LOCUS Exported 10605 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Yeast expression plasmid for SARS-CoV-2 nsp3. ACCESSION . VERSION . KEYWORDS pGBW-m4046663 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10605) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 10605) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10605 /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" primer_bind complement(6360..6378) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 6729..7532 /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 7615..8597 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 8675..8704 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 8726..9313 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 9325..9352 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(9376..9567) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(9568..10428) /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 10191..10210 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(10429..10531) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 10532..10563 /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 gcccccacta aggtcacatt cggggatgac accgtgatag aggttcaggg ttacaaatcc 541 gttaacatca cttttgaatt agatgaaaga attgacaagg tattgaatga aaagtgctca 601 gcatataccg ttgaactggg tacggaggtc aacgaatttg cttgtgtcgt tgcggatgct 661 gttatcaaaa ctttgcaacc tgtgtctgaa cttctaaccc cattaggaat tgacttggat 721 gaatggtcta tggctactta ctatctattc gatgaaagtg gtgagttcaa gttggcttcc 781 cacatgtact gttcgttcta cccaccagac gaagacgaag aagaaggcga ctgtgaagaa 841 gaagagtttg aaccaagcac ccaatatgaa tacggtacgg aagatgatta ccaaggtaag 901 ccattggaat ttggtgccac ttctgctgca ttgcaacctg aagaagagca agaagaagat 961 tggttagacg atgactctca gcaaacagtc ggtcaacaag acggttccga ggataatcaa 1021 actaccacca ttcaaacaat cgttgaggta caaccacaac ttgaaatgga attgactcca 1081 gtcgttcaga ccatagaagt taactcgttc tcaggttact tgaagctaac tgataatgtt 1141 tatattaaaa acgctgacat cgtcgaagaa gcaaagaaag ttaagcctac cgtcgtggtt 1201 aacgccgcta acgtttattt gaagcatggt ggcggtgtag ctggtgcttt aaacaaagcc 1261 actaataacg ctatgcaagt cgaatctgac gattacattg ctaccaatgg tccattgaag 1321 gtgggaggct cttgcgtctt aagtggacac aacttggcga aacattgtct ccacgttgtt 1381 ggtcccaatg ttaacaaggg tgaagacatt caactgttga agtccgccta cgaaaatttc 1441 aaccaacatg aggttctctt ggcaccatta ctgagtgctg gtatcttcgg cgctgaccca 1501 attcactctt taagggtctg tgtggatact gtaagaacaa acgtctactt ggccgttttc 1561 gataaaaacc tttatgacaa gttagtctct tcctttttgg aaatgaagtc agaaaagcaa 1621 gttgaacaga aaatcgcgga aatccctaag gaagaagtaa agccgtttat aaccgaatcg 1681 aagccctctg ttgagcaaag aaagcaagat gacaaaaaga ttaaagcttg tgttgaggaa 1741 gtaacaacta cattagagga aactaagttt ctcacggaaa atctattgtt gtacatagat 1801 atcaatggta acttgcaccc agactccgct actttggtca gcgatatcga cattacattc 1861 ttgaagaagg acgctccata tattgtgggt gacgttgtcc aagaaggtgt tttaactgct 1921 gttgtcatcc caactaagaa ggcagggggt accacggaga tgctagctaa ggccttgcgt 1981 aaagtcccga ctgataatta tattaccacc taccctggtc agggtctaaa cggttacacg 2041 gttgaagaag ctaaaactgt tctaaaaaag tgtaagtctg ctttttacat attgccatca 2101 attatttcca acgaaaagca agaaatattg ggtaccgtat cttggaattt gagagaaatg 2161 ttagctcatg ctgaggaaac tagaaagttg atgcctgtct gcgttgaaac taaagctatc 2221 gtcagcacga tccaaagaaa atacaagggt attaaaatcc aagaaggcgt tgtggactat 2281 ggagcccgtt tctatttcta cacttcgaag accaccgtcg cttcgttgat caacaccttg 2341 aacgatctca acgagacact tgttactatg ccactcggct atgtaactca cggtcttaac 2401 ttggaagaag ctgcgagata catgcgctct ctgaaagttc ctgccacagt ctccgtatct 2461 tcaccagacg ctgttacagc ctataatgga tacttgactt cttcatcaaa aaccccagaa 2521 gaacatttca tcgaaactat ctctctagct ggtagttaca aggattggtc ttactccggt 2581 caatccacac aactaggtat agagtttctg aagagaggtg acaagagtgt ttactacacc 2641 agcaatccaa cgacctttca tttagatggt gaagtcatta cgttcgataa cttgaagact 2701 ttgttgagtt tgcgggaagt ccgaactatc aaggttttca ctaccgttga taacattaac 2761 ctccacaccc aagttgttga tatgtctatg acttatggtc aacagttcgg tccgacatat 2821 ttggatggtg ctgacgttac taaaattaag ccacacaact cccacgaagg gaagacattc 2881 tacgtgcttc caaatgacga cactttacga gtcgaagcct ttgagtacta ccataccacg 2941 gacccatctt tccttggtag atacatgtcc gctctgaatc acactaaaaa gtggaaatac 3001 ccacaagtta acggtttgac ttctattaag tgggccgata ataactgtta cttggctacc 3061 gccttattga ctttgcaaca gatagaactc aagtttaatc ctcccgctct acaagatgca 3121 tattatagag ctagagcagg cgaagcggct aatttctgtg ctttaatttt ggcttactgt 3181 aacaagaccg taggtgaatt gggggatgtc agagagacta tgtcctatct tttccagcac 3241 gcaaacttgg attcatgtaa gagagtgtta aatgtagtct gcaagacctg cggtcaacaa 3301 caaactacct taaagggtgt tgaagccgtt atgtacatgg gtacactttc ttacgaacaa 3361 tttaagaagg gtgtccaaat cccatgtact tgtggtaagc aagccactaa atacttggtc 3421 cagcaagaaa gccctttcgt tatgatgagc gccccaccag ctcaatacga gctgaagcat 3481 ggaactttca cctgtgcttc tgaatatacc ggtaactacc aatgcggtca ctacaagcat 3541 attactagca aagaaacttt gtactgtatc gatggtgcct tgttgactaa gtcctcagaa 3601 tataagggcc caattactga cgtgttttac aaagagaata gttacactac cacaattaag 3661 ccagtaacat acaaactgga tggtgttgtg tgtaccgaga tcgaccctaa gttagataac 3721 tattacaaga aggacaactc ttactttacc gaacaaccga tcgacctagt cccaaatcag 3781 ccatatccaa atgcttcttt tgataacttc aagttcgttt gtgataacat aaaattcgcc 3841 gacgatttaa accaattgac cggttacaaa aagccggctt caagagagtt gaaagtcact 3901 ttcttcccag atttaaatgg cgatgtcgtt gctattgact ataagcacta cactccatct 3961 ttcaagaagg gcgccaagtt gctacacaag ccaattgtct ggcacgttaa caacgctacc 4021 aacaaggcta cgtataagcc taatacctgg tgtatcagat gtctatggtc caccaaacca 4081 gttgaaacga gtaactcttt tgacgttctg aaatcagaag atgcccaagg gatggataac 4141 ttagcatgtg aagacttgaa acctgtttca gaagaagttg tggaaaatcc aactattcag 4201 aaggacgtgt tggaatgcaa cgtgaaaaca actgaagtag ttggtgatat aatcctgaag 4261 ccagctaaca actcactgaa gattaccgaa gaagtcggtc acaccgactt gatggcagcc 4321 tacgttgata actcttccct cacgatcaaa aagccaaatg aattgtctcg tgtactcggg 4381 ctaaagaccc tagctaccca tggtctagcg gctgtcaact ccgtgccgtg ggacaccatc 4441 gctaactacg ctaagccttt tttgaataag gtcgttagca ccacgacaaa tatagttact 4501 agatgtttga acagagtgtg taccaattac atgccttatt tctttactct attgctgcaa 4561 ttgtgcactt tcactcgctc tacaaactcc agaattaaag catctatgcc aaccaccatc 4621 gccaagaaca cggttaagtc agtaggtaag ttctgcttag aggcaagctt taactatttg 4681 aagtctccaa acttctctaa gttgatcaat attattatct ggtttttact tttgagcgtt 4741 tgtttgggtt ccttgatcta ctcaacggct gctttgggtg tcttgatgtc taacttgggt 4801 atgccgagtt actgtactgg ttatagagaa ggttatttga actcaacaaa cgttactatt 4861 gctacgtact gcaccggttc cattccatgt tctgtttgtt tatccggttt agactcgttg 4921 gacacttacc catctttaga aaccatccaa attaccattt cctctttcaa gtgggatttg 4981 acagctttcg gtcttgtcgc tgaatggttt ttggcctata tattgtttac tcgtttcttc 5041 tacgtcttag gcttggccgc tatcatgcag ttgtttttct cttattttgc agtccatttt 5101 atctcaaatt cgtggttgat gtggttgatt attaatctcg ttcaaatggc tccaatttcg 5161 gcaatggtta ggatgtacat tttctttgct tcattctatt atgtatggaa atcttacgtc 5221 catgtggttg atggttgtaa ctcctcgacc tgtatgatgt gttacaagag aaatagagcg 5281 actagggttg agtgcacaac tatcgttaac ggtgtcagac gttctttcta cgtatacgct 5341 aacggtggta agggcttttg taaactccat aactggaatt gtgttaattg tgataccttt 5401 tgcgctggtt ctacctttat tagtgatgaa gtagccagag acttaagtct ccaattcaaa 5461 cggccaatca acccaactga tcaatcctct tatattgtcg atagtgtcac cgttaagaac 5521 ggtagtatcc acttatactt cgataaggcc ggtcaaaaga catacgaaag acactcattg 5581 tcccactttg ttaatttaga caatttgaga gctaataata ctaaggggtc tttacccatc 5641 aacgtcatag tcttcgatgg taagtctaaa tgtgaagaat cctccgctaa gtctgcgtca 5701 gtttactact ctcaattgat gtgccaaccc atcctacttc tggatcaagc tttggtatct 5761 gacgttggtg actctgccga ggttgctgtt aagatgttcg atgcttacgt taacacattt 5821 tcatcaactt tcaacgtgcc aatggaaaag ctgaagactt tggtggctac cgctgaagca 5881 gaattggcta agaatgtttc tttggataac gttttaagta cgttcatttc cgcagctcgt 5941 cagggtttcg tcgactctga cgtagaaacc aaagacgtcg tcgaatgttt gaagctctct 6001 catcaatccg atattgaagt cactggtgac tcctgtaata actacatgct gacgtataac 6061 aaagtggaaa acatgactcc tagagatttg ggagcgtgca tcgactgttc tgctagacac 6121 ataaacgctc aagttgcaaa atcccataat attgctttga tctggaacgt taaagatttc 6181 atgtcactat ccgagcaact taggaaacaa atccggtctg cggcaaagaa aaataacctt 6241 ccattcaaat taacatgtgc aacaactaga caagttgtga acgttgttac tactaagatc 6301 gctctgaagg gtggttaaac aggccccttt tcctttgtcg atatcatgta attagttatg 6361 tcacgcttac attcacgccc tcctcccaca tccgctctaa ccgaaaagga aggagttaga 6421 caacctgaag tctaggtccc tatttatttt ttttaatagt tatgttagta ttaagaacgt 6481 tatttatatt tcaaattttt cttttttttc tgtacaaagc ggccgccgaa ccactagtgc 6541 actgcagtac acggtttcct tgaaattttt ttgattcggt aatctccgaa cagaaggaag 6601 aacgaaggaa ggagcacaga cttagattgg tatatatacg catatgtagt gttgaagaaa 6661 catgaaattg cccagtattc ttaacccaac tgcacagaac aaaaacgtgc aggaaacgaa 6721 gataaatcat gtcgaaagct acatataagg aacgtgctgc tactcatcct agtcctgttg 6781 ctgccaagct atttaatatc atgcacgaaa agcaaacaaa cttgtgtgct tcattggatg 6841 ttcgtaccac caaggaatta ctggagttag ttgaagcatt aggtcccaaa atttgtttac 6901 taaaaacaca tgtggatatc ttgactgatt tttccatgga gggcacagtt aagccgctaa 6961 aggcattatc cgccaagtac aattttttac tcttcgagga cagaaaattt gctgacattg 7021 gtaatacagt caaattgcag tactctgcgg gtgtatacag aatagcagaa tgggcagaca 7081 ttacgaatgc acacggtgtg gtgggcccag gtattgttag cggtttgaag caggcggcag 7141 aagaagtaac aaaggaacct agaggccttt tgatgttagc agaattgtca tgcaagggct 7201 ccctatctac tggagaatat actaagggta ctgttgacat tgcgaagagc gacaaagatt 7261 ttgttatcgg ctttattgct caaagagaca tgggtggaag agatgaaggt tacgattggt 7321 tgattatgac acccggtgtg ggtttagatg acaagggaga tgcattgggt caacagtata 7381 gaaccgtgga tgatgtggtt tctacaggat ctgacattat tattgttgga agaggactat 7441 ttgcaaaggg aagggatgct aaggtagagg gtgaacgtta cagaaaagca ggctgggaag 7501 catatttgag aagatgcggc cagcaaaact aaaaaactgt attataagta aatgcatgta 7561 tactaaactc acaaattaga gcttcaattt aattatatca gttattaccc gagtaacgaa 7621 gcatctgtgc ttcattttgt agaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca 7681 aagaatctga gctgcatttt tacagaacag aaatgcaacg cgaaagcgct attttaccaa 7741 cgaagaatct gtgcttcatt tttgtaaaac aaaaatgcaa cgcgagagcg ctaatttttc 7801 aaacaaagaa tctgagctgc atttttacag aacagaaatg caacgcgaga gcgctatttt 7861 accaacaaag aatctatact tcttttttgt tctacaaaaa tgcatcccga gagcgctatt 7921 tttctaacaa agcatcttag attacttttt ttctcctttg tgcgctctat aatgcagtct 7981 cttgataact ttttgcactg taggtccgtt aaggttagaa gaaggctact ttggtgtcta 8041 ttttctcttc cataaaaaaa gcctgactcc acttcccgcg tttactgatt actagcgaag 8101 ctgcgggtgc attttttcaa gataaaggca tccccgatta tattctatac cgatgtggat 8161 tgcgcatact ttgtgaacag aaagtgatag cgttgatgat tcttcattgg tcagaaaatt 8221 atgaacggtt tcttctattt tgtctctata tactacgtat aggaaatgtt tacattttcg 8281 tattgttttc gattcactct atgaatagtt cttactacaa tttttttgtc taaagagtaa 8341 tactagagat aaacataaaa aatgtagagg tcgagtttag atgcaagttc aaggagcgaa 8401 aggtggatgg gtaggttata tagggatata gcacagagat atatagcaaa gagatacttt 8461 tgagcaatgt ttgtggaagc ggtattcgca atattttagt agctcgttac agtccggtgc 8521 gtttttggtt ttttgaaagt gcgtcatcag agcgcttttg gttttcaaaa gcgctctgaa 8581 gttcctatac tttctagcta gagaatagga acttcccgag cggccgcgtg ttacaagctt 8641 gactccagcg taactggact gcaatcaact cactggctca ccttcacggg tgggcctttc 8701 ttcggtagaa aatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct 8761 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa 8821 ctctttttcc gaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt 8881 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct 8941 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga 9001 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac 9061 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg 9121 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt 9181 cggaacagga gagcgcacga gggagcttcc aggggtaaac gcctggtatc tttatagtcc 9241 tgtcgggttt cgccacctct gacttgagca tcgatttttg tgatgctcgt caggggggcg 9301 gagcctatgg aaaaacgcca gcaacgcaga aaggcccacc cgaaggtgag ccaggtgatt 9361 acatttgggc cctcatcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg 9421 gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc 9481 cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt 9541 aagggcggtt ttttcctgtt tggtcactta ccaatgctta atcagtgagg cacctatctc 9601 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac 9661 gatgcgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc 9721 accggctcca gatttatcag caataaacca gccagccggg agggccgagc gcagaagtga 9781 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag 9841 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc 9901 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac 9961 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgccag 10021 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac 10081 tgtcatgcca tccgtgagat gcttttctgt gactggtgag tactcaacca agtcattctg 10141 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc 10201 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgtaaact 10261 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg 10321 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa 10381 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcattt tagcttcctt 10441 agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta tttcattatg 10501 gtgaaagttg gaacctctta cgtgccgatc aagtcaaaag cctccggtcg gaggcttttg 10561 actttctgct atggaggtca ggtatgattt aaatggtcag ttaga //