LOCUS Exported 6757 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Yeast Expression plasmid for SARS-CoV-2 3C-like proteinase. ACCESSION . VERSION . KEYWORDS pGBW-m4046557 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6757) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 6757) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6757 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 1352..1369 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" primer_bind complement(2512..2530) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 2881..3684 /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 3767..4749 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 4827..4856 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 4878..5465 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 5477..5504 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(5528..5719) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(5720..6580) /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 6343..6362 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6581..6683) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 6684..6715 /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 caataaaann nnnnntcacg 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 agaggttcta aaatcgagga 1381 aggtaagcta gtcatttgga taaatggaga taagggttac aacggtttgg cagaagtagg 1441 caagaaattt gaaaaggaca caggtatcaa ggtgacggtt gaacatccag acaagttaga 1501 agaaaaattc cctcaagttg ctgctactgg tgatggtccc gacattattt tctgggccca 1561 cgatcgtttt ggtggatatg ctcagtcagg tttgctggct gagatcaccc cagacaaggc 1621 cttccaagat aaattgtacc cattcacttg ggatgctgtc agatacaacg gtaagcttat 1681 agcatatcca attgctgttg aagccttatc gttgatctac aataaggact tgttacctaa 1741 cccaccaaag acatgggaag aaattccagc tttggataaa gagctaaagg ccaagggaaa 1801 aagcgctttg atgtttaact tacaagaacc ttatttcacc tggccattaa tcgctgctga 1861 cggtggttac gcatttaaat acgaaaacgg taagtatgac attaaggatg tcggcgttga 1921 taatgctggt gcaaaggctg gtttgacctt cttggttgat ttaataaaga ataaacacat 1981 gaacgctgac actgactact ctatcgccga agccgctttc aacaagggtg aaactgctat 2041 gaccataaat ggtccatggg cctggagtaa cattgatact tctaaagtaa actatggtgt 2101 cactgttttg cctacattca aaggccaacc atccaagccc tttgtcggtg ttctatccgc 2161 tggcatcaat gctgcttctc caaacaagga gctagctaaa gaatttttgg aaaattactt 2221 gttaactgac gaaggtttgg aggccgttaa caaggacaag ccattgggtg ctgtcgcttt 2281 gaaatcttac gaagaagaat tggctaagga tcccagaatt gccgctacca tggagaacgc 2341 ccaaaagggt gaaattatgc caaacatccc acaaatgtca gccttctggt acgcagttag 2401 gactgccgtg attaatgctg cttccggtag acaaaccgtc gacgaagcat taaaagatgc 2461 tcaaacgtaa acaggcccct tttcctttgt cgatatcatg taattagtta tgtcacgctt 2521 acattcacgc cctcctccca catccgctct aaccgaaaag gaaggagtta gacaacctga 2581 agtctaggtc cctatttatt ttttttaata gttatgttag tattaagaac gttatttata 2641 tttcaaattt ttcttttttt tctgtacaaa gcggccgccg aaccactagt gcactgcagt 2701 acacggtttc cttgaaattt ttttgattcg gtaatctccg aacagaagga agaacgaagg 2761 aaggagcaca gacttagatt ggtatatata cgcatatgta gtgttgaaga aacatgaaat 2821 tgcccagtat tcttaaccca actgcacaga acaaaaacgt gcaggaaacg aagataaatc 2881 atgtcgaaag ctacatataa ggaacgtgct gctactcatc ctagtcctgt tgctgccaag 2941 ctatttaata tcatgcacga aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc 3001 accaaggaat tactggagtt agttgaagca ttaggtccca aaatttgttt actaaaaaca 3061 catgtggata tcttgactga tttttccatg gagggcacag ttaagccgct aaaggcatta 3121 tccgccaagt acaatttttt actcttcgag gacagaaaat ttgctgacat tggtaataca 3181 gtcaaattgc agtactctgc gggtgtatac agaatagcag aatgggcaga cattacgaat 3241 gcacacggtg tggtgggccc aggtattgtt agcggtttga agcaggcggc agaagaagta 3301 acaaaggaac ctagaggcct tttgatgtta gcagaattgt catgcaaggg ctccctatct 3361 actggagaat atactaaggg tactgttgac attgcgaaga gcgacaaaga ttttgttatc 3421 ggctttattg ctcaaagaga catgggtgga agagatgaag gttacgattg gttgattatg 3481 acacccggtg tgggtttaga tgacaaggga gatgcattgg gtcaacagta tagaaccgtg 3541 gatgatgtgg tttctacagg atctgacatt attattgttg gaagaggact atttgcaaag 3601 ggaagggatg ctaaggtaga gggtgaacgt tacagaaaag caggctggga agcatatttg 3661 agaagatgcg gccagcaaaa ctaaaaaact gtattataag taaatgcatg tatactaaac 3721 tcacaaatta gagcttcaat ttaattatat cagttattac ccgagtaacg aagcatctgt 3781 gcttcatttt gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct 3841 gagctgcatt tttacagaac agaaatgcaa cgcgaaagcg ctattttacc aacgaagaat 3901 ctgtgcttca tttttgtaaa acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag 3961 aatctgagct gcatttttac agaacagaaa tgcaacgcga gagcgctatt ttaccaacaa 4021 agaatctata cttctttttt gttctacaaa aatgcatccc gagagcgcta tttttctaac 4081 aaagcatctt agattacttt ttttctcctt tgtgcgctct ataatgcagt ctcttgataa 4141 ctttttgcac tgtaggtccg ttaaggttag aagaaggcta ctttggtgtc tattttctct 4201 tccataaaaa aagcctgact ccacttcccg cgtttactga ttactagcga agctgcgggt 4261 gcattttttc aagataaagg catccccgat tatattctat accgatgtgg attgcgcata 4321 ctttgtgaac agaaagtgat agcgttgatg attcttcatt ggtcagaaaa ttatgaacgg 4381 tttcttctat tttgtctcta tatactacgt ataggaaatg tttacatttt cgtattgttt 4441 tcgattcact ctatgaatag ttcttactac aatttttttg tctaaagagt aatactagag 4501 ataaacataa aaaatgtaga ggtcgagttt agatgcaagt tcaaggagcg aaaggtggat 4561 gggtaggtta tatagggata tagcacagag atatatagca aagagatact tttgagcaat 4621 gtttgtggaa gcggtattcg caatatttta gtagctcgtt acagtccggt gcgtttttgg 4681 ttttttgaaa gtgcgtcatc agagcgcttt tggttttcaa aagcgctctg aagttcctat 4741 actttctagc tagagaatag gaacttcccg agcggccgcg tgttacaagc ttgactccag 4801 cgtaactgga ctgcaatcaa ctcactggct caccttcacg ggtgggcctt tcttcggtag 4861 aaaatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca 4921 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt 4981 ccgaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt 5041 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc 5101 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac 5161 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca 5221 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg 5281 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag 5341 gagagcgcac gagggagctt ccaggggtaa acgcctggta tctttatagt cctgtcgggt 5401 ttcgccacct ctgacttgag catcgatttt tgtgatgctc gtcagggggg cggagcctat 5461 ggaaaaacgc cagcaacgca gaaaggccca cccgaaggtg agccaggtga ttacatttgg 5521 gccctcatca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct 5581 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt 5641 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg 5701 ttttttcctg tttggtcact taccaatgct taatcagtga ggcacctatc tcagcgatct 5761 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatgcggg 5821 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 5881 cagatttatc agcaataaac cagccagccg ggagggccga gcgcagaagt gatcctgcaa 5941 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 6001 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt 6061 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 6121 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgcc agaagtaagt 6181 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 6241 catccgtgag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 6301 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata 6361 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgtaaa ctctcaagga 6421 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 6481 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 6541 aaaagggaat aagggcgaca cggaaatgtt gaatactcat tttagcttcc ttagctcctg 6601 aaaatctcga taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt 6661 tggaacctct tacgtgccga tcaagtcaaa agcctccggt cggaggcttt tgactttctg 6721 ctatggaggt caggtatgat ttaaatggtc agttaga //