LOCUS Exported 6733 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Yeast expression plasmid for SARS-CoV-2 2'-O-ribose methyltransferase. ACCESSION . VERSION . KEYWORDS pGBW-m4046279 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6733) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 6733) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6733 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(299..1102) /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" primer_bind 1453..1471 /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS complement(2614..2631) /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" promoter complement(3542..3949) /gene="S. cerevisiae TEF1" /label=TEF1 promoter /note="promoter for EF-1-alpha" terminator 4001..4032 /label=tonB terminator /note="bidirectional E. coli tonB-P14 transcription terminator" promoter 4033..4135 /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" CDS 4136..4996 /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 complement(4354..4373) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" CDS 4997..5188 /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" terminator 5212..5239 /label=T7Te terminator /note="phage T7 early transcription terminator" rep_origin complement(5251..5838) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 5860..5889 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 5967..216 /label=2u ori /note="yeast 2u plasmid origin of replication" ORIGIN 1 ctgtaaaaat gcagctcaga ttctttgttt gaaaaattag cgctctcgcg ttgcattttt 61 gttttacaaa aatgaagcac agattcttcg ttggtaaaat agcgctttcg cgttgcattt 121 ctgttctgta aaaatgcagc tcagattctt tgtttgaaaa attagcgctc tcgcgttgca 181 tttttgttct acaaaatgaa gcacagatgc ttcgttactc gggtaataac tgatataatt 241 aaattgaagc tctaatttgt gagtttagta tacatgcatt tacttataat acagtttttt 301 agttttgctg gccgcatctt ctcaaatatg cttcccagcc tgcttttctg taacgttcac 361 cctctacctt agcatccctt ccctttgcaa atagtcctct tccaacaata ataatgtcag 421 atcctgtaga aaccacatca tccacggttc tatactgttg acccaatgca tctcccttgt 481 catctaaacc cacaccgggt gtcataatca accaatcgta accttcatct cttccaccca 541 tgtctctttg agcaataaag ccgataacaa aatctttgtc gctcttcgca atgtcaacag 601 tacccttagt atattctcca gtagataggg agcccttgca tgacaattct gctaacatca 661 aaaggcctct aggttccttt gttacttctt ctgccgcctg cttcaaaccg ctaacaatac 721 ctgggcccac cacaccgtgt gcattcgtaa tgtctgccca ttctgctatt ctgtatacac 781 ccgcagagta ctgcaatttg actgtattac caatgtcagc aaattttctg tcctcgaaga 841 gtaaaaaatt gtacttggcg gataatgcct ttagcggctt aactgtgccc tccatggaaa 901 aatcagtcaa gatatccaca tgtgttttta gtaaacaaat tttgggacct aatgcttcaa 961 ctaactccag taattccttg gtggtacgaa catccaatga agcacacaag tttgtttgct 1021 tttcgtgcat gatattaaat agcttggcag caacaggact aggatgagta gcagcacgtt 1081 ccttatatgt agctttcgac atgatttatc ttcgtttcct gcacgttttt gttctgtgca 1141 gttgggttaa gaatactggg caatttcatg tttcttcaac actacatatg cgtatatata 1201 ccaatctaag tctgtgctcc ttccttcgtt cttccttctg ttcggagatt accgaatcaa 1261 aaaaatttca aggaaaccgt gtactgcagt gcactagtgg ttcggcggcc gctttgtaca 1321 gaaaaaaaag aaaaatttga aatataaata acgttcttaa tactaacata actattaaaa 1381 aaaataaata gggacctaga cttcaggttg tctaactcct tccttttcgg ttagagcgga 1441 tgtgggagga gggcgtgaat gtaagcgtga cataactaat tacatgatat cgacaaagga 1501 aaaggggcct gtttacgttt gagcatcttt taatgcttcg tcgacggttt gtctaccgga 1561 agcagcatta atcacggcag tcctaactgc gtaccagaag gctgacattt gtgggatgtt 1621 tggcataatt tcaccctttt gggcgttctc catggtagcg gcaattctgg gatccttagc 1681 caattcttct tcgtaagatt tcaaagcgac agcacccaat ggcttgtcct tgttaacggc 1741 ctccaaacct tcgtcagtta acaagtaatt ttccaaaaat tctttagcta gctccttgtt 1801 tggagaagca gcattgatgc cagcggatag aacaccgaca aagggcttgg atggttggcc 1861 tttgaatgta ggcaaaacag tgacaccata gtttacttta gaagtatcaa tgttactcca 1921 ggcccatgga ccatttatgg tcatagcagt ttcacccttg ttgaaagcgg cttcggcgat 1981 agagtagtca gtgtcagcgt tcatgtgttt attctttatt aaatcaacca agaaggtcaa 2041 accagccttt gcaccagcat tatcaacgcc gacatcctta atgtcatact taccgttttc 2101 gtatttaaat gcgtaaccac cgtcagcagc gattaatggc caggtgaaat aaggttcttg 2161 taagttaaac atcaaagcgc tttttccctt ggcctttagc tctttatcca aagctggaat 2221 ttcttcccat gtctttggtg ggttaggtaa caagtcctta ttgtagatca acgataaggc 2281 ttcaacagca attggatatg ctataagctt accgttgtat ctgacagcat cccaagtgaa 2341 tgggtacaat ttatcttgga aggccttgtc tggggtgatc tcagccagca aacctgactg 2401 agcatatcca ccaaaacgat cgtgggccca gaaaataatg tcgggaccat caccagtagc 2461 agcaacttga gggaattttt cttctaactt gtctggatgt tcaaccgtca ccttgatacc 2521 tgtgtccttt tcaaatttct tgcctacttc tgccaaaccg ttgtaaccct tatctccatt 2581 tatccaaatg actagcttac cttcctcgat tttagaacct ctcggcacga ggttgttgac 2641 tagcacatca gaagagataa caacccgatt attttctcta ataatgagtc tacctttaga 2701 cagcaaagat aagatcatat cgttgatctg cccttccttc aaggacatga cagctgtacc 2761 tctcaacttc aatggaaact ttgacatgtc gaataaacta tagcttgaca attgtatagg 2821 gttagtattt ctccagaaaa tgtagttagc gtgcatgacg taaccatcaa tttgttcgcg 2881 tggctttcct aaataattac aaccgatgag aaaggcttcc gaagaactag cgttaacgtt 2941 ggtaacaaac gcggtccacc aagcgaaatg acccatcaat ttgtataagt cggcgttcca 3001 ggagtgttca gtgatcttaa tagccacaga accgcccaag gcaagctttt gctggatgaa 3061 accacagata taagtaaaga aaccttcttt agaatcattt tcctttgtaa catttttggt 3121 ctttgggtcg tacatatccg aaataatcaa gtcccacttg ttcgccgtat gaacggtagc 3181 acaatcaccg attaaagtgg agtcagcatc actgacgaag tcgttcagat cagaatcaac 3241 caatagagta ccagttggga gccattgcct tagaactgcg gtacctggag cgacgccttt 3301 gtctgaaccg gccccaaagt gtatgacacg catgttatac ggaacagcca aagtcaacgt 3361 atttaagtat tgacacaatt gggtgtactt tgctacgttc atcataatac cctttggtaa 3421 tgtagcgctg tctccgtagt tttgcaaatc gcatttctcc agaagcattc tttgcatctt 3481 atacaaattg ggcatagcca caccaggctg ccaggcttga gaggacattt tgtaattaaa 3541 acttagatta gattgctatg ctttctttct aatgagcaag aagtaaaaaa agttgtaata 3601 gaacaagaaa aatgaaactg aaacttgaga aattgaagac cgtttattaa cttaaatatc 3661 aatgggaggt catcgaaaga gaaaaaaatc aaaaaaaaaa attttcaaga aaaagaaacg 3721 tgataaaaat ttttattgcc tttttcgacg aagaaaaaga aacgaggcgg tctctttttt 3781 cttttccaaa cctttagtac gggtaattaa cgacacccta gaggaagaaa gaggggaaat 3841 ttagtatgct gtgcttgggt gttttgaagt ggtacggcga tgcgcggagt ccgagaaaat 3901 ctggaagagt aaaaaaggag tagaaacatt ttgaagctat ggtgtgtggg gcggccgctc 3961 taactgacca tttaaatcat acctgacctc catagcagaa agtcaaaagc ctccgaccgg 4021 aggcttttga cttgatcggc acgtaagagg ttccaacttt caccataatg aaataagatc 4081 actaccgggc gtattttttg agttatcgag attttcagga gctaaggaag ctaaaatgag 4141 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt 4201 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt 4261 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagtttac gccccgaaga 4321 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat 4381 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga 4441 gtactcacca gtcacagaaa agcatctcac ggatggcatg acagtaagag aattatgcag 4501 tgctgccata accatgagtg ataacactgc ggccaactta cttctggcaa cgatcggagg 4561 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg 4621 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt 4681 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg 4741 gcaacaatta atagactgga tggaggcgga taaagttgca ggatcacttc tgcgctcggc 4801 cctcccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg 4861 tatcattgca gcactggggc cagatggtaa gccctcccgc atcgtagtta tctacacgac 4921 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact 4981 gattaagcat tggtaagtga ccaaacagga aaaaaccgcc cttaacatgg cccgctttat 5041 cagaagccag acattaacgc ttctggagaa actcaacgag ctggacgcgg atgaacaggc 5101 agacatctgt gaatcgcttc acgaccacgc tgatgagctt taccgcagct gcctcgcgcg 5161 tttcggtgat gacggtgaaa acctctgatg agggcccaaa tgtaatcacc tggctcacct 5221 tcgggtgggc ctttctgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca 5281 tcacaaaaat cgatgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca 5341 ggcgtttacc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg 5401 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag 5461 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt 5521 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca 5581 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg 5641 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt 5701 tggtatctgc gctctgctga agccagttac ctcggaaaaa gagttggtag ctcttgatcc 5761 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc 5821 agaaaaaaag gatctcaaga agatcctttg attttctacc gaagaaaggc ccacccgtga 5881 aggtgagcca gtgagttgat tgcagtccag ttacgctgga gtcaagcttg taacacgcgg 5941 ccgctcggga agttcctatt ctctagctag aaagtatagg aacttcagag cgcttttgaa 6001 aaccaaaagc gctctgatga cgcactttca aaaaaccaaa aacgcaccgg actgtaacga 6061 gctactaaaa tattgcgaat accgcttcca caaacattgc tcaaaagtat ctctttgcta 6121 tatatctctg tgctatatcc ctatataacc tacccatcca cctttcgctc cttgaacttg 6181 catctaaact cgacctctac attttttatg tttatctcta gtattactct ttagacaaaa 6241 aaattgtagt aagaactatt catagagtga atcgaaaaca atacgaaaat gtaaacattt 6301 cctatacgta gtatatagag acaaaataga agaaaccgtt cataattttc tgaccaatga 6361 agaatcatca acgctatcac tttctgttca caaagtatgc gcaatccaca tcggtataga 6421 atataatcgg ggatgccttt atcttgaaaa aatgcacccg cagcttcgct agtaatcagt 6481 aaacgcggga agtggagtca ggcttttttt atggaagaga aaatagacac caaagtagcc 6541 ttcttctaac cttaacggac ctacagtgca aaaagttatc aagagactgc attatagagc 6601 gcacaaagga gaaaaaaagt aatctaagat gctttgttag aaaaatagcg ctctcgggat 6661 gcatttttgt agaacaaaaa agaagtatag attctttgtt ggtaaaatag cgctctcgcg 6721 ttgcatttct gtt //