LOCUS Exported 5683 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Yeast Expression plasmid for SARS-CoV-2 3C-like proteinase. ACCESSION . VERSION . KEYWORDS pGBW-m4046516 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5683) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 5683) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5683 /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 1352..1369 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" CDS 1370..1393 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" primer_bind complement(1438..1456) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 1807..2610 /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 2693..3675 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 3753..3782 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 3804..4391 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 4403..4430 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(4454..4645) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(4646..5506) /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 5269..5288 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5507..5609) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 5610..5641 /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 atgtcgggtt tcagaaaaat ggcctttcct agtggaaagg tcgagggctg 481 tatggtgcaa gtaacatgcg gtacgaccac tttgaacggt ttatggcttg acgatgttgt 541 ttactgtcca cgtcatgtca tctgtacttc agaagatatg ttgaatccaa actatgaaga 601 cctactgatt cgcaagtcta accacaattt cttagttcag gctggtaacg ttcaattgag 661 agttataggt cacagcatgc aaaattgtgt cttgaagtta aaagttgata ccgcaaaccc 721 gaagactcca aagtacaaat ttgtcagaat ccaacccggt caaaccttct ccgtcttggc 781 ttgttataac gggtccccat ctggtgtata ccagtgcgct atgaggccaa acttcactat 841 taagggttct tttttgaatg gttcttgtgg ctccgttggt tttaatatcg actacgattg 901 cgtcagtttc tgttacatgc atcacatgga attgcctaca ggtgttcacg ccggaaccga 961 cctagaaggg aacttctatg gtccattcgt tgatcggcaa actgctcaag ctgcgggtac 1021 ggataccaca attactgtaa atgtgcttgc ctggttgtac gctgctgtta taaacggtga 1081 ccgatggttt ttaaacagat tcactaccac attgaatgac tttaatctgg tcgctatgaa 1141 atataactac gaaccactca ctcaagatca tgtcgatatt ttaggtcctt tgtctgcaca 1201 aactggcatc gctgttttgg acatgtgtgc gtcccttaag gaactactcc aaaacggtat 1261 gaacgggaga acgattttgg gttcagccct attggaggac gaatttaccc cattcgatgt 1321 ggttagacag tgctctggag taaccttcca actcgtgccg agaggttctt ggagccatcc 1381 gcagtttgaa aaataaacag gccccttttc ctttgtcgat atcatgtaat tagttatgtc 1441 acgcttacat tcacgccctc ctcccacatc cgctctaacc gaaaaggaag gagttagaca 1501 acctgaagtc taggtcccta tttatttttt ttaatagtta tgttagtatt aagaacgtta 1561 tttatatttc aaatttttct tttttttctg tacaaagcgg ccgccgaacc actagtgcac 1621 tgcagtacac ggtttccttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa 1681 cgaaggaagg agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca 1741 tgaaattgcc cagtattctt aacccaactg cacagaacaa aaacgtgcag gaaacgaaga 1801 taaatcatgt cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct 1861 gccaagctat ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt 1921 cgtaccacca aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta 1981 aaaacacatg tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag 2041 gcattatccg ccaagtacaa ttttttactc ttcgaggaca gaaaatttgc tgacattggt 2101 aatacagtca aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt 2161 acgaatgcac acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa 2221 gaagtaacaa aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc 2281 ctatctactg gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt 2341 gttatcggct ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg 2401 attatgacac ccggtgtggg tttagatgac aagggagatg cattgggtca acagtataga 2461 accgtggatg atgtggtttc tacaggatct gacattatta ttgttggaag aggactattt 2521 gcaaagggaa gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca 2581 tatttgagaa gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata 2641 ctaaactcac aaattagagc ttcaatttaa ttatatcagt tattacccga gtaacgaagc 2701 atctgtgctt cattttgtag aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa 2761 gaatctgagc tgcattttta cagaacagaa atgcaacgcg aaagcgctat tttaccaacg 2821 aagaatctgt gcttcatttt tgtaaaacaa aaatgcaacg cgagagcgct aatttttcaa 2881 acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgagagc gctattttac 2941 caacaaagaa tctatacttc ttttttgttc tacaaaaatg catcccgaga gcgctatttt 3001 tctaacaaag catcttagat tacttttttt ctcctttgtg cgctctataa tgcagtctct 3061 tgataacttt ttgcactgta ggtccgttaa ggttagaaga aggctacttt ggtgtctatt 3121 ttctcttcca taaaaaaagc ctgactccac ttcccgcgtt tactgattac tagcgaagct 3181 gcgggtgcat tttttcaaga taaaggcatc cccgattata ttctataccg atgtggattg 3241 cgcatacttt gtgaacagaa agtgatagcg ttgatgattc ttcattggtc agaaaattat 3301 gaacggtttc ttctattttg tctctatata ctacgtatag gaaatgttta cattttcgta 3361 ttgttttcga ttcactctat gaatagttct tactacaatt tttttgtcta aagagtaata 3421 ctagagataa acataaaaaa tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag 3481 gtggatgggt aggttatata gggatatagc acagagatat atagcaaaga gatacttttg 3541 agcaatgttt gtggaagcgg tattcgcaat attttagtag ctcgttacag tccggtgcgt 3601 ttttggtttt ttgaaagtgc gtcatcagag cgcttttggt tttcaaaagc gctctgaagt 3661 tcctatactt tctagctaga gaataggaac ttcccgagcg gccgcgtgtt acaagcttga 3721 ctccagcgta actggactgc aatcaactca ctggctcacc ttcacgggtg ggcctttctt 3781 cggtagaaaa tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg 3841 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact 3901 ctttttccga ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt 3961 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc 4021 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact 4081 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac 4141 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag 4201 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg 4261 gaacaggaga gcgcacgagg gagcttccag gggtaaacgc ctggtatctt tatagtcctg 4321 tcgggtttcg ccacctctga cttgagcatc gatttttgtg atgctcgtca ggggggcgga 4381 gcctatggaa aaacgccagc aacgcagaaa ggcccacccg aaggtgagcc aggtgattac 4441 atttgggccc tcatcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt 4501 aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca 4561 gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa 4621 gggcggtttt ttcctgtttg gtcacttacc aatgcttaat cagtgaggca cctatctcag 4681 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga 4741 tgcgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac 4801 cggctccaga tttatcagca ataaaccagc cagccgggag ggccgagcgc agaagtgatc 4861 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta 4921 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac 4981 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat 5041 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgccagaa 5101 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg 5161 tcatgccatc cgtgagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag 5221 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc 5281 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgtaaactct 5341 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat 5401 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg 5461 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatttta gcttccttag 5521 ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt 5581 gaaagttgga acctcttacg tgccgatcaa gtcaaaagcc tccggtcgga ggcttttgac 5641 tttctgctat ggaggtcagg tatgatttaa atggtcagtt aga //