LOCUS Exported 5635 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Yeast Expression plasmid for SARS-CoV-2 3C-like proteinase. ACCESSION . VERSION . KEYWORDS pGBW-m4046695 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5635) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 5635) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5635 /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(1390..1408) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 1759..2562 /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 2645..3627 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 3705..3734 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 3756..4343 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 4355..4382 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(4406..4597) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(4598..5458) /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 5221..5240 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5459..5561) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 5562..5593 /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 atgggtttta gaaaaatggc attcccatct ggtaaagttg agggttgtat 481 ggtacaagta acttgtggta caactacact taacggtctt tggcttgatg acgtagttta 541 ctgtccaaga catgtgatct gcacctctga agacatgctt aaccctaatt atgaagattt 601 actcattcgt aagtctaatc ataatttctt ggtacaggct ggtaatgttc aactcagggt 661 tattggacat tctatgcaaa attgtgtact taagcttaag gttgatacag ccaatcctaa 721 gacacctaag tataagtttg ttcgcattca accaggacag actttttcag tgttagcttg 781 ttacaatggt tcaccatctg gtgtttacca atgtgctatg aggcccaatt tcactattaa 841 gggttcattc cttaatggtt catgtggtag tgttggtttt aacatagatt atgactgtgt 901 ctctttttgt tacatgcacc atatggaatt accaactgga gttcatgctg gcacagactt 961 agaaggtaac ttttatggac cttttgttga caggcaaaca gcacaagcag ctggtacgga 1021 cacaactatt acagttaatg ttttagcttg gttgtacgct gctgttataa atggagacag 1081 gtggtttctc aatcgattta ccacaactct taatgacttt aaccttgtgg ctatgaagta 1141 caattatgaa cctctaacac aagaccatgt tgacatacta ggacctcttt ctgctcaaac 1201 tggaattgcc gttttagata tgtgtgcttc attaaaagaa ttactgcaaa atggtatgaa 1261 tggacgtacc atattgggta gtgctttatt agaagatgaa tttacacctt ttgatgttgt 1321 tagacaatgc tcaggtgtta ctttctaaac aggccccttt tcctttgtcg atatcatgta 1381 attagttatg tcacgcttac attcacgccc tcctcccaca tccgctctaa ccgaaaagga 1441 aggagttaga caacctgaag tctaggtccc tatttatttt ttttaatagt tatgttagta 1501 ttaagaacgt tatttatatt tcaaattttt cttttttttc tgtacaaagc ggccgccgaa 1561 ccactagtgc actgcagtac acggtttcct tgaaattttt ttgattcggt aatctccgaa 1621 cagaaggaag aacgaaggaa ggagcacaga cttagattgg tatatatacg catatgtagt 1681 gttgaagaaa catgaaattg cccagtattc ttaacccaac tgcacagaac aaaaacgtgc 1741 aggaaacgaa gataaatcat gtcgaaagct acatataagg aacgtgctgc tactcatcct 1801 agtcctgttg ctgccaagct atttaatatc atgcacgaaa agcaaacaaa cttgtgtgct 1861 tcattggatg ttcgtaccac caaggaatta ctggagttag ttgaagcatt aggtcccaaa 1921 atttgtttac taaaaacaca tgtggatatc ttgactgatt tttccatgga gggcacagtt 1981 aagccgctaa aggcattatc cgccaagtac aattttttac tcttcgagga cagaaaattt 2041 gctgacattg gtaatacagt caaattgcag tactctgcgg gtgtatacag aatagcagaa 2101 tgggcagaca ttacgaatgc acacggtgtg gtgggcccag gtattgttag cggtttgaag 2161 caggcggcag aagaagtaac aaaggaacct agaggccttt tgatgttagc agaattgtca 2221 tgcaagggct ccctatctac tggagaatat actaagggta ctgttgacat tgcgaagagc 2281 gacaaagatt ttgttatcgg ctttattgct caaagagaca tgggtggaag agatgaaggt 2341 tacgattggt tgattatgac acccggtgtg ggtttagatg acaagggaga tgcattgggt 2401 caacagtata gaaccgtgga tgatgtggtt tctacaggat ctgacattat tattgttgga 2461 agaggactat ttgcaaaggg aagggatgct aaggtagagg gtgaacgtta cagaaaagca 2521 ggctgggaag catatttgag aagatgcggc cagcaaaact aaaaaactgt attataagta 2581 aatgcatgta tactaaactc acaaattaga gcttcaattt aattatatca gttattaccc 2641 gagtaacgaa gcatctgtgc ttcattttgt agaacaaaaa tgcaacgcga gagcgctaat 2701 ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg cgaaagcgct 2761 attttaccaa cgaagaatct gtgcttcatt tttgtaaaac aaaaatgcaa cgcgagagcg 2821 ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg caacgcgaga 2881 gcgctatttt accaacaaag aatctatact tcttttttgt tctacaaaaa tgcatcccga 2941 gagcgctatt tttctaacaa agcatcttag attacttttt ttctcctttg tgcgctctat 3001 aatgcagtct cttgataact ttttgcactg taggtccgtt aaggttagaa gaaggctact 3061 ttggtgtcta ttttctcttc cataaaaaaa gcctgactcc acttcccgcg tttactgatt 3121 actagcgaag ctgcgggtgc attttttcaa gataaaggca tccccgatta tattctatac 3181 cgatgtggat tgcgcatact ttgtgaacag aaagtgatag cgttgatgat tcttcattgg 3241 tcagaaaatt atgaacggtt tcttctattt tgtctctata tactacgtat aggaaatgtt 3301 tacattttcg tattgttttc gattcactct atgaatagtt cttactacaa tttttttgtc 3361 taaagagtaa tactagagat aaacataaaa aatgtagagg tcgagtttag atgcaagttc 3421 aaggagcgaa aggtggatgg gtaggttata tagggatata gcacagagat atatagcaaa 3481 gagatacttt tgagcaatgt ttgtggaagc ggtattcgca atattttagt agctcgttac 3541 agtccggtgc gtttttggtt ttttgaaagt gcgtcatcag agcgcttttg gttttcaaaa 3601 gcgctctgaa gttcctatac tttctagcta gagaatagga acttcccgag cggccgcgtg 3661 ttacaagctt gactccagcg taactggact gcaatcaact cactggctca ccttcacggg 3721 tgggcctttc ttcggtagaa aatcaaagga tcttcttgag atcctttttt tctgcgcgta 3781 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa 3841 gagctaccaa ctctttttcc gaggtaactg gcttcagcag agcgcagata ccaaatactg 3901 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat 3961 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta 4021 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg 4081 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc 4141 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa 4201 gcggcagggt cggaacagga gagcgcacga gggagcttcc aggggtaaac gcctggtatc 4261 tttatagtcc tgtcgggttt cgccacctct gacttgagca tcgatttttg tgatgctcgt 4321 caggggggcg gagcctatgg aaaaacgcca gcaacgcaga aaggcccacc cgaaggtgag 4381 ccaggtgatt acatttgggc cctcatcaga ggttttcacc gtcatcaccg aaacgcgcga 4441 ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt 4501 catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc 4561 gggccatgtt aagggcggtt ttttcctgtt tggtcactta ccaatgctta atcagtgagg 4621 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt 4681 agataactac gatgcgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag 4741 acccacgctc accggctcca gatttatcag caataaacca gccagccggg agggccgagc 4801 gcagaagtga tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag 4861 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca 4921 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa 4981 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga 5041 tcgttgccag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata 5101 attctcttac tgtcatgcca tccgtgagat gcttttctgt gactggtgag tactcaacca 5161 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg 5221 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg 5281 ggcgtaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg 5341 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag 5401 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcattt 5461 tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta 5521 tttcattatg gtgaaagttg gaacctctta cgtgccgatc aagtcaaaag cctccggtcg 5581 gaggcttttg actttctgct atggaggtca ggtatgattt aaatggtcag ttaga //