LOCUS Exported 6774 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Yeast Expression plasmid for SARS-CoV-2 2'-O-ribose methyltransferase. ACCESSION . VERSION . KEYWORDS pGBW-m4046487 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6774) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 6774) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6774 /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 1369..1386 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" primer_bind complement(2529..2547) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 2898..3701 /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 3784..4766 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 4844..4873 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 4895..5482 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 5494..5521 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(5545..5736) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(5737..6597) /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 6360..6379 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6598..6700) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 6701..6732 /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 agtcaagcgt ggcaaccggg tgttgctatg cctaatcttt acaaaatgca aagaatgcta 541 ttagaaaagt gtgaccttca aaattatggt gatagtgcaa cattacctaa aggcataatg 601 atgaatgtcg caaaatatac tcaactgtgt caatatttaa acacattaac attagctgta 661 ccctataata tgagagttat acattttggt gctggttctg ataaaggagt tgcaccaggt 721 acagctgttt taagacagtg gttgcctacg ggtacgctgc ttgtcgattc agatcttaat 781 gactttgtct ctgatgcaga ttcaactttg attggtgatt gtgcaactgt acatacagct 841 aataaatggg atctcattat tagtgatatg tacgacccta agactaaaaa tgttacaaaa 901 gaaaatgact ctaaagaggg ttttttcact tacatttgtg ggtttataca acaaaagcta 961 gctcttggag gttccgtggc tataaagata acagaacatt cttggaatgc tgatctttat 1021 aagctcatgg gacacttcgc atggtggaca gcctttgtta ctaatgtgaa tgcgtcatca 1081 tctgaagcat ttttaattgg atgtaattat cttggcaaac cacgcgaaca aatagatggt 1141 tatgtcatgc atgcaaatta catattttgg aggaatacaa atccaattca gttgtcttcc 1201 tattctttat ttgacatgag taaatttccc cttaaattaa ggggtactgc tgttatgtct 1261 ttaaaagaag gtcaaatcaa tgatatgatt ttatctcttc ttagtaaagg tagacttata 1321 attagagaaa acaacagagt tgttatttct agtgatgttc ttgttaacct cgtgccgaga 1381 ggttctaaaa tcgaggaagg taagctagtc atttggataa atggagataa gggttacaac 1441 ggtttggcag aagtaggcaa gaaatttgaa aaggacacag gtatcaaggt gacggttgaa 1501 catccagaca agttagaaga aaaattccct caagttgctg ctactggtga tggtcccgac 1561 attattttct gggcccacga tcgttttggt ggatatgctc agtcaggttt gctggctgag 1621 atcaccccag acaaggcctt ccaagataaa ttgtacccat tcacttggga tgctgtcaga 1681 tacaacggta agcttatagc atatccaatt gctgttgaag ccttatcgtt gatctacaat 1741 aaggacttgt tacctaaccc accaaagaca tgggaagaaa ttccagcttt ggataaagag 1801 ctaaaggcca agggaaaaag cgctttgatg tttaacttac aagaacctta tttcacctgg 1861 ccattaatcg ctgctgacgg tggttacgca tttaaatacg aaaacggtaa gtatgacatt 1921 aaggatgtcg gcgttgataa tgctggtgca aaggctggtt tgaccttctt ggttgattta 1981 ataaagaata aacacatgaa cgctgacact gactactcta tcgccgaagc cgctttcaac 2041 aagggtgaaa ctgctatgac cataaatggt ccatgggcct ggagtaacat tgatacttct 2101 aaagtaaact atggtgtcac tgttttgcct acattcaaag gccaaccatc caagcccttt 2161 gtcggtgttc tatccgctgg catcaatgct gcttctccaa acaaggagct agctaaagaa 2221 tttttggaaa attacttgtt aactgacgaa ggtttggagg ccgttaacaa ggacaagcca 2281 ttgggtgctg tcgctttgaa atcttacgaa gaagaattgg ctaaggatcc cagaattgcc 2341 gctaccatgg agaacgccca aaagggtgaa attatgccaa acatcccaca aatgtcagcc 2401 ttctggtacg cagttaggac tgccgtgatt aatgctgctt ccggtagaca aaccgtcgac 2461 gaagcattaa aagatgctca aacgtaaaca ggcccctttt cctttgtcga tatcatgtaa 2521 ttagttatgt cacgcttaca ttcacgccct cctcccacat ccgctctaac cgaaaaggaa 2581 ggagttagac aacctgaagt ctaggtccct atttattttt tttaatagtt atgttagtat 2641 taagaacgtt atttatattt caaatttttc ttttttttct gtacaaagcg gccgccgaac 2701 cactagtgca ctgcagtaca cggtttcctt gaaatttttt tgattcggta atctccgaac 2761 agaaggaaga acgaaggaag gagcacagac ttagattggt atatatacgc atatgtagtg 2821 ttgaagaaac atgaaattgc ccagtattct taacccaact gcacagaaca aaaacgtgca 2881 ggaaacgaag ataaatcatg tcgaaagcta catataagga acgtgctgct actcatccta 2941 gtcctgttgc tgccaagcta tttaatatca tgcacgaaaa gcaaacaaac ttgtgtgctt 3001 cattggatgt tcgtaccacc aaggaattac tggagttagt tgaagcatta ggtcccaaaa 3061 tttgtttact aaaaacacat gtggatatct tgactgattt ttccatggag ggcacagtta 3121 agccgctaaa ggcattatcc gccaagtaca attttttact cttcgaggac agaaaatttg 3181 ctgacattgg taatacagtc aaattgcagt actctgcggg tgtatacaga atagcagaat 3241 gggcagacat tacgaatgca cacggtgtgg tgggcccagg tattgttagc ggtttgaagc 3301 aggcggcaga agaagtaaca aaggaaccta gaggcctttt gatgttagca gaattgtcat 3361 gcaagggctc cctatctact ggagaatata ctaagggtac tgttgacatt gcgaagagcg 3421 acaaagattt tgttatcggc tttattgctc aaagagacat gggtggaaga gatgaaggtt 3481 acgattggtt gattatgaca cccggtgtgg gtttagatga caagggagat gcattgggtc 3541 aacagtatag aaccgtggat gatgtggttt ctacaggatc tgacattatt attgttggaa 3601 gaggactatt tgcaaaggga agggatgcta aggtagaggg tgaacgttac agaaaagcag 3661 gctgggaagc atatttgaga agatgcggcc agcaaaacta aaaaactgta ttataagtaa 3721 atgcatgtat actaaactca caaattagag cttcaattta attatatcag ttattacccg 3781 agtaacgaag catctgtgct tcattttgta gaacaaaaat gcaacgcgag agcgctaatt 3841 tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc gaaagcgcta 3901 ttttaccaac gaagaatctg tgcttcattt ttgtaaaaca aaaatgcaac gcgagagcgc 3961 taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgagag 4021 cgctatttta ccaacaaaga atctatactt cttttttgtt ctacaaaaat gcatcccgag 4081 agcgctattt ttctaacaaa gcatcttaga ttactttttt tctcctttgt gcgctctata 4141 atgcagtctc ttgataactt tttgcactgt aggtccgtta aggttagaag aaggctactt 4201 tggtgtctat tttctcttcc ataaaaaaag cctgactcca cttcccgcgt ttactgatta 4261 ctagcgaagc tgcgggtgca ttttttcaag ataaaggcat ccccgattat attctatacc 4321 gatgtggatt gcgcatactt tgtgaacaga aagtgatagc gttgatgatt cttcattggt 4381 cagaaaatta tgaacggttt cttctatttt gtctctatat actacgtata ggaaatgttt 4441 acattttcgt attgttttcg attcactcta tgaatagttc ttactacaat ttttttgtct 4501 aaagagtaat actagagata aacataaaaa atgtagaggt cgagtttaga tgcaagttca 4561 aggagcgaaa ggtggatggg taggttatat agggatatag cacagagata tatagcaaag 4621 agatactttt gagcaatgtt tgtggaagcg gtattcgcaa tattttagta gctcgttaca 4681 gtccggtgcg tttttggttt tttgaaagtg cgtcatcaga gcgcttttgg ttttcaaaag 4741 cgctctgaag ttcctatact ttctagctag agaataggaa cttcccgagc ggccgcgtgt 4801 tacaagcttg actccagcgt aactggactg caatcaactc actggctcac cttcacgggt 4861 gggcctttct tcggtagaaa atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa 4921 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag 4981 agctaccaac tctttttccg aggtaactgg cttcagcaga gcgcagatac caaatactgt 5041 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata 5101 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac 5161 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg 5221 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg 5281 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag 5341 cggcagggtc ggaacaggag agcgcacgag ggagcttcca ggggtaaacg cctggtatct 5401 ttatagtcct gtcgggtttc gccacctctg acttgagcat cgatttttgt gatgctcgtc 5461 aggggggcgg agcctatgga aaaacgccag caacgcagaa aggcccaccc gaaggtgagc 5521 caggtgatta catttgggcc ctcatcagag gttttcaccg tcatcaccga aacgcgcgag 5581 gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc 5641 atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg 5701 ggccatgtta agggcggttt tttcctgttt ggtcacttac caatgcttaa tcagtgaggc 5761 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta 5821 gataactacg atgcgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga 5881 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggga gggccgagcg 5941 cagaagtgat cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc 6001 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat 6061 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag 6121 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat 6181 cgttgccaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa 6241 ttctcttact gtcatgccat ccgtgagatg cttttctgtg actggtgagt actcaaccaa 6301 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga 6361 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg 6421 gcgtaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc 6481 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg 6541 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatttt 6601 agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat 6661 ttcattatgg tgaaagttgg aacctcttac gtgccgatca agtcaaaagc ctccggtcgg 6721 aggcttttga ctttctgcta tggaggtcag gtatgattta aatggtcagt taga //