LOCUS Exported 5422 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Yeast Expression plasmid for SARS-CoV-2 M (membrane). ACCESSION . VERSION . KEYWORDS pGBW-m4046268 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5422) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 5422) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5422 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 10..417 /gene="S. cerevisiae TEF1" /label=TEF1 promoter /note="promoter for EF-1-alpha" CDS 1097..1114 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" CDS 1115..1132 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(1177..1195) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 1546..2349 /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 2432..3414 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 3492..3521 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 3543..4130 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 4142..4169 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(4193..4384) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(4385..5245) /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 5008..5027 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5246..5348) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 5349..5380 /label=tonB terminator /note="bidirectional E. coli tonB-P14 transcription terminator" ORIGIN 1 gcggccgccc cacacaccat agcttcaaaa tgtttctact ccttttttac tcttccagat 61 tttctcggac tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag catactaaat 121 ttcccctctt tcttcctcta gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa 181 aaaagagacc gcctcgtttc tttttcttcg tcgaaaaagg caataaaaat ttttatcacg 241 tttctttttc ttgaaaattt ttttttttga tttttttctc tttcgatgac ctcccattga 301 tatttaagtt aataaacggt cttcaatttc tcaagtttca gtttcatttt tcttgttcta 361 ttacaacttt ttttacttct tgctcattag aaagaaagca tagcaatcta atctaagttt 421 taattacaaa atggctgatt ccaatggcac catcacagta gaagagctga aaaagctcct 481 tgaacaatgg aacttggtta taggtttttt attcttgact tggatttgcc tattgcagtt 541 cgcctatgca aacagaaacc gttttttgta cattatcaag ttaattttct tgtggctgtt 601 atggcctgtc actcttgctt gttttgtttt ggctgccgtc taccggataa attggatcac 661 cggtggtatc gctattgcaa tggcttgttt ggttggattg atgtggttga gttatttcat 721 tgcctcattc agattattcg ctaggactag atcgatgtgg tcctttaatc cagaaacgaa 781 catcttgttg aacgtgccac tccatggtac tattttgacc agaccgctat tagaatctga 841 gttggttatc ggtgctgtca ttctaagagg gcacttacgt atcgcgggtc accatttggg 901 aagatgtgac attaaggatc ttccaaaaga aatcacagtc gctacctctc gaactctatc 961 ttactacaag ttgggtgcct cacaaagggt tgctggtgac agcggtttcg ctgcctattc 1021 tcgctaccgt atcggcaact acaaattaaa taccgatcac tcctctagtt ctgacaatat 1081 tgccttattg gtgcaactcg tgccgagagg ttctcaccat catcaccatc attaaacagg 1141 ccccttttcc tttgtcgata tcatgtaatt agttatgtca cgcttacatt cacgccctcc 1201 tcccacatcc gctctaaccg aaaaggaagg agttagacaa cctgaagtct aggtccctat 1261 ttattttttt taatagttat gttagtatta agaacgttat ttatatttca aatttttctt 1321 ttttttctgt acaaagcggc cgccgaacca ctagtgcact gcagtacacg gtttccttga 1381 aatttttttg attcggtaat ctccgaacag aaggaagaac gaaggaagga gcacagactt 1441 agattggtat atatacgcat atgtagtgtt gaagaaacat gaaattgccc agtattctta 1501 acccaactgc acagaacaaa aacgtgcagg aaacgaagat aaatcatgtc gaaagctaca 1561 tataaggaac gtgctgctac tcatcctagt cctgttgctg ccaagctatt taatatcatg 1621 cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc gtaccaccaa ggaattactg 1681 gagttagttg aagcattagg tcccaaaatt tgtttactaa aaacacatgt ggatatcttg 1741 actgattttt ccatggaggg cacagttaag ccgctaaagg cattatccgc caagtacaat 1801 tttttactct tcgaggacag aaaatttgct gacattggta atacagtcaa attgcagtac 1861 tctgcgggtg tatacagaat agcagaatgg gcagacatta cgaatgcaca cggtgtggtg 1921 ggcccaggta ttgttagcgg tttgaagcag gcggcagaag aagtaacaaa ggaacctaga 1981 ggccttttga tgttagcaga attgtcatgc aagggctccc tatctactgg agaatatact 2041 aagggtactg ttgacattgc gaagagcgac aaagattttg ttatcggctt tattgctcaa 2101 agagacatgg gtggaagaga tgaaggttac gattggttga ttatgacacc cggtgtgggt 2161 ttagatgaca agggagatgc attgggtcaa cagtatagaa ccgtggatga tgtggtttct 2221 acaggatctg acattattat tgttggaaga ggactatttg caaagggaag ggatgctaag 2281 gtagagggtg aacgttacag aaaagcaggc tgggaagcat atttgagaag atgcggccag 2341 caaaactaaa aaactgtatt ataagtaaat gcatgtatac taaactcaca aattagagct 2401 tcaatttaat tatatcagtt attacccgag taacgaagca tctgtgcttc attttgtaga 2461 acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac 2521 agaacagaaa tgcaacgcga aagcgctatt ttaccaacga agaatctgtg cttcattttt 2581 gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt 2641 tttacagaac agaaatgcaa cgcgagagcg ctattttacc aacaaagaat ctatacttct 2701 tttttgttct acaaaaatgc atcccgagag cgctattttt ctaacaaagc atcttagatt 2761 actttttttc tcctttgtgc gctctataat gcagtctctt gataactttt tgcactgtag 2821 gtccgttaag gttagaagaa ggctactttg gtgtctattt tctcttccat aaaaaaagcc 2881 tgactccact tcccgcgttt actgattact agcgaagctg cgggtgcatt ttttcaagat 2941 aaaggcatcc ccgattatat tctataccga tgtggattgc gcatactttg tgaacagaaa 3001 gtgatagcgt tgatgattct tcattggtca gaaaattatg aacggtttct tctattttgt 3061 ctctatatac tacgtatagg aaatgtttac attttcgtat tgttttcgat tcactctatg 3121 aatagttctt actacaattt ttttgtctaa agagtaatac tagagataaa cataaaaaat 3181 gtagaggtcg agtttagatg caagttcaag gagcgaaagg tggatgggta ggttatatag 3241 ggatatagca cagagatata tagcaaagag atacttttga gcaatgtttg tggaagcggt 3301 attcgcaata ttttagtagc tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg 3361 tcatcagagc gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt ctagctagag 3421 aataggaact tcccgagcgg ccgcgtgtta caagcttgac tccagcgtaa ctggactgca 3481 atcaactcac tggctcacct tcacgggtgg gcctttcttc ggtagaaaat caaaggatct 3541 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta 3601 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgag gtaactggct 3661 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact 3721 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg 3781 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata 3841 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga 3901 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag 3961 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg 4021 agcttccagg ggtaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac 4081 ttgagcatcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca 4141 acgcagaaag gcccacccga aggtgagcca ggtgattaca tttgggccct catcagaggt 4201 tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt 4261 gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa 4321 gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg 4381 tcacttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat 4441 ccatagttgc ctgactcccc gtcgtgtaga taactacgat gcgggagggc ttaccatctg 4501 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa 4561 taaaccagcc agccgggagg gccgagcgca gaagtgatcc tgcaacttta tccgcctcca 4621 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc 4681 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt 4741 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa 4801 aagcggttag ctccttcggt cctccgatcg ttgccagaag taagttggcc gcagtgttat 4861 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtgagatgct 4921 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga 4981 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag 5041 tgctcatcat tggaaaacgt tcttcggggc gtaaactctc aaggatctta ccgctgttga 5101 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca 5161 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg 5221 cgacacggaa atgttgaata ctcattttag cttccttagc tcctgaaaat ctcgataact 5281 caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt 5341 gccgatcaag tcaaaagcct ccggtcggag gcttttgact ttctgctatg gaggtcaggt 5401 atgatttaaa tggtcagtta ga //