LOCUS Exported 8676 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Yeast Expression plasmid for SARS-CoV-2 RNA-dependent RNA polymerase. ACCESSION . VERSION . KEYWORDS pGBW-m4046514 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8676) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 8676) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8676 /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 3271..3288 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" primer_bind complement(4431..4449) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 4800..5603 /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 5686..6668 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 6746..6775 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 6797..7384 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 7396..7423 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(7447..7638) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(7639..8499) /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 8262..8281 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8500..8602) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 8603..8634 /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 gctgatgcac aatcgttttt aaaccgggtt tgcggtgtaa gtgcagcccg tcttacaccg 541 tgcggcacag gcactagtac tgatgtcgta tacagggctt ttgacatcta caatgataaa 601 gtagctggtt ttgctaaatt cctaaaaact aattgttgtc gcttccaaga aaaggacgaa 661 gatgacaatt taattgattc ttactttgta gttaagagac acactttctc taactaccaa 721 catgaagaaa caatttataa tttacttaag gattgtccag ctgttgctaa acatgacttc 781 tttaagttta gaatagacgg tgacatggta ccacatatat cacgtcaacg tcttactaaa 841 tacacaatgg cagacctcgt ctatgcttta aggcattttg atgaaggtaa ttgtgacaca 901 ttaaaagaaa tacttgtcac atacaattgt tgtgatgatg attatttcaa taaaaaggac 961 tggtatgatt ttgtagaaaa cccagatata ttacgcgtat acgccaactt aggtgaacgt 1021 gtacgccaag ctttgttaaa aacagtacaa ttctgtgatg ccatgcgaaa tgctggtatt 1081 gttggtgtac tgacattaga taatcaagat ctcaatggta actggtatga tttcggtgat 1141 ttcatacaaa ccacgccagg tagtggagtt cctgttgtag attcttatta ttcattgtta 1201 atgcctatat taaccttgac cagggcttta actgcagagt cacatgttga cactgactta 1261 acaaagcctt acattaagtg ggatttgtta aaatatgact tcacggaaga gaggttaaaa 1321 ctctttgacc gttattttaa atattgggat cagacatacc acccaaattg tgttaactgt 1381 ttggatgaca gatgcattct gcattgtgca aactttaatg ttttattctc tacagtgttc 1441 ccacctacaa gttttggacc actagtgaga aaaatatttg ttgatggtgt tccatttgta 1501 gtttcaactg gataccactt cagagagcta ggtgttgtac ataatcagga tgtaaactta 1561 catagctcta gacttagttt taaggaatta cttgtgtatg ctgctgaccc tgctatgcac 1621 gctgcttctg gtaatctatt actagataaa cgcactacgt gcttttcagt agctgcactt 1681 actaacaatg ttgcttttca aactgtcaaa cccggtaatt ttaacaaaga cttctatgac 1741 tttgctgtgt ctaagggttt ctttaaggaa ggaagttctg ttgaattaaa acacttcttc 1801 tttgctcagg atggtaatgc tgctatcagc gattatgact actatcgtta taatctacca 1861 acaatgtgtg atatcagaca actactattt gtagttgaag ttgttgataa gtactttgat 1921 tgttacgatg gtggctgtat taatgctaac caagtcatcg tcaacaacct agacaaatca 1981 gctggttttc catttaataa atggggtaag gctagacttt attatgattc aatgagttat 2041 gaggatcaag atgcactttt cgcatataca aaacgtaatg tcatccctac tataactcaa 2101 atgaatctta agtatgccat tagtgcaaag aatagagctc gcaccgtagc tggtgtctct 2161 atctgtagta ctatgaccaa tagacagttt catcaaaaat tattgaaatc aatagccgcc 2221 actagaggag ctactgtagt aattggaaca agcaaattct atggtggttg gcacaacatg 2281 ttaaaaactg tttatagtga tgtagaaaac cctcacctta tgggttggga ttatcctaaa 2341 tgtgatagag ccatgcctaa catgcttaga attatggcct cacttgttct tgctcgcaaa 2401 catacaacgt gttgtagctt gtcacaccgt ttctatagat tagctaatga gtgtgctcaa 2461 gtattgagtg aaatggtcat gtgtggcggt tcactatatg ttaaaccagg tggaacctca 2521 tcaggagatg ccacaactgc ttatgctaat agtgttttta acatttgtca agctgtcacg 2581 gccaatgtta atgcactttt atctactgat ggtaacaaaa ttgccgataa gtatgtccgc 2641 aatttacaac acagacttta tgagtgtctc tatagaaata gagatgttga cacagacttt 2701 gtgaatgagt tttacgcata tttgcgtaaa catttctcaa tgatgatact ctctgacgat 2761 gctgttgtgt gtttcaatag cacttatgca tctcaaggtc tagtggctag cataaagaac 2821 tttaagtcag ttctttatta tcaaaacaat gtttttatgt ctgaagcaaa atgttggact 2881 gagactgacc ttactaaagg acctcatgaa ttttgctctc aacatacaat gctagttaaa 2941 cagggtgatg attatgtgta ccttccttac ccagatccat caagaatcct aggggccggc 3001 tgttttgtag atgatatcgt aaaaacagat ggtacactta tgattgaacg gttcgtgtct 3061 ttagctatag atgcttaccc acttactaaa catcctaatc aggagtatgc tgatgtcttt 3121 catttgtact tacaatacat aagaaagcta catgatgagt taacaggaca catgttagac 3181 atgtattctg ttatgcttac taatgataac acttcaaggt attgggaacc tgagttttat 3241 gaggctatgt acacaccgca tacagtctta ctcgtgccga gaggttctaa aatcgaggaa 3301 ggtaagctag tcatttggat aaatggagat aagggttaca acggtttggc agaagtaggc 3361 aagaaatttg aaaaggacac aggtatcaag gtgacggttg aacatccaga caagttagaa 3421 gaaaaattcc ctcaagttgc tgctactggt gatggtcccg acattatttt ctgggcccac 3481 gatcgttttg gtggatatgc tcagtcaggt ttgctggctg agatcacccc agacaaggcc 3541 ttccaagata aattgtaccc attcacttgg gatgctgtca gatacaacgg taagcttata 3601 gcatatccaa ttgctgttga agccttatcg ttgatctaca ataaggactt gttacctaac 3661 ccaccaaaga catgggaaga aattccagct ttggataaag agctaaaggc caagggaaaa 3721 agcgctttga tgtttaactt acaagaacct tatttcacct ggccattaat cgctgctgac 3781 ggtggttacg catttaaata cgaaaacggt aagtatgaca ttaaggatgt cggcgttgat 3841 aatgctggtg caaaggctgg tttgaccttc ttggttgatt taataaagaa taaacacatg 3901 aacgctgaca ctgactactc tatcgccgaa gccgctttca acaagggtga aactgctatg 3961 accataaatg gtccatgggc ctggagtaac attgatactt ctaaagtaaa ctatggtgtc 4021 actgttttgc ctacattcaa aggccaacca tccaagccct ttgtcggtgt tctatccgct 4081 ggcatcaatg ctgcttctcc aaacaaggag ctagctaaag aatttttgga aaattacttg 4141 ttaactgacg aaggtttgga ggccgttaac aaggacaagc cattgggtgc tgtcgctttg 4201 aaatcttacg aagaagaatt ggctaaggat cccagaattg ccgctaccat ggagaacgcc 4261 caaaagggtg aaattatgcc aaacatccca caaatgtcag ccttctggta cgcagttagg 4321 actgccgtga ttaatgctgc ttccggtaga caaaccgtcg acgaagcatt aaaagatgct 4381 caaacgtaaa caggcccctt ttcctttgtc gatatcatgt aattagttat gtcacgctta 4441 cattcacgcc ctcctcccac atccgctcta accgaaaagg aaggagttag acaacctgaa 4501 gtctaggtcc ctatttattt tttttaatag ttatgttagt attaagaacg ttatttatat 4561 ttcaaatttt tctttttttt ctgtacaaag cggccgccga accactagtg cactgcagta 4621 cacggtttcc ttgaaatttt tttgattcgg taatctccga acagaaggaa gaacgaagga 4681 aggagcacag acttagattg gtatatatac gcatatgtag tgttgaagaa acatgaaatt 4741 gcccagtatt cttaacccaa ctgcacagaa caaaaacgtg caggaaacga agataaatca 4801 tgtcgaaagc tacatataag gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc 4861 tatttaatat catgcacgaa aagcaaacaa acttgtgtgc ttcattggat gttcgtacca 4921 ccaaggaatt actggagtta gttgaagcat taggtcccaa aatttgttta ctaaaaacac 4981 atgtggatat cttgactgat ttttccatgg agggcacagt taagccgcta aaggcattat 5041 ccgccaagta caatttttta ctcttcgagg acagaaaatt tgctgacatt ggtaatacag 5101 tcaaattgca gtactctgcg ggtgtataca gaatagcaga atgggcagac attacgaatg 5161 cacacggtgt ggtgggccca ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa 5221 caaaggaacc tagaggcctt ttgatgttag cagaattgtc atgcaagggc tccctatcta 5281 ctggagaata tactaagggt actgttgaca ttgcgaagag cgacaaagat tttgttatcg 5341 gctttattgc tcaaagagac atgggtggaa gagatgaagg ttacgattgg ttgattatga 5401 cacccggtgt gggtttagat gacaagggag atgcattggg tcaacagtat agaaccgtgg 5461 atgatgtggt ttctacagga tctgacatta ttattgttgg aagaggacta tttgcaaagg 5521 gaagggatgc taaggtagag ggtgaacgtt acagaaaagc aggctgggaa gcatatttga 5581 gaagatgcgg ccagcaaaac taaaaaactg tattataagt aaatgcatgt atactaaact 5641 cacaaattag agcttcaatt taattatatc agttattacc cgagtaacga agcatctgtg 5701 cttcattttg tagaacaaaa atgcaacgcg agagcgctaa tttttcaaac aaagaatctg 5761 agctgcattt ttacagaaca gaaatgcaac gcgaaagcgc tattttacca acgaagaatc 5821 tgtgcttcat ttttgtaaaa caaaaatgca acgcgagagc gctaattttt caaacaaaga 5881 atctgagctg catttttaca gaacagaaat gcaacgcgag agcgctattt taccaacaaa 5941 gaatctatac ttcttttttg ttctacaaaa atgcatcccg agagcgctat ttttctaaca 6001 aagcatctta gattactttt tttctccttt gtgcgctcta taatgcagtc tcttgataac 6061 tttttgcact gtaggtccgt taaggttaga agaaggctac tttggtgtct attttctctt 6121 ccataaaaaa agcctgactc cacttcccgc gtttactgat tactagcgaa gctgcgggtg 6181 cattttttca agataaaggc atccccgatt atattctata ccgatgtgga ttgcgcatac 6241 tttgtgaaca gaaagtgata gcgttgatga ttcttcattg gtcagaaaat tatgaacggt 6301 ttcttctatt ttgtctctat atactacgta taggaaatgt ttacattttc gtattgtttt 6361 cgattcactc tatgaatagt tcttactaca atttttttgt ctaaagagta atactagaga 6421 taaacataaa aaatgtagag gtcgagttta gatgcaagtt caaggagcga aaggtggatg 6481 ggtaggttat atagggatat agcacagaga tatatagcaa agagatactt ttgagcaatg 6541 tttgtggaag cggtattcgc aatattttag tagctcgtta cagtccggtg cgtttttggt 6601 tttttgaaag tgcgtcatca gagcgctttt ggttttcaaa agcgctctga agttcctata 6661 ctttctagct agagaatagg aacttcccga gcggccgcgt gttacaagct tgactccagc 6721 gtaactggac tgcaatcaac tcactggctc accttcacgg gtgggccttt cttcggtaga 6781 aaatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa 6841 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc 6901 cgaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta 6961 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct 7021 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg 7081 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag 7141 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc 7201 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg 7261 agagcgcacg agggagcttc caggggtaaa cgcctggtat ctttatagtc ctgtcgggtt 7321 tcgccacctc tgacttgagc atcgattttt gtgatgctcg tcaggggggc ggagcctatg 7381 gaaaaacgcc agcaacgcag aaaggcccac ccgaaggtga gccaggtgat tacatttggg 7441 ccctcatcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc 7501 atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt 7561 gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt 7621 tttttcctgt ttggtcactt accaatgctt aatcagtgag gcacctatct cagcgatctg 7681 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatgcggga 7741 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc 7801 agatttatca gcaataaacc agccagccgg gagggccgag cgcagaagtg atcctgcaac 7861 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc 7921 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc 7981 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc 8041 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgcca gaagtaagtt 8101 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc 8161 atccgtgaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg 8221 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag 8281 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgtaaac tctcaaggat 8341 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc 8401 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa 8461 aaagggaata agggcgacac ggaaatgttg aatactcatt ttagcttcct tagctcctga 8521 aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt 8581 ggaacctctt acgtgccgat caagtcaaaa gcctccggtc ggaggctttt gactttctgc 8641 tatggaggtc aggtatgatt taaatggtca gttaga //