LOCUS Exported 5103 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Yeast Expression plasmid for SARS-CoV-2 nsp9. ACCESSION . VERSION . KEYWORDS pGBW-m4046635 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5103) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 5103) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5103 /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(858..876) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 1227..2030 /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 2113..3095 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 3173..3202 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 3224..3811 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 3823..3850 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(3874..4065) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(4066..4926) /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 4689..4708 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4927..5029) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 5030..5061 /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 aatgagctta gtcctgttgc actacgacag atgtcttgtg ctgccggtac tacacaaact 541 gcttgcactg atgacaatgc gttagcttac tacaacacaa caaagggagg taggtttgta 601 cttgcactgt tatccgattt acaggatttg aaatgggcta gattccctaa gagtgatgga 661 actggtacta tctatacaga actggaacca ccttgtaggt ttgttacaga cacacctaaa 721 ggtcctaaag tgaagtattt atactttatt aaaggattaa acaacctaaa tagaggtatg 781 gtacttggta gtttagctgc cacagtacgt ctataaacag gccccttttc ctttgtcgat 841 atcatgtaat tagttatgtc acgcttacat tcacgccctc ctcccacatc cgctctaacc 901 gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt ttaatagtta 961 tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg tacaaagcgg 1021 ccgccgaacc actagtgcac tgcagtacac ggtttccttg aaattttttt gattcggtaa 1081 tctccgaaca gaaggaagaa cgaaggaagg agcacagact tagattggta tatatacgca 1141 tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt aacccaactg cacagaacaa 1201 aaacgtgcag gaaacgaaga taaatcatgt cgaaagctac atataaggaa cgtgctgcta 1261 ctcatcctag tcctgttgct gccaagctat ttaatatcat gcacgaaaag caaacaaact 1321 tgtgtgcttc attggatgtt cgtaccacca aggaattact ggagttagtt gaagcattag 1381 gtcccaaaat ttgtttacta aaaacacatg tggatatctt gactgatttt tccatggagg 1441 gcacagttaa gccgctaaag gcattatccg ccaagtacaa ttttttactc ttcgaggaca 1501 gaaaatttgc tgacattggt aatacagtca aattgcagta ctctgcgggt gtatacagaa 1561 tagcagaatg ggcagacatt acgaatgcac acggtgtggt gggcccaggt attgttagcg 1621 gtttgaagca ggcggcagaa gaagtaacaa aggaacctag aggccttttg atgttagcag 1681 aattgtcatg caagggctcc ctatctactg gagaatatac taagggtact gttgacattg 1741 cgaagagcga caaagatttt gttatcggct ttattgctca aagagacatg ggtggaagag 1801 atgaaggtta cgattggttg attatgacac ccggtgtggg tttagatgac aagggagatg 1861 cattgggtca acagtataga accgtggatg atgtggtttc tacaggatct gacattatta 1921 ttgttggaag aggactattt gcaaagggaa gggatgctaa ggtagagggt gaacgttaca 1981 gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa aaaactgtat 2041 tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa ttatatcagt 2101 tattacccga gtaacgaagc atctgtgctt cattttgtag aacaaaaatg caacgcgaga 2161 gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg 2221 aaagcgctat tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa aaatgcaacg 2281 cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa cagaaatgca 2341 acgcgagagc gctattttac caacaaagaa tctatacttc ttttttgttc tacaaaaatg 2401 catcccgaga gcgctatttt tctaacaaag catcttagat tacttttttt ctcctttgtg 2461 cgctctataa tgcagtctct tgataacttt ttgcactgta ggtccgttaa ggttagaaga 2521 aggctacttt ggtgtctatt ttctcttcca taaaaaaagc ctgactccac ttcccgcgtt 2581 tactgattac tagcgaagct gcgggtgcat tttttcaaga taaaggcatc cccgattata 2641 ttctataccg atgtggattg cgcatacttt gtgaacagaa agtgatagcg ttgatgattc 2701 ttcattggtc agaaaattat gaacggtttc ttctattttg tctctatata ctacgtatag 2761 gaaatgttta cattttcgta ttgttttcga ttcactctat gaatagttct tactacaatt 2821 tttttgtcta aagagtaata ctagagataa acataaaaaa tgtagaggtc gagtttagat 2881 gcaagttcaa ggagcgaaag gtggatgggt aggttatata gggatatagc acagagatat 2941 atagcaaaga gatacttttg agcaatgttt gtggaagcgg tattcgcaat attttagtag 3001 ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc gtcatcagag cgcttttggt 3061 tttcaaaagc gctctgaagt tcctatactt tctagctaga gaataggaac ttcccgagcg 3121 gccgcgtgtt acaagcttga ctccagcgta actggactgc aatcaactca ctggctcacc 3181 ttcacgggtg ggcctttctt cggtagaaaa tcaaaggatc ttcttgagat cctttttttc 3241 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc 3301 cggatcaaga gctaccaact ctttttccga ggtaactggc ttcagcagag cgcagatacc 3361 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc 3421 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc 3481 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg 3541 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata 3601 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta 3661 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag gggtaaacgc 3721 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcatc gatttttgtg 3781 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcagaaa ggcccacccg 3841 aaggtgagcc aggtgattac atttgggccc tcatcagagg ttttcaccgt catcaccgaa 3901 acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc 3961 tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct 4021 gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcacttacc aatgcttaat 4081 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 4141 cgtcgtgtag ataactacga tgcgggaggg cttaccatct ggccccagtg ctgcaatgat 4201 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccgggag 4261 ggccgagcgc agaagtgatc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 4321 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 4381 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 4441 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 4501 tcctccgatc gttgccagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 4561 actgcataat tctcttactg tcatgccatc cgtgagatgc ttttctgtga ctggtgagta 4621 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 4681 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 4741 ttcttcgggg cgtaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 4801 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 4861 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 4921 actcatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag 4981 tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa gtcaaaagcc 5041 tccggtcgga ggcttttgac tttctgctat ggaggtcagg tatgatttaa atggtcagtt 5101 aga //