LOCUS Exported 5433 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Yeast Expression plasmid for SARS-CoV-2 M (membrane). ACCESSION . VERSION . KEYWORDS pGBW-m4046688 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5433) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 5433) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5433 /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(1188..1206) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 1557..2360 /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 2443..3425 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 3503..3532 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 3554..4141 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 4153..4180 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(4204..4395) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(4396..5256) /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 5019..5038 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5257..5359) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 5360..5391 /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 gctgattcca atggcaccat cacagtagaa gagctgaaaa agctccttga acaatggaac 541 ttggttatag gttttttatt cttgacttgg atttgcctat tgcagttcgc ctatgcaaac 601 agaaaccgtt ttttgtacat tatcaagtta attttcttgt ggctgttatg gcctgtcact 661 cttgcttgtt ttgttttggc tgccgtctac cggataaatt ggatcaccgg tggtatcgct 721 attgcaatgg cttgtttggt tggattgatg tggttgagtt atttcattgc ctcattcaga 781 ttattcgcta ggactagatc gatgtggtcc tttaatccag aaacgaacat cttgttgaac 841 gtgccactcc atggtactat tttgaccaga ccgctattag aatctgagtt ggttatcggt 901 gctgtcattc taagagggca cttacgtatc gcgggtcacc atttgggaag atgtgacatt 961 aaggatcttc caaaagaaat cacagtcgct acctctcgaa ctctatctta ctacaagttg 1021 ggtgcctcac aaagggttgc tggtgacagc ggtttcgctg cctattctcg ctaccgtatc 1081 ggcaactaca aattaaatac cgatcactcc tctagttctg acaatattgc cttattggtg 1141 caataaacag gccccttttc ctttgtcgat atcatgtaat tagttatgtc acgcttacat 1201 tcacgccctc ctcccacatc cgctctaacc gaaaaggaag gagttagaca acctgaagtc 1261 taggtcccta tttatttttt ttaatagtta tgttagtatt aagaacgtta tttatatttc 1321 aaatttttct tttttttctg tacaaagcgg ccgccgaacc actagtgcac tgcagtacac 1381 ggtttccttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg 1441 agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc 1501 cagtattctt aacccaactg cacagaacaa aaacgtgcag gaaacgaaga taaatcatgt 1561 cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct gccaagctat 1621 ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt cgtaccacca 1681 aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta aaaacacatg 1741 tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag gcattatccg 1801 ccaagtacaa ttttttactc ttcgaggaca gaaaatttgc tgacattggt aatacagtca 1861 aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt acgaatgcac 1921 acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa gaagtaacaa 1981 aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc ctatctactg 2041 gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt gttatcggct 2101 ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg attatgacac 2161 ccggtgtggg tttagatgac aagggagatg cattgggtca acagtataga accgtggatg 2221 atgtggtttc tacaggatct gacattatta ttgttggaag aggactattt gcaaagggaa 2281 gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca tatttgagaa 2341 gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata ctaaactcac 2401 aaattagagc ttcaatttaa ttatatcagt tattacccga gtaacgaagc atctgtgctt 2461 cattttgtag aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc 2521 tgcattttta cagaacagaa atgcaacgcg aaagcgctat tttaccaacg aagaatctgt 2581 gcttcatttt tgtaaaacaa aaatgcaacg cgagagcgct aatttttcaa acaaagaatc 2641 tgagctgcat ttttacagaa cagaaatgca acgcgagagc gctattttac caacaaagaa 2701 tctatacttc ttttttgttc tacaaaaatg catcccgaga gcgctatttt tctaacaaag 2761 catcttagat tacttttttt ctcctttgtg cgctctataa tgcagtctct tgataacttt 2821 ttgcactgta ggtccgttaa ggttagaaga aggctacttt ggtgtctatt ttctcttcca 2881 taaaaaaagc ctgactccac ttcccgcgtt tactgattac tagcgaagct gcgggtgcat 2941 tttttcaaga taaaggcatc cccgattata ttctataccg atgtggattg cgcatacttt 3001 gtgaacagaa agtgatagcg ttgatgattc ttcattggtc agaaaattat gaacggtttc 3061 ttctattttg tctctatata ctacgtatag gaaatgttta cattttcgta ttgttttcga 3121 ttcactctat gaatagttct tactacaatt tttttgtcta aagagtaata ctagagataa 3181 acataaaaaa tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag gtggatgggt 3241 aggttatata gggatatagc acagagatat atagcaaaga gatacttttg agcaatgttt 3301 gtggaagcgg tattcgcaat attttagtag ctcgttacag tccggtgcgt ttttggtttt 3361 ttgaaagtgc gtcatcagag cgcttttggt tttcaaaagc gctctgaagt tcctatactt 3421 tctagctaga gaataggaac ttcccgagcg gccgcgtgtt acaagcttga ctccagcgta 3481 actggactgc aatcaactca ctggctcacc ttcacgggtg ggcctttctt cggtagaaaa 3541 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa 3601 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga 3661 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt 3721 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt 3781 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata 3841 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt 3901 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac 3961 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga 4021 gcgcacgagg gagcttccag gggtaaacgc ctggtatctt tatagtcctg tcgggtttcg 4081 ccacctctga cttgagcatc gatttttgtg atgctcgtca ggggggcgga gcctatggaa 4141 aaacgccagc aacgcagaaa ggcccacccg aaggtgagcc aggtgattac atttgggccc 4201 tcatcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc 4261 agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag 4321 tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt 4381 ttcctgtttg gtcacttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct 4441 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tgcgggaggg 4501 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga 4561 tttatcagca ataaaccagc cagccgggag ggccgagcgc agaagtgatc ctgcaacttt 4621 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt 4681 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt 4741 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat 4801 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgccagaa gtaagttggc 4861 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc 4921 cgtgagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat 4981 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag 5041 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgtaaactct caaggatctt 5101 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc 5161 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa 5221 gggaataagg gcgacacgga aatgttgaat actcatttta gcttccttag ctcctgaaaa 5281 tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt gaaagttgga 5341 acctcttacg tgccgatcaa gtcaaaagcc tccggtcgga ggcttttgac tttctgctat 5401 ggaggtcagg tatgatttaa atggtcagtt aga //