LOCUS Exported 6549 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Yeast Expression plasmid for SARS-CoV-2 M (membrane). ACCESSION . VERSION . KEYWORDS pGBW-m4046225 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6549) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 6549) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6549 /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 1144..1161 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" primer_bind complement(2304..2322) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 2673..3476 /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 3559..4541 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 4619..4648 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 4670..5257 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 5269..5296 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(5320..5511) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(5512..6372) /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 6135..6154 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6373..6475) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 6476..6507 /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 caactcgtgc cgagaggttc taaaatcgag gaaggtaagc tagtcatttg gataaatgga 1201 gataagggtt acaacggttt ggcagaagta ggcaagaaat ttgaaaagga cacaggtatc 1261 aaggtgacgg ttgaacatcc agacaagtta gaagaaaaat tccctcaagt tgctgctact 1321 ggtgatggtc ccgacattat tttctgggcc cacgatcgtt ttggtggata tgctcagtca 1381 ggtttgctgg ctgagatcac cccagacaag gccttccaag ataaattgta cccattcact 1441 tgggatgctg tcagatacaa cggtaagctt atagcatatc caattgctgt tgaagcctta 1501 tcgttgatct acaataagga cttgttacct aacccaccaa agacatggga agaaattcca 1561 gctttggata aagagctaaa ggccaaggga aaaagcgctt tgatgtttaa cttacaagaa 1621 ccttatttca cctggccatt aatcgctgct gacggtggtt acgcatttaa atacgaaaac 1681 ggtaagtatg acattaagga tgtcggcgtt gataatgctg gtgcaaaggc tggtttgacc 1741 ttcttggttg atttaataaa gaataaacac atgaacgctg acactgacta ctctatcgcc 1801 gaagccgctt tcaacaaggg tgaaactgct atgaccataa atggtccatg ggcctggagt 1861 aacattgata cttctaaagt aaactatggt gtcactgttt tgcctacatt caaaggccaa 1921 ccatccaagc cctttgtcgg tgttctatcc gctggcatca atgctgcttc tccaaacaag 1981 gagctagcta aagaattttt ggaaaattac ttgttaactg acgaaggttt ggaggccgtt 2041 aacaaggaca agccattggg tgctgtcgct ttgaaatctt acgaagaaga attggctaag 2101 gatcccagaa ttgccgctac catggagaac gcccaaaagg gtgaaattat gccaaacatc 2161 ccacaaatgt cagccttctg gtacgcagtt aggactgccg tgattaatgc tgcttccggt 2221 agacaaaccg tcgacgaagc attaaaagat gctcaaacgt aaacaggccc cttttccttt 2281 gtcgatatca tgtaattagt tatgtcacgc ttacattcac gccctcctcc cacatccgct 2341 ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta ttttttttaa 2401 tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt tttctgtaca 2461 aagcggccgc cgaaccacta gtgcactgca gtacacggtt tccttgaaat ttttttgatt 2521 cggtaatctc cgaacagaag gaagaacgaa ggaaggagca cagacttaga ttggtatata 2581 tacgcatatg tagtgttgaa gaaacatgaa attgcccagt attcttaacc caactgcaca 2641 gaacaaaaac gtgcaggaaa cgaagataaa tcatgtcgaa agctacatat aaggaacgtg 2701 ctgctactca tcctagtcct gttgctgcca agctatttaa tatcatgcac gaaaagcaaa 2761 caaacttgtg tgcttcattg gatgttcgta ccaccaagga attactggag ttagttgaag 2821 cattaggtcc caaaatttgt ttactaaaaa cacatgtgga tatcttgact gatttttcca 2881 tggagggcac agttaagccg ctaaaggcat tatccgccaa gtacaatttt ttactcttcg 2941 aggacagaaa atttgctgac attggtaata cagtcaaatt gcagtactct gcgggtgtat 3001 acagaatagc agaatgggca gacattacga atgcacacgg tgtggtgggc ccaggtattg 3061 ttagcggttt gaagcaggcg gcagaagaag taacaaagga acctagaggc cttttgatgt 3121 tagcagaatt gtcatgcaag ggctccctat ctactggaga atatactaag ggtactgttg 3181 acattgcgaa gagcgacaaa gattttgtta tcggctttat tgctcaaaga gacatgggtg 3241 gaagagatga aggttacgat tggttgatta tgacacccgg tgtgggttta gatgacaagg 3301 gagatgcatt gggtcaacag tatagaaccg tggatgatgt ggtttctaca ggatctgaca 3361 ttattattgt tggaagagga ctatttgcaa agggaaggga tgctaaggta gagggtgaac 3421 gttacagaaa agcaggctgg gaagcatatt tgagaagatg cggccagcaa aactaaaaaa 3481 ctgtattata agtaaatgca tgtatactaa actcacaaat tagagcttca atttaattat 3541 atcagttatt acccgagtaa cgaagcatct gtgcttcatt ttgtagaaca aaaatgcaac 3601 gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc 3661 aacgcgaaag cgctatttta ccaacgaaga atctgtgctt catttttgta aaacaaaaat 3721 gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga 3781 aatgcaacgc gagagcgcta ttttaccaac aaagaatcta tacttctttt ttgttctaca 3841 aaaatgcatc ccgagagcgc tatttttcta acaaagcatc ttagattact ttttttctcc 3901 tttgtgcgct ctataatgca gtctcttgat aactttttgc actgtaggtc cgttaaggtt 3961 agaagaaggc tactttggtg tctattttct cttccataaa aaaagcctga ctccacttcc 4021 cgcgtttact gattactagc gaagctgcgg gtgcattttt tcaagataaa ggcatccccg 4081 attatattct ataccgatgt ggattgcgca tactttgtga acagaaagtg atagcgttga 4141 tgattcttca ttggtcagaa aattatgaac ggtttcttct attttgtctc tatatactac 4201 gtataggaaa tgtttacatt ttcgtattgt tttcgattca ctctatgaat agttcttact 4261 acaatttttt tgtctaaaga gtaatactag agataaacat aaaaaatgta gaggtcgagt 4321 ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt tatataggga tatagcacag 4381 agatatatag caaagagata cttttgagca atgtttgtgg aagcggtatt cgcaatattt 4441 tagtagctcg ttacagtccg gtgcgttttt ggttttttga aagtgcgtca tcagagcgct 4501 tttggttttc aaaagcgctc tgaagttcct atactttcta gctagagaat aggaacttcc 4561 cgagcggccg cgtgttacaa gcttgactcc agcgtaactg gactgcaatc aactcactgg 4621 ctcaccttca cgggtgggcc tttcttcggt agaaaatcaa aggatcttct tgagatcctt 4681 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt 4741 gtttgccgga tcaagagcta ccaactcttt ttccgaggta actggcttca gcagagcgca 4801 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt 4861 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga 4921 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc 4981 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact 5041 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga 5101 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggt 5161 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcatcgatt 5221 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cagaaaggcc 5281 cacccgaagg tgagccaggt gattacattt gggccctcat cagaggtttt caccgtcatc 5341 accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca 5401 gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg 5461 gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca cttaccaatg 5521 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg 5581 actccccgtc gtgtagataa ctacgatgcg ggagggctta ccatctggcc ccagtgctgc 5641 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc 5701 cgggagggcc gagcgcagaa gtgatcctgc aactttatcc gcctccatcc agtctattaa 5761 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc 5821 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg 5881 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc 5941 cttcggtcct ccgatcgttg ccagaagtaa gttggccgca gtgttatcac tcatggttat 6001 ggcagcactg cataattctc ttactgtcat gccatccgtg agatgctttt ctgtgactgg 6061 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc 6121 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg 6181 aaaacgttct tcggggcgta aactctcaag gatcttaccg ctgttgagat ccagttcgat 6241 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg 6301 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg 6361 ttgaatactc attttagctt ccttagctcc tgaaaatctc gataactcaa aaaatacgcc 6421 cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc gatcaagtca 6481 aaagcctccg gtcggaggct tttgactttc tgctatggag gtcaggtatg atttaaatgg 6541 tcagttaga //