctacgatt ttgctgttag caagggtttt ttcaaagaag gttctagtgt 1321 tgagttgaaa cactttttct ttgctcaaga cggcaatgct gctatcagtg attatgacta 1381 ttaccgttac aacttaccaa caatgtgtga tatcagacag ttgcttttcg tcgtcgaagt 1441 tgttgataaa tattttgact gttatgatgg tggttgcatc aatgcaaatc aagtcattgt 1501 caacaactta gataagtctg ctggcttccc attcaacaag tggggtaaag ccagattgta 1561 ctatgattcc atgtcctacg aggaccaaga tgctttgttc gcgtacacca agagaaacgt 1621 tattcctaca attactcaaa tgaatctaaa gtacgctatc tctgcaaaaa atcgggccag 1681 aaccgttgct ggtgtttcta tctgttctac gatgactaac cgtcaatttc atcagaagct 1741 attgaaatcc atagctgcta ctagaggtgc tactgtagtc atcggtacct caaagttcta 1801 tggtggttgg cacaatatgc tcaagaccgt ttactctgat gtcgaaaacc cacatcttat 1861 gggttgggac taccctaagt gtgatagagc catgccaaat atgctaagaa ttatggcatc 1921 cttggtcttg gctagaaagc ataccacatg ctgctcattg tcccaccgtt tttacagatt 1981 ggccaacgaa tgtgctcaag tcctttctga aatggttatg tgtggtggta gtctatatgt 2041 aaagccaggc ggaacttcgt ctggagatgc caccactgct tacgctaact cggtttttaa 2101 catttgtcaa gccgttaccg ccaacgtaaa cgctttgctg tctacggatg gtaacaagat 2161 cgctgataag tatgtgcgca acttgcaaca tagactttat gaatgtttgt atagaaatcg 2221 agatgtcgat actgacttcg ttaacgaatt ttacgcgtac ttaagaaagc acttctccat 2281 gatgattttg tcagacgacg ctgttgtttg tttcaattct acttacgcta gccaaggttt 2341 ggtcgcttca atcaaaaact tcaagtctgt tttatactac caaaacaatg tttttatgtc 2401 cgaggcaaaa tgctggacgg aaacagattt gactaagggt ccacatgaat tttgtagtca 2461 acacacaatg ttagtgaaac aaggggacga ctacgtttac ctaccctacc cagatccaag 2521 cagaattttg ggtgccggtt gtttcgtgga cgacatagtc aagaccgacg gtaccttgat 2581 gatcgaacgg tttgtctctc tggccataga cgcttaccca ttgactaagc accctaacca 2641 agaatatgct gatgtttttc atttatatct tcaatacatt agaaaattac atgacgaatt 2701 gaccggtcac atgttagata tgtactctgt gatgttgacg aacgataata cgtcacgtta 2761 ctgggaaccc gaattttacg aagcgatgta cactccacac acagtattgc agtaaacagg 2821 ccccttttcc tttgtcgata tcatgtaatt agttatgtca cgcttacatt cacgccctcc 2881 tcccacatcc gctctaaccg aaaaggaagg agttagacaa cctgaagtct aggtccctat 2941 ttattttttt taatagttat gttagtatta agaacgttat ttatatttca aatttttctt 3001 ttttttctgt acaaagcggc cgccgaacca ctagtgcact gcagtacacg gtttccttga 3061 aatttttttg attcggtaat ctccgaacag aaggaagaac gaaggaagga gcacagactt 3121 agattggtat atatacgcat atgtagtgtt gaagaaacat gaaattgccc agtattctta 3181 acccaactgc acagaacaaa aacgtgcagg aaacgaagat aaatcatgtc gaaagctaca 3241 tataaggaac gtgctgctac tcatcctagt cctgttgctg ccaagctatt taatatcatg 3301 cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc gtaccaccaa ggaattactg 3361 gagttagttg aagcattagg tcccaaaatt tgtttactaa aaacacatgt ggatatcttg 3421 actgattttt ccatggaggg cacagttaag ccgctaaagg cattatccgc caagtacaat 3481 tttttactct tcgaggacag aaaatttgct gacattggta atacagtcaa attgcagtac 3541 tctgcgggtg tatacagaat agcagaatgg gcagacatta cgaatgcaca cggtgtggtg 3601 ggcccaggta ttgttagcgg tttgaagcag gcggcagaag aagtaacaaa ggaacctaga 3661 ggccttttga tgttagcaga attgtcatgc aagggctccc tatctactgg agaatatact 3721 aagggtactg ttgacattgc gaagagcgac aaagattttg ttatcggctt tattgctcaa 3781 agagacatgg gtggaagaga tgaaggttac gattggttga ttatgacacc cggtgtgggt 3841 ttagatgaca agggagatgc attgggtcaa cagtatagaa ccgtggatga tgtggtttct 3901 acaggatctg acattattat tgttggaaga ggactatttg caaagggaag ggatgctaag 3961 gtagagggtg aacgttacag aaaagcaggc tgggaagcat atttgagaag atgcggccag 4021 caaaactaaa aaactgtatt ataagtaaat gcatgtatac taaactcaca aattagagct 4081 tcaatttaat tatatcagtt attacccgag taacgaagca tctgtgcttc attttgtaga 4141 acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac 4201 agaacagaaa tgcaacgcga aagcgctatt ttaccaacga agaatctgtg cttcattttt 4261 gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt 4321 tttacagaac agaaatgcaa cgcgagagcg ctattttacc aacaaagaat ctatacttct 4381 tttttgttct acaaaaatgc atcccgagag cgctattttt ctaacaaagc atcttagatt 4441 actttttttc tcctttgtgc gctctataat gcagtctctt gataactttt tgcactgtag 4501 gtccgttaag gttagaagaa ggctactttg gtgtctattt tctcttccat aaaaaaagcc 4561 tgactccact tcccgcgttt actgattact agcgaagctg cgggtgcatt ttttcaagat 4621 aaaggcatcc ccgattatat tctataccga tgtggattgc gcatactttg tgaacagaaa 4681 gtgatagcgt tgatgattct tcattggtca gaaaattatg aacggtttct tctattttgt 4741 ctctatatac tacgtatagg aaatgtttac attttcgtat tgttttcgat tcactctatg 4801 aatagttctt actacaattt ttttgtctaa agagtaatac tagagataaa cataaaaaat 4861 gtagaggtcg agtttagatg caagttcaag gagcgaaagg tggatgggta ggttatatag 4921 ggatatagca cagagatata tagcaaagag atacttttga gcaatgtttg tggaagcggt 4981 attcgcaata ttttagtagc tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg 5041 tcatcagagc gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt ctagctagag 5101 aataggaact tcccgagcgg ccgcgtgtta caagcttgac tccagcgtaa ctggactgca 5161 atcaactcac tggctcacct tcacgggtgg gcctttcttc ggtagaaaat caaaggatct 5221 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta 5281 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgag gtaactggct 5341 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact 5401 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg 5461 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata 5521 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga 5581 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag 5641 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg 5701 agcttccagg ggtaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac 5761 ttgagcatcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca 5821 acgcagaaag gcccacccga aggtgagcca ggtgattaca tttgggccct catcagaggt 5881 tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt 5941 gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa 6001 gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg 6061 tcacttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat 6121 ccatagttgc ctgactcccc gtcgtgtaga taactacgat gcgggagggc ttaccatctg 6181 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa 6241 taaaccagcc agccgggagg gccgagcgca gaagtgatcc tgcaacttta tccgcctcca 6301 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc 6361 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt 6421 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa 6481 aagcggttag ctccttcggt cctccgatcg ttgccagaag taagttggcc gcagtgttat 6541 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtgagatgct 6601 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga 6661 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag 6721 tgctcatcat tggaaaacgt tcttcggggc gtaaactctc aaggatctta ccgctgttga 6781 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca 6841 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg 6901 cgacacggaa atgttgaata ctcattttag cttccttagc tcctgaaaat ctcgataact 6961 caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt 7021 gccgatcaag tcaaaagcct ccggtcggag gcttttgact ttctgctatg gaggtcaggt 7081 atgatttaaa tggtcagtta gagcggccgc cccacacacc atagcttcaa aatgtttcta 7141 ctcctttttt actcttccag attttctcgg actccgcgca tcgccgtacc acttcaaaac 7201 acccaagcac agcatactaa atttcccctc tttcttcctc tagggtgtcg ttaattaccc 7261 gtactaaagg tttggaaaag aaaaaagaga ccgcctcgtt tctttttctt cgtcgaaaaa 7321 ggcaataaaa atttttatca cgtttctttt tcttgaaaat tttttttttt gatttttttc 7381 tctttcgatg acctcccatt gatatttaag ttaataaacg gtcttcaatt tctcaagttt 7441 cagtttcatt tttcttgttc tattacaact ttttttactt cttgctcatt agaaagaaag 7501 catagcaatc taatctaag //