LOCUS Exported 8575 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Yeast Expression plasmid for SARS-CoV-2 surface glycoprotein (Spike protein). ACCESSION . VERSION . KEYWORDS pGBW-m4046494 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8575) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 8575) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8575 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 18..425 /gene="S. cerevisiae TEF1" /label=TEF1 promoter /note="promoter for EF-1-alpha" CDS 4258..4275 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" CDS 4276..4293 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(4338..4356) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 4707..5510 /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 5593..6575 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 6653..6682 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 6704..7291 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 7303..7330 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(7354..7545) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(7546..8406) /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 8169..8188 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8407..8509) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 8510..8541 /label=tonB terminator /note="bidirectional E. coli tonB-P14 transcription terminator" ORIGIN 1 cagttagagc ggccgcccca cacaccatag cttcaaaatg tttctactcc ttttttactc 61 ttccagattt tctcggactc cgcgcatcgc cgtaccactt caaaacaccc aagcacagca 121 tactaaattt cccctctttc ttcctctagg gtgtcgttaa ttacccgtac taaaggtttg 181 gaaaagaaaa aagagaccgc ctcgtttctt tttcttcgtc gaaaaaggca ataaaaattt 241 ttatcacgtt tctttttctt gaaaattttt ttttttgatt tttttctctt tcgatgacct 301 cccattgata tttaagttaa taaacggtct tcaatttctc aagtttcagt ttcatttttc 361 ttgttctatt acaacttttt ttacttcttg ctcattagaa agaaagcata gcaatctaat 421 ctaagtttta attacaaaat gttcgtcttt ttagttttgc ttccattggt gagctcccaa 481 tgcgttaatc tgacaactcg cacgcagcta cctccagcat acactaactc tttcaccaga 541 ggcgtctatt accccgacaa agtatttcgt tcttcggttt tacatagtac ccaagatttg 601 ttcttgccat tcttttcaaa cgttacttgg tttcacgcca tacacgtttc tggtaccaat 661 ggtactaaga ggttcgataa cccagtgttg ccattcaatg acggtgtcta cttcgcttcc 721 acagaaaagt ctaacatcat tagaggttgg atctttggaa ccacgcttga ttccaagact 781 caatctttgt taattgtcaa caacgctact aatgttgtaa ttaaagtctg tgagttccaa 841 ttctgtaacg acccgttttt gggtgtttat taccataaga ataataaaag ctggatggaa 901 tccgaatttc gggtttattc aagtgctaac aactgtacct tcgaatacgt cagtcaacct 961 ttcctgatgg atttggaggg taagcaaggc aacttcaaga acttgagaga atttgtcttc 1021 aaaaatatcg acggttactt taagatctac tctaagcaca caccaattaa cctagttaga 1081 gatttacctc agggtttctc tgctttggaa ccactcgttg atttgccgat tggtatcaac 1141 attacgagat ttcaaacttt gctagcctta caccgatcat atttaactcc tggtgactcc 1201 tcgtctggat ggactgcggg tgctgctgcc tactatgtcg gttatttgca gccaagaacc 1261 tttctgctca agtacaatga aaacgggaca attaccgatg ctgtagactg tgctttggat 1321 ccattgagcg aaactaagtg caccctaaaa tctttcacag ttgaaaaggg tatataccaa 1381 actagcaact ttagagttca accaactgaa tcaatagtca gattcccaaa tatcaccaac 1441 ttgtgtccat tcggtgaggt ttttaatgcc acaaggtttg cttctgtgta cgcttggaac 1501 cgtaagagaa tctccaattg tgtggcagat tactctgtac tatataacag tgcttccttc 1561 tctactttca aatgttacgg ggtttcgcca accaagctta acgacttatg cttcactaat 1621 gtttacgcag actcctttgt cattagaggc gatgaagtaa gacaaatcgc gccaggtcaa 1681 accggtaaga ttgctgatta caactacaaa ttgccagacg acttcaccgg ttgcgtgatc 1741 gcttggaata gcaacaactt ggattcgaag gttggtggga actataacta tttgtaccgc 1801 ctatttcgta agtctaattt gaaacctttc gaaagagaca taagtaccga gatttatcaa 1861 gctggttcta ctccttgtaa tggggtcgaa ggttttaact gttacttccc cttacaatct 1921 tacggttttc agccaaccaa cggagttggt taccaaccat atagagttgt tgtcctgtct 1981 tttgaactat tgcatgcccc agctacagtc tgcggtccca aaaagtccac gaacttggtt 2041 aaaaataagt gtgtcaactt caatttcaat ggtcttactg gtactggcgt attgaccgaa 2101 tccaacaaga agttcctgcc attccaacaa ttcggtagag atattgcaga tacgaccgat 2161 gcggtccggg atcctcaaac tttggaaatt ttggacatca ctccatgttc ctttggtggt 2221 gttagcgtta ttacccctgg aaccaacact tctaatcagg tcgccgtttt ataccaagac 2281 gtcaactgta cggaagtgcc agttgctatc cacgctgacc aactaacccc tacttggaga 2341 gtctactcta ctggttctaa tgttttccaa acaagagctg gttgtttgat aggtgctgag 2401 catgtcaaca actcctatga atgtgacatt ccaattggtg ccggaatctg tgcatcctac 2461 caaacacaga ctaattcacc aagaagagct agaagtgttg cttcacaaag tatcattgca 2521 tacaccatga gcctaggtgc cgaaaactca gtcgcataca gtaacaattc tattgctata 2581 ccaactaatt tcactatttc tgttactact gaaatcttac ctgtctctat gacaaagacc 2641 tcagtggatt gtacaatgta tatctgtggt gactctactg agtgttctaa tttgttgtta 2701 caatacggtt ccttctgcac acaattgaac cgtgctctta cgggtattgc tgtcgaacaa 2761 gacaaaaaca cccaggaagt atttgcccaa gtcaagcaaa tctacaagac cccacccatt 2821 aaggatttcg gcggtttcaa ctttagccaa atcttgccag atccatcaaa gccatccaag 2881 agatcgttta tcgaagattt attatttaat aaagttacgt tagccgacgc tggattcatc 2941 aagcaatatg gcgattgctt gggtgacatt gcagcccgag acttgatatg tgcgcagaag 3001 ttcaatggtc ttaccgtgtt gcccccacta ttgactgatg aaatgatcgc tcaatatacc 3061 tcagcgctct tggctgggac gattacttct ggttggacat ttggtgctgg tgctgcattg 3121 caaatcccat ttgctatgca aatggcttac agattcaacg ggataggagt tactcaaaac 3181 gttctgtatg aaaaccaaaa gctaatcgct aatcaattca actcggccat tggtaagatt 3241 caagactctt tgtcctcgac tgcttcggcg cttggtaaac tacaagacgt tgttaaccag 3301 aatgctcaag ctttgaacac cctcgtaaaa cagttgtcct ctaacttcgg ggccatttct 3361 tccgtactca acgatatctt atctaggttg gataaggttg aggccgaagt acaaatcgat 3421 agattgatca ccggaagatt gcaatcgcta caaacctacg ttactcaaca acttatccgc 3481 gctgctgaaa ttagagctag tgctaattta gccgcgacca agatgagtga atgtgtccta 3541 ggtcagtcta agcgtgttga cttctgtggt aaaggttatc atttgatgtc ctttccccaa 3601 tctgccccgc acggtgtcgt ctttttgcat gttacttacg tgccggctca ggaaaagaac 3661 ttcactacag ctcctgccat ctgtcacgat ggtaaagctc acttcccaag agaaggtgtt 3721 ttcgtttcta atggtactca ctggttcgtc acacaaagaa acttctacga gccccaaatt 3781 attactactg acaacacctt cgtatccggt aattgtgacg ttgtcatcgg tattgtcaac 3841 aacaccgttt atgatccact ccaaccagaa ttagattcct tcaaggaaga attggataaa 3901 tactttaaga atcacacctc ccctgacgtt gatctaggtg acatatctgg cattaacgct 3961 tccgtggtta acattcaaaa ggaaatagac aggttaaatg aggttgccaa gaacttgaac 4021 gagtcattga ttgatctcca agaactaggt aagtacgaac aatacattaa atggccgtgg 4081 tacatctggt tgggttttat cgcagggctg attgctattg ttatggtgac catcatgttg 4141 tgctgtatga cctcatgttg ttcttgttta aagggttgtt gcagctgcgg ttcttgttgc 4201 aagttcgatg aagacgactc tgaacctgta ttgaagggtg ttaaattgca ctacacgctc 4261 gtgccgagag gttctcacca tcatcaccat cattaaacag gccccttttc ctttgtcgat 4321 atcatgtaat tagttatgtc acgcttacat tcacgccctc ctcccacatc cgctctaacc 4381 gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt ttaatagtta 4441 tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg tacaaagcgg 4501 ccgccgaacc actagtgcac tgcagtacac ggtttccttg aaattttttt gattcggtaa 4561 tctccgaaca gaaggaagaa cgaaggaagg agcacagact tagattggta tatatacgca 4621 tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt aacccaactg cacagaacaa 4681 aaacgtgcag gaaacgaaga taaatcatgt cgaaagctac atataaggaa cgtgctgcta 4741 ctcatcctag tcctgttgct gccaagctat ttaatatcat gcacgaaaag caaacaaact 4801 tgtgtgcttc attggatgtt cgtaccacca aggaattact ggagttagtt gaagcattag 4861 gtcccaaaat ttgtttacta aaaacacatg tggatatctt gactgatttt tccatggagg 4921 gcacagttaa gccgctaaag gcattatccg ccaagtacaa ttttttactc ttcgaggaca 4981 gaaaatttgc tgacattggt aatacagtca aattgcagta ctctgcgggt gtatacagaa 5041 tagcagaatg ggcagacatt acgaatgcac acggtgtggt gggcccaggt attgttagcg 5101 gtttgaagca ggcggcagaa gaagtaacaa aggaacctag aggccttttg atgttagcag 5161 aattgtcatg caagggctcc ctatctactg gagaatatac taagggtact gttgacattg 5221 cgaagagcga caaagatttt gttatcggct ttattgctca aagagacatg ggtggaagag 5281 atgaaggtta cgattggttg attatgacac ccggtgtggg tttagatgac aagggagatg 5341 cattgggtca acagtataga accgtggatg atgtggtttc tacaggatct gacattatta 5401 ttgttggaag aggactattt gcaaagggaa gggatgctaa ggtagagggt gaacgttaca 5461 gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa aaaactgtat 5521 tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa ttatatcagt 5581 tattacccga gtaacgaagc atctgtgctt cattttgtag aacaaaaatg caacgcgaga 5641 gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg 5701 aaagcgctat tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa aaatgcaacg 5761 cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa cagaaatgca 5821 acgcgagagc gctattttac caacaaagaa tctatacttc ttttttgttc tacaaaaatg 5881 catcccgaga gcgctatttt tctaacaaag catcttagat tacttttttt ctcctttgtg 5941 cgctctataa tgcagtctct tgataacttt ttgcactgta ggtccgttaa ggttagaaga 6001 aggctacttt ggtgtctatt ttctcttcca taaaaaaagc ctgactccac ttcccgcgtt 6061 tactgattac tagcgaagct gcgggtgcat tttttcaaga taaaggcatc cccgattata 6121 ttctataccg atgtggattg cgcatacttt gtgaacagaa agtgatagcg ttgatgattc 6181 ttcattggtc agaaaattat gaacggtttc ttctattttg tctctatata ctacgtatag 6241 gaaatgttta cattttcgta ttgttttcga ttcactctat gaatagttct tactacaatt 6301 tttttgtcta aagagtaata ctagagataa acataaaaaa tgtagaggtc gagtttagat 6361 gcaagttcaa ggagcgaaag gtggatgggt aggttatata gggatatagc acagagatat 6421 atagcaaaga gatacttttg agcaatgttt gtggaagcgg tattcgcaat attttagtag 6481 ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc gtcatcagag cgcttttggt 6541 tttcaaaagc gctctgaagt tcctatactt tctagctaga gaataggaac ttcccgagcg 6601 gccgcgtgtt acaagcttga ctccagcgta actggactgc aatcaactca ctggctcacc 6661 ttcacgggtg ggcctttctt cggtagaaaa tcaaaggatc ttcttgagat cctttttttc 6721 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc 6781 cggatcaaga gctaccaact ctttttccga ggtaactggc ttcagcagag cgcagatacc 6841 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc 6901 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc 6961 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg 7021 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata 7081 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta 7141 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag gggtaaacgc 7201 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcatc gatttttgtg 7261 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcagaaa ggcccacccg 7321 aaggtgagcc aggtgattac atttgggccc tcatcagagg ttttcaccgt catcaccgaa 7381 acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc 7441 tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct 7501 gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcacttacc aatgcttaat 7561 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 7621 cgtcgtgtag ataactacga tgcgggaggg cttaccatct ggccccagtg ctgcaatgat 7681 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccgggag 7741 ggccgagcgc agaagtgatc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 7801 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 7861 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 7921 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 7981 tcctccgatc gttgccagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 8041 actgcataat tctcttactg tcatgccatc cgtgagatgc ttttctgtga ctggtgagta 8101 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 8161 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 8221 ttcttcgggg cgtaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 8281 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 8341 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 8401 actcatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag 8461 tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa gtcaaaagcc 8521 tccggtcgga ggcttttgac tttctgctat ggaggtcagg tatgatttaa atggt //