LOCUS Exported 7794 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Yeast Expression plasmid for SARS-CoV-2 nsp2. ACCESSION . VERSION . KEYWORDS pGBW-m4046385 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7794) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 7794) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7794 /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 2389..2406 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" primer_bind complement(3549..3567) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 3918..4721 /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 4804..5786 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 5864..5893 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 5915..6502 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 6514..6541 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(6565..6756) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(6757..7617) /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 7380..7399 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7618..7720) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 7721..7752 /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 tacactcgct atgtcgataa caacttctgt ggccctgatg gctaccctct tgagtgcatt 541 aaagaccttc tagcacgtgc tggtaaagct tcatgcactt tgtccgaaca actggacttt 601 attgacacta agaggggtgt atactgctgc cgtgaacatg agcatgaaat tgcttggtac 661 acggaacgtt ctgaaaagag ctatgaattg cagacacctt ttgaaattaa attggcaaag 721 aaatttgaca ccttcaatgg ggaatgtcca aattttgtat ttcccttaaa ttccataatc 781 aagactattc aaccaagggt tgaaaagaaa aagcttgatg gctttatggg tagaattcga 841 tctgtctatc cagttgcgtc accaaatgaa tgcaaccaaa tgtgcctttc aactctcatg 901 aagtgtgatc attgtggtga aacttcatgg cagacgggcg attttgttaa agccacttgc 961 gaattttgtg gcactgagaa tttgactaaa gaaggtgcca ctacttgtgg ttacttaccc 1021 caaaatgctg ttgttaaaat ttattgtcca gcatgtcaca attcagaagt aggacctgag 1081 catagtcttg ccgaatacca taatgaatct ggcttgaaaa ccattcttcg taagggtggt 1141 cgcactattg cctttggagg ctgtgtgttc tcttatgttg gttgccataa caagtgtgcc 1201 tattgggttc cacgtgctag cgctaacata ggttgtaacc atacaggtgt tgttggagaa 1261 ggttccgaag gtcttaatga caaccttctt gaaatactcc aaaaagagaa agtcaacatc 1321 aatattgttg gtgactttaa acttaatgaa gagatcgcca ttattttggc atctttttct 1381 gcttccacaa gtgcttttgt ggaaactgtg aaaggtttgg attataaagc attcaaacaa 1441 attgttgaat cctgtggtaa ttttaaagtt acaaaaggaa aagctaaaaa aggtgcctgg 1501 aatattggtg aacagaaatc aatactgagt cctctttatg catttgcatc agaggctgct 1561 cgtgttgtac gatcaatttt ctcccgcact cttgaaactg ctcaaaattc tgtgcgtgtt 1621 ttacagaagg ccgctataac aatactagat ggaatttcac agtattcact gagactcatt 1681 gatgctatga tgttcacatc tgatttggct actaacaatc tagttgtaat ggcctacatt 1741 acaggtggtg ttgttcagtt gacttcgcag tggctaacta acatctttgg cactgtttat 1801 gaaaaactca aacccgtcct tgattggctt gaagagaagt ttaaggaagg tgtagagttt 1861 cttagagacg gttgggaaat tgttaaattt atctcaacct gtgcttgtga aattgtcggt 1921 ggacaaattg tcacctgtgc aaaggaaatt aaggagagtg ttcagacatt ctttaagctt 1981 gtaaataaat ttttggcttt gtgtgctgac tctatcatta ttggtggagc taaacttaaa 2041 gccttgaatt taggtgaaac atttgtcacg cactcaaagg gattgtacag aaagtgtgtt 2101 aaatccagag aagaaactgg cctactcatg cctctaaaag ccccaaaaga aattatcttc 2161 ttagagggag aaacacttcc cacagaagtg ttaacagagg aagttgtctt gaaaactggt 2221 gatttacaac cattagaaca acctactagt gaagctgttg aagctccatt ggttggtaca 2281 ccagtttgta ttaacgggct tatgttgctc gaaatcaaag acacagaaaa gtactgtgcc 2341 cttgcaccta atatgatggt aacaaacaat accttcacac tcaaaggcct cgtgccgaga 2401 ggttctaaaa tcgaggaagg taagctagtc atttggataa atggagataa gggttacaac 2461 ggtttggcag aagtaggcaa gaaatttgaa aaggacacag gtatcaaggt gacggttgaa 2521 catccagaca agttagaaga aaaattccct caagttgctg ctactggtga tggtcccgac 2581 attattttct gggcccacga tcgttttggt ggatatgctc agtcaggttt gctggctgag 2641 atcaccccag acaaggcctt ccaagataaa ttgtacccat tcacttggga tgctgtcaga 2701 tacaacggta agcttatagc atatccaatt gctgttgaag ccttatcgtt gatctacaat 2761 aaggacttgt tacctaaccc accaaagaca tgggaagaaa ttccagcttt ggataaagag 2821 ctaaaggcca agggaaaaag cgctttgatg tttaacttac aagaacctta tttcacctgg 2881 ccattaatcg ctgctgacgg tggttacgca tttaaatacg aaaacggtaa gtatgacatt 2941 aaggatgtcg gcgttgataa tgctggtgca aaggctggtt tgaccttctt ggttgattta 3001 ataaagaata aacacatgaa cgctgacact gactactcta tcgccgaagc cgctttcaac 3061 aagggtgaaa ctgctatgac cataaatggt ccatgggcct ggagtaacat tgatacttct 3121 aaagtaaact atggtgtcac tgttttgcct acattcaaag gccaaccatc caagcccttt 3181 gtcggtgttc tatccgctgg catcaatgct gcttctccaa acaaggagct agctaaagaa 3241 tttttggaaa attacttgtt aactgacgaa ggtttggagg ccgttaacaa ggacaagcca 3301 ttgggtgctg tcgctttgaa atcttacgaa gaagaattgg ctaaggatcc cagaattgcc 3361 gctaccatgg agaacgccca aaagggtgaa attatgccaa acatcccaca aatgtcagcc 3421 ttctggtacg cagttaggac tgccgtgatt aatgctgctt ccggtagaca aaccgtcgac 3481 gaagcattaa aagatgctca aacgtaaaca ggcccctttt cctttgtcga tatcatgtaa 3541 ttagttatgt cacgcttaca ttcacgccct cctcccacat ccgctctaac cgaaaaggaa 3601 ggagttagac aacctgaagt ctaggtccct atttattttt tttaatagtt atgttagtat 3661 taagaacgtt atttatattt caaatttttc ttttttttct gtacaaagcg gccgccgaac 3721 cactagtgca ctgcagtaca cggtttcctt gaaatttttt tgattcggta atctccgaac 3781 agaaggaaga acgaaggaag gagcacagac ttagattggt atatatacgc atatgtagtg 3841 ttgaagaaac atgaaattgc ccagtattct taacccaact gcacagaaca aaaacgtgca 3901 ggaaacgaag ataaatcatg tcgaaagcta catataagga acgtgctgct actcatccta 3961 gtcctgttgc tgccaagcta tttaatatca tgcacgaaaa gcaaacaaac ttgtgtgctt 4021 cattggatgt tcgtaccacc aaggaattac tggagttagt tgaagcatta ggtcccaaaa 4081 tttgtttact aaaaacacat gtggatatct tgactgattt ttccatggag ggcacagtta 4141 agccgctaaa ggcattatcc gccaagtaca attttttact cttcgaggac agaaaatttg 4201 ctgacattgg taatacagtc aaattgcagt actctgcggg tgtatacaga atagcagaat 4261 gggcagacat tacgaatgca cacggtgtgg tgggcccagg tattgttagc ggtttgaagc 4321 aggcggcaga agaagtaaca aaggaaccta gaggcctttt gatgttagca gaattgtcat 4381 gcaagggctc cctatctact ggagaatata ctaagggtac tgttgacatt gcgaagagcg 4441 acaaagattt tgttatcggc tttattgctc aaagagacat gggtggaaga gatgaaggtt 4501 acgattggtt gattatgaca cccggtgtgg gtttagatga caagggagat gcattgggtc 4561 aacagtatag aaccgtggat gatgtggttt ctacaggatc tgacattatt attgttggaa 4621 gaggactatt tgcaaaggga agggatgcta aggtagaggg tgaacgttac agaaaagcag 4681 gctgggaagc atatttgaga agatgcggcc agcaaaacta aaaaactgta ttataagtaa 4741 atgcatgtat actaaactca caaattagag cttcaattta attatatcag ttattacccg 4801 agtaacgaag catctgtgct tcattttgta gaacaaaaat gcaacgcgag agcgctaatt 4861 tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc gaaagcgcta 4921 ttttaccaac gaagaatctg tgcttcattt ttgtaaaaca aaaatgcaac gcgagagcgc 4981 taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgagag 5041 cgctatttta ccaacaaaga atctatactt cttttttgtt ctacaaaaat gcatcccgag 5101 agcgctattt ttctaacaaa gcatcttaga ttactttttt tctcctttgt gcgctctata 5161 atgcagtctc ttgataactt tttgcactgt aggtccgtta aggttagaag aaggctactt 5221 tggtgtctat tttctcttcc ataaaaaaag cctgactcca cttcccgcgt ttactgatta 5281 ctagcgaagc tgcgggtgca ttttttcaag ataaaggcat ccccgattat attctatacc 5341 gatgtggatt gcgcatactt tgtgaacaga aagtgatagc gttgatgatt cttcattggt 5401 cagaaaatta tgaacggttt cttctatttt gtctctatat actacgtata ggaaatgttt 5461 acattttcgt attgttttcg attcactcta tgaatagttc ttactacaat ttttttgtct 5521 aaagagtaat actagagata aacataaaaa atgtagaggt cgagtttaga tgcaagttca 5581 aggagcgaaa ggtggatggg taggttatat agggatatag cacagagata tatagcaaag 5641 agatactttt gagcaatgtt tgtggaagcg gtattcgcaa tattttagta gctcgttaca 5701 gtccggtgcg tttttggttt tttgaaagtg cgtcatcaga gcgcttttgg ttttcaaaag 5761 cgctctgaag ttcctatact ttctagctag agaataggaa cttcccgagc ggccgcgtgt 5821 tacaagcttg actccagcgt aactggactg caatcaactc actggctcac cttcacgggt 5881 gggcctttct tcggtagaaa atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa 5941 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag 6001 agctaccaac tctttttccg aggtaactgg cttcagcaga gcgcagatac caaatactgt 6061 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata 6121 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac 6181 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg 6241 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg 6301 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag 6361 cggcagggtc ggaacaggag agcgcacgag ggagcttcca ggggtaaacg cctggtatct 6421 ttatagtcct gtcgggtttc gccacctctg acttgagcat cgatttttgt gatgctcgtc 6481 aggggggcgg agcctatgga aaaacgccag caacgcagaa aggcccaccc gaaggtgagc 6541 caggtgatta catttgggcc ctcatcagag gttttcaccg tcatcaccga aacgcgcgag 6601 gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc 6661 atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg 6721 ggccatgtta agggcggttt tttcctgttt ggtcacttac caatgcttaa tcagtgaggc 6781 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta 6841 gataactacg atgcgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga 6901 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggga gggccgagcg 6961 cagaagtgat cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc 7021 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat 7081 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag 7141 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat 7201 cgttgccaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa 7261 ttctcttact gtcatgccat ccgtgagatg cttttctgtg actggtgagt actcaaccaa 7321 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga 7381 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg 7441 gcgtaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc 7501 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg 7561 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatttt 7621 agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat 7681 ttcattatgg tgaaagttgg aacctcttac gtgccgatca agtcaaaagc ctccggtcgg 7741 aggcttttga ctttctgcta tggaggtcag gtatgattta aatggtcagt taga //