LOCUS Exported 5386 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Yeast Expression plasmid for SARS-CoV-2 M (membrane). ACCESSION . VERSION . KEYWORDS pGBW-m4046626 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5386) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 5386) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5386 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 10..417 /gene="S. cerevisiae TEF1" /label=TEF1 promoter /note="promoter for EF-1-alpha" primer_bind complement(1141..1159) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 1510..2313 /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 2396..3378 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 3456..3485 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 3507..4094 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 4106..4133 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(4157..4348) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(4349..5209) /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 4972..4991 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5210..5312) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 5313..5344 /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 atggcagatt ccaacggtac tattaccgtt gaagagctta aaaagctcct 481 tgaacaatgg aacctagtaa taggtttcct attccttaca tggatttgtc ttctacaatt 541 tgcctatgcc aacaggaata ggtttttgta tataattaag ttaattttcc tctggctgtt 601 atggccagta actttagctt gttttgtgct tgctgctgtt tacagaataa attggatcac 661 cggtggaatt gctatcgcaa tggcttgtct tgtaggcttg atgtggctca gctacttcat 721 tgcttctttc agactgtttg cgcgtacgcg ttccatgtgg tcattcaatc cagaaactaa 781 cattcttctc aacgtgccac tccatggcac tattctgacc agaccgcttc tagaaagtga 841 actcgtaatc ggagctgtga tccttcgtgg acatcttcgt attgctggac accatctagg 901 acgctgtgac atcaaggacc tgcctaaaga aatcactgtt gctacatcac gaacgctttc 961 ttattacaaa ttgggagctt cgcagcgtgt agcaggtgac tcaggttttg ctgcatacag 1021 tcgctacagg attggcaact ataaattaaa cacagaccat tccagtagca gtgacaatat 1081 tgctttgctt gtacagtaaa caggcccctt ttcctttgtc gatatcatgt aattagttat 1141 gtcacgctta cattcacgcc ctcctcccac atccgctcta accgaaaagg aaggagttag 1201 acaacctgaa gtctaggtcc ctatttattt tttttaatag ttatgttagt attaagaacg 1261 ttatttatat ttcaaatttt tctttttttt ctgtacaaag cggccgccga accactagtg 1321 cactgcagta cacggtttcc ttgaaatttt tttgattcgg taatctccga acagaaggaa 1381 gaacgaagga aggagcacag acttagattg gtatatatac gcatatgtag tgttgaagaa 1441 acatgaaatt gcccagtatt cttaacccaa ctgcacagaa caaaaacgtg caggaaacga 1501 agataaatca tgtcgaaagc tacatataag gaacgtgctg ctactcatcc tagtcctgtt 1561 gctgccaagc tatttaatat catgcacgaa aagcaaacaa acttgtgtgc ttcattggat 1621 gttcgtacca ccaaggaatt actggagtta gttgaagcat taggtcccaa aatttgttta 1681 ctaaaaacac atgtggatat cttgactgat ttttccatgg agggcacagt taagccgcta 1741 aaggcattat ccgccaagta caatttttta ctcttcgagg acagaaaatt tgctgacatt 1801 ggtaatacag tcaaattgca gtactctgcg ggtgtataca gaatagcaga atgggcagac 1861 attacgaatg cacacggtgt ggtgggccca ggtattgtta gcggtttgaa gcaggcggca 1921 gaagaagtaa caaaggaacc tagaggcctt ttgatgttag cagaattgtc atgcaagggc 1981 tccctatcta ctggagaata tactaagggt actgttgaca ttgcgaagag cgacaaagat 2041 tttgttatcg gctttattgc tcaaagagac atgggtggaa gagatgaagg ttacgattgg 2101 ttgattatga cacccggtgt gggtttagat gacaagggag atgcattggg tcaacagtat 2161 agaaccgtgg atgatgtggt ttctacagga tctgacatta ttattgttgg aagaggacta 2221 tttgcaaagg gaagggatgc taaggtagag ggtgaacgtt acagaaaagc aggctgggaa 2281 gcatatttga gaagatgcgg ccagcaaaac taaaaaactg tattataagt aaatgcatgt 2341 atactaaact cacaaattag agcttcaatt taattatatc agttattacc cgagtaacga 2401 agcatctgtg cttcattttg tagaacaaaa atgcaacgcg agagcgctaa tttttcaaac 2461 aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgaaagcgc tattttacca 2521 acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca acgcgagagc gctaattttt 2581 caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgag agcgctattt 2641 taccaacaaa gaatctatac ttcttttttg ttctacaaaa atgcatcccg agagcgctat 2701 ttttctaaca aagcatctta gattactttt tttctccttt gtgcgctcta taatgcagtc 2761 tcttgataac tttttgcact gtaggtccgt taaggttaga agaaggctac tttggtgtct 2821 attttctctt ccataaaaaa agcctgactc cacttcccgc gtttactgat tactagcgaa 2881 gctgcgggtg cattttttca agataaaggc atccccgatt atattctata ccgatgtgga 2941 ttgcgcatac tttgtgaaca gaaagtgata gcgttgatga ttcttcattg gtcagaaaat 3001 tatgaacggt ttcttctatt ttgtctctat atactacgta taggaaatgt ttacattttc 3061 gtattgtttt cgattcactc tatgaatagt tcttactaca atttttttgt ctaaagagta 3121 atactagaga taaacataaa aaatgtagag gtcgagttta gatgcaagtt caaggagcga 3181 aaggtggatg ggtaggttat atagggatat agcacagaga tatatagcaa agagatactt 3241 ttgagcaatg tttgtggaag cggtattcgc aatattttag tagctcgtta cagtccggtg 3301 cgtttttggt tttttgaaag tgcgtcatca gagcgctttt ggttttcaaa agcgctctga 3361 agttcctata ctttctagct agagaatagg aacttcccga gcggccgcgt gttacaagct 3421 tgactccagc gtaactggac tgcaatcaac tcactggctc accttcacgg gtgggccttt 3481 cttcggtaga aaatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc 3541 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca 3601 actctttttc cgaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag 3661 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc 3721 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg 3781 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca 3841 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat 3901 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg 3961 tcggaacagg agagcgcacg agggagcttc caggggtaaa cgcctggtat ctttatagtc 4021 ctgtcgggtt tcgccacctc tgacttgagc atcgattttt gtgatgctcg tcaggggggc 4081 ggagcctatg gaaaaacgcc agcaacgcag aaaggcccac ccgaaggtga gccaggtgat 4141 tacatttggg ccctcatcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc 4201 ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt 4261 ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt 4321 taagggcggt tttttcctgt ttggtcactt accaatgctt aatcagtgag gcacctatct 4381 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta 4441 cgatgcggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct 4501 caccggctcc agatttatca gcaataaacc agccagccgg gagggccgag cgcagaagtg 4561 atcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa 4621 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt 4681 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta 4741 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgcca 4801 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta 4861 ctgtcatgcc atccgtgaga tgcttttctg tgactggtga gtactcaacc aagtcattct 4921 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg 4981 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgtaaac 5041 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact 5101 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa 5161 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcatt ttagcttcct 5221 tagctcctga aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat 5281 ggtgaaagtt ggaacctctt acgtgccgat caagtcaaaa gcctccggtc ggaggctttt 5341 gactttctgc tatggaggtc aggtatgatt taaatggtca gttaga //