LOCUS Exported 6423 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Yeast Expression plasmid for SARS-CoV-2 leader protein. ACCESSION . VERSION . KEYWORDS pGBW-m4046362 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6423) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 6423) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6423 /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 1018..1035 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" primer_bind complement(2178..2196) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 2547..3350 /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 3433..4415 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 4493..4522 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 4544..5131 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 5143..5170 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(5194..5385) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(5386..6246) /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 6009..6028 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6247..6349) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 6350..6381 /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 gaatctttgg tccctggatt taacgagaaa acccacgtac agctatccct tcccgtgttg 541 caagttcgtg acgttctggt tcggggtttc ggtgattcag tcgaagaagt cttgagtgaa 601 gccaggcaac atttaaagga tggtacttgt ggcttagttg aagtcgaaaa gggtgtgttg 661 ccacaacttg agcaaccata cgttttcatt aagagatctg acgcaagaac agcgccacac 721 ggtcatgtta tggttgaatt ggtcgctgaa ttggagggta tacaatatgg cagatcgggt 781 gaaactttag gtgtattggt cccacacgta ggtgaaatcc ctgttgctta cagaaaagtt 841 ttattaagaa agaatggtaa caagggtgct ggtggacact cctacggagc cgatctgaag 901 tcttttgact tgggtgatga attgggtact gatccatatg aagacttcca agaaaattgg 961 aacaccaaac atagctctgg tgttacgcgt gaattgatga gagaacttaa tggtggtctc 1021 gtgccgagag gttctaaaat cgaggaaggt aagctagtca tttggataaa tggagataag 1081 ggttacaacg gtttggcaga agtaggcaag aaatttgaaa aggacacagg tatcaaggtg 1141 acggttgaac atccagacaa gttagaagaa aaattccctc aagttgctgc tactggtgat 1201 ggtcccgaca ttattttctg ggcccacgat cgttttggtg gatatgctca gtcaggtttg 1261 ctggctgaga tcaccccaga caaggccttc caagataaat tgtacccatt cacttgggat 1321 gctgtcagat acaacggtaa gcttatagca tatccaattg ctgttgaagc cttatcgttg 1381 atctacaata aggacttgtt acctaaccca ccaaagacat gggaagaaat tccagctttg 1441 gataaagagc taaaggccaa gggaaaaagc gctttgatgt ttaacttaca agaaccttat 1501 ttcacctggc cattaatcgc tgctgacggt ggttacgcat ttaaatacga aaacggtaag 1561 tatgacatta aggatgtcgg cgttgataat gctggtgcaa aggctggttt gaccttcttg 1621 gttgatttaa taaagaataa acacatgaac gctgacactg actactctat cgccgaagcc 1681 gctttcaaca agggtgaaac tgctatgacc ataaatggtc catgggcctg gagtaacatt 1741 gatacttcta aagtaaacta tggtgtcact gttttgccta cattcaaagg ccaaccatcc 1801 aagccctttg tcggtgttct atccgctggc atcaatgctg cttctccaaa caaggagcta 1861 gctaaagaat ttttggaaaa ttacttgtta actgacgaag gtttggaggc cgttaacaag 1921 gacaagccat tgggtgctgt cgctttgaaa tcttacgaag aagaattggc taaggatccc 1981 agaattgccg ctaccatgga gaacgcccaa aagggtgaaa ttatgccaaa catcccacaa 2041 atgtcagcct tctggtacgc agttaggact gccgtgatta atgctgcttc cggtagacaa 2101 accgtcgacg aagcattaaa agatgctcaa acgtaaacag gccccttttc ctttgtcgat 2161 atcatgtaat tagttatgtc acgcttacat tcacgccctc ctcccacatc cgctctaacc 2221 gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt ttaatagtta 2281 tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg tacaaagcgg 2341 ccgccgaacc actagtgcac tgcagtacac ggtttccttg aaattttttt gattcggtaa 2401 tctccgaaca gaaggaagaa cgaaggaagg agcacagact tagattggta tatatacgca 2461 tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt aacccaactg cacagaacaa 2521 aaacgtgcag gaaacgaaga taaatcatgt cgaaagctac atataaggaa cgtgctgcta 2581 ctcatcctag tcctgttgct gccaagctat ttaatatcat gcacgaaaag caaacaaact 2641 tgtgtgcttc attggatgtt cgtaccacca aggaattact ggagttagtt gaagcattag 2701 gtcccaaaat ttgtttacta aaaacacatg tggatatctt gactgatttt tccatggagg 2761 gcacagttaa gccgctaaag gcattatccg ccaagtacaa ttttttactc ttcgaggaca 2821 gaaaatttgc tgacattggt aatacagtca aattgcagta ctctgcgggt gtatacagaa 2881 tagcagaatg ggcagacatt acgaatgcac acggtgtggt gggcccaggt attgttagcg 2941 gtttgaagca ggcggcagaa gaagtaacaa aggaacctag aggccttttg atgttagcag 3001 aattgtcatg caagggctcc ctatctactg gagaatatac taagggtact gttgacattg 3061 cgaagagcga caaagatttt gttatcggct ttattgctca aagagacatg ggtggaagag 3121 atgaaggtta cgattggttg attatgacac ccggtgtggg tttagatgac aagggagatg 3181 cattgggtca acagtataga accgtggatg atgtggtttc tacaggatct gacattatta 3241 ttgttggaag aggactattt gcaaagggaa gggatgctaa ggtagagggt gaacgttaca 3301 gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa aaaactgtat 3361 tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa ttatatcagt 3421 tattacccga gtaacgaagc atctgtgctt cattttgtag aacaaaaatg caacgcgaga 3481 gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg 3541 aaagcgctat tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa aaatgcaacg 3601 cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa cagaaatgca 3661 acgcgagagc gctattttac caacaaagaa tctatacttc ttttttgttc tacaaaaatg 3721 catcccgaga gcgctatttt tctaacaaag catcttagat tacttttttt ctcctttgtg 3781 cgctctataa tgcagtctct tgataacttt ttgcactgta ggtccgttaa ggttagaaga 3841 aggctacttt ggtgtctatt ttctcttcca taaaaaaagc ctgactccac ttcccgcgtt 3901 tactgattac tagcgaagct gcgggtgcat tttttcaaga taaaggcatc cccgattata 3961 ttctataccg atgtggattg cgcatacttt gtgaacagaa agtgatagcg ttgatgattc 4021 ttcattggtc agaaaattat gaacggtttc ttctattttg tctctatata ctacgtatag 4081 gaaatgttta cattttcgta ttgttttcga ttcactctat gaatagttct tactacaatt 4141 tttttgtcta aagagtaata ctagagataa acataaaaaa tgtagaggtc gagtttagat 4201 gcaagttcaa ggagcgaaag gtggatgggt aggttatata gggatatagc acagagatat 4261 atagcaaaga gatacttttg agcaatgttt gtggaagcgg tattcgcaat attttagtag 4321 ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc gtcatcagag cgcttttggt 4381 tttcaaaagc gctctgaagt tcctatactt tctagctaga gaataggaac ttcccgagcg 4441 gccgcgtgtt acaagcttga ctccagcgta actggactgc aatcaactca ctggctcacc 4501 ttcacgggtg ggcctttctt cggtagaaaa tcaaaggatc ttcttgagat cctttttttc 4561 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc 4621 cggatcaaga gctaccaact ctttttccga ggtaactggc ttcagcagag cgcagatacc 4681 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc 4741 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc 4801 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg 4861 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata 4921 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta 4981 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag gggtaaacgc 5041 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcatc gatttttgtg 5101 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcagaaa ggcccacccg 5161 aaggtgagcc aggtgattac atttgggccc tcatcagagg ttttcaccgt catcaccgaa 5221 acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc 5281 tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct 5341 gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcacttacc aatgcttaat 5401 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 5461 cgtcgtgtag ataactacga tgcgggaggg cttaccatct ggccccagtg ctgcaatgat 5521 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccgggag 5581 ggccgagcgc agaagtgatc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 5641 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 5701 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 5761 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 5821 tcctccgatc gttgccagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 5881 actgcataat tctcttactg tcatgccatc cgtgagatgc ttttctgtga ctggtgagta 5941 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 6001 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 6061 ttcttcgggg cgtaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 6121 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 6181 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 6241 actcatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag 6301 tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa gtcaaaagcc 6361 tccggtcgga ggcttttgac tttctgctat ggaggtcagg tatgatttaa atggtcagtt 6421 aga //