LOCUS Exported 6303 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Yeast Expression plasmid for SARS-CoV-2 nsp10. ACCESSION . VERSION . KEYWORDS pGBW-m4046510 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6303) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 6303) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6303 /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 898..915 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" primer_bind complement(2058..2076) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 2427..3230 /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 3313..4295 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 4373..4402 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 4424..5011 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 5023..5050 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(5074..5265) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(5266..6126) /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 5889..5908 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6127..6229) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 6230..6261 /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 gcaggaaacg ctactgaagt tccagccaat tcaaccgtgt tgagcttttg tgctttcgcg 541 gtagatgctg ccaaggctta caaagactat cttgcttccg gtgggcaacc cattacgaac 601 tgcgtcaaga tgctgtgtac tcacacaggc accggtcagg ctataactgt taccccagag 661 gccaatatgg accaagaatc gttcggtggt gcatcttgtt gtctctactg ccgttgtcat 721 atcgatcacc caaaccctaa gggtttttgt gacttaaaag gtaagtacgt tcaaatccca 781 actacctgtg ctaacgatcc agtcggtttc accttgaaga atacagtttg tactgtatgc 841 ggtatgtgga aaggatacgg ttgttcttgc gatcaattga gagaacctat gttgcaactc 901 gtgccgagag gttctaaaat cgaggaaggt aagctagtca tttggataaa tggagataag 961 ggttacaacg gtttggcaga agtaggcaag aaatttgaaa aggacacagg tatcaaggtg 1021 acggttgaac atccagacaa gttagaagaa aaattccctc aagttgctgc tactggtgat 1081 ggtcccgaca ttattttctg ggcccacgat cgttttggtg gatatgctca gtcaggtttg 1141 ctggctgaga tcaccccaga caaggccttc caagataaat tgtacccatt cacttgggat 1201 gctgtcagat acaacggtaa gcttatagca tatccaattg ctgttgaagc cttatcgttg 1261 atctacaata aggacttgtt acctaaccca ccaaagacat gggaagaaat tccagctttg 1321 gataaagagc taaaggccaa gggaaaaagc gctttgatgt ttaacttaca agaaccttat 1381 ttcacctggc cattaatcgc tgctgacggt ggttacgcat ttaaatacga aaacggtaag 1441 tatgacatta aggatgtcgg cgttgataat gctggtgcaa aggctggttt gaccttcttg 1501 gttgatttaa taaagaataa acacatgaac gctgacactg actactctat cgccgaagcc 1561 gctttcaaca agggtgaaac tgctatgacc ataaatggtc catgggcctg gagtaacatt 1621 gatacttcta aagtaaacta tggtgtcact gttttgccta cattcaaagg ccaaccatcc 1681 aagccctttg tcggtgttct atccgctggc atcaatgctg cttctccaaa caaggagcta 1741 gctaaagaat ttttggaaaa ttacttgtta actgacgaag gtttggaggc cgttaacaag 1801 gacaagccat tgggtgctgt cgctttgaaa tcttacgaag aagaattggc taaggatccc 1861 agaattgccg ctaccatgga gaacgcccaa aagggtgaaa ttatgccaaa catcccacaa 1921 atgtcagcct tctggtacgc agttaggact gccgtgatta atgctgcttc cggtagacaa 1981 accgtcgacg aagcattaaa agatgctcaa acgtaaacag gccccttttc ctttgtcgat 2041 atcatgtaat tagttatgtc acgcttacat tcacgccctc ctcccacatc cgctctaacc 2101 gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt ttaatagtta 2161 tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg tacaaagcgg 2221 ccgccgaacc actagtgcac tgcagtacac ggtttccttg aaattttttt gattcggtaa 2281 tctccgaaca gaaggaagaa cgaaggaagg agcacagact tagattggta tatatacgca 2341 tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt aacccaactg cacagaacaa 2401 aaacgtgcag gaaacgaaga taaatcatgt cgaaagctac atataaggaa cgtgctgcta 2461 ctcatcctag tcctgttgct gccaagctat ttaatatcat gcacgaaaag caaacaaact 2521 tgtgtgcttc attggatgtt cgtaccacca aggaattact ggagttagtt gaagcattag 2581 gtcccaaaat ttgtttacta aaaacacatg tggatatctt gactgatttt tccatggagg 2641 gcacagttaa gccgctaaag gcattatccg ccaagtacaa ttttttactc ttcgaggaca 2701 gaaaatttgc tgacattggt aatacagtca aattgcagta ctctgcgggt gtatacagaa 2761 tagcagaatg ggcagacatt acgaatgcac acggtgtggt gggcccaggt attgttagcg 2821 gtttgaagca ggcggcagaa gaagtaacaa aggaacctag aggccttttg atgttagcag 2881 aattgtcatg caagggctcc ctatctactg gagaatatac taagggtact gttgacattg 2941 cgaagagcga caaagatttt gttatcggct ttattgctca aagagacatg ggtggaagag 3001 atgaaggtta cgattggttg attatgacac ccggtgtggg tttagatgac aagggagatg 3061 cattgggtca acagtataga accgtggatg atgtggtttc tacaggatct gacattatta 3121 ttgttggaag aggactattt gcaaagggaa gggatgctaa ggtagagggt gaacgttaca 3181 gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa aaaactgtat 3241 tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa ttatatcagt 3301 tattacccga gtaacgaagc atctgtgctt cattttgtag aacaaaaatg caacgcgaga 3361 gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg 3421 aaagcgctat tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa aaatgcaacg 3481 cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa cagaaatgca 3541 acgcgagagc gctattttac caacaaagaa tctatacttc ttttttgttc tacaaaaatg 3601 catcccgaga gcgctatttt tctaacaaag catcttagat tacttttttt ctcctttgtg 3661 cgctctataa tgcagtctct tgataacttt ttgcactgta ggtccgttaa ggttagaaga 3721 aggctacttt ggtgtctatt ttctcttcca taaaaaaagc ctgactccac ttcccgcgtt 3781 tactgattac tagcgaagct gcgggtgcat tttttcaaga taaaggcatc cccgattata 3841 ttctataccg atgtggattg cgcatacttt gtgaacagaa agtgatagcg ttgatgattc 3901 ttcattggtc agaaaattat gaacggtttc ttctattttg tctctatata ctacgtatag 3961 gaaatgttta cattttcgta ttgttttcga ttcactctat gaatagttct tactacaatt 4021 tttttgtcta aagagtaata ctagagataa acataaaaaa tgtagaggtc gagtttagat 4081 gcaagttcaa ggagcgaaag gtggatgggt aggttatata gggatatagc acagagatat 4141 atagcaaaga gatacttttg agcaatgttt gtggaagcgg tattcgcaat attttagtag 4201 ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc gtcatcagag cgcttttggt 4261 tttcaaaagc gctctgaagt tcctatactt tctagctaga gaataggaac ttcccgagcg 4321 gccgcgtgtt acaagcttga ctccagcgta actggactgc aatcaactca ctggctcacc 4381 ttcacgggtg ggcctttctt cggtagaaaa tcaaaggatc ttcttgagat cctttttttc 4441 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc 4501 cggatcaaga gctaccaact ctttttccga ggtaactggc ttcagcagag cgcagatacc 4561 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc 4621 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc 4681 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg 4741 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata 4801 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta 4861 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag gggtaaacgc 4921 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcatc gatttttgtg 4981 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcagaaa ggcccacccg 5041 aaggtgagcc aggtgattac atttgggccc tcatcagagg ttttcaccgt catcaccgaa 5101 acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc 5161 tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct 5221 gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcacttacc aatgcttaat 5281 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 5341 cgtcgtgtag ataactacga tgcgggaggg cttaccatct ggccccagtg ctgcaatgat 5401 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccgggag 5461 ggccgagcgc agaagtgatc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 5521 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 5581 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 5641 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 5701 tcctccgatc gttgccagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 5761 actgcataat tctcttactg tcatgccatc cgtgagatgc ttttctgtga ctggtgagta 5821 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 5881 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 5941 ttcttcgggg cgtaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 6001 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 6061 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 6121 actcatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag 6181 tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa gtcaaaagcc 6241 tccggtcgga ggcttttgac tttctgctat ggaggtcagg tatgatttaa atggtcagtt 6301 aga //