LOCUS Exported 6172 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Yeast Expression plasmid for SARS-CoV-2 nsp9. ACCESSION . VERSION . KEYWORDS pGBW-m4046558 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6172) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 6172) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6172 /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 767..784 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" primer_bind complement(1927..1945) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 2296..3099 /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 3182..4164 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 4242..4271 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 4293..4880 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 4892..4919 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(4943..5134) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(5135..5995) /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 5758..5777 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5996..6098) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 6099..6130 /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 atgaatgagc ttagtcctgt tgcactacga cagatgtctt gtgctgccgg 481 tactacacaa actgcttgca ctgatgacaa tgcgttagct tactacaaca caacaaaggg 541 aggtaggttt gtacttgcac tgttatccga tttacaggat ttgaaatggg ctagattccc 601 taagagtgat ggaactggta ctatctatac agaactggaa ccaccttgta ggtttgttac 661 agacacacct aaaggtccta aagtgaagta tttatacttt attaaaggat taaacaacct 721 aaatagaggt atggtacttg gtagtttagc tgccacagta cgtctactcg tgccgagagg 781 ttctaaaatc gaggaaggta agctagtcat ttggataaat ggagataagg gttacaacgg 841 tttggcagaa gtaggcaaga aatttgaaaa ggacacaggt atcaaggtga cggttgaaca 901 tccagacaag ttagaagaaa aattccctca agttgctgct actggtgatg gtcccgacat 961 tattttctgg gcccacgatc gttttggtgg atatgctcag tcaggtttgc tggctgagat 1021 caccccagac aaggccttcc aagataaatt gtacccattc acttgggatg ctgtcagata 1081 caacggtaag cttatagcat atccaattgc tgttgaagcc ttatcgttga tctacaataa 1141 ggacttgtta cctaacccac caaagacatg ggaagaaatt ccagctttgg ataaagagct 1201 aaaggccaag ggaaaaagcg ctttgatgtt taacttacaa gaaccttatt tcacctggcc 1261 attaatcgct gctgacggtg gttacgcatt taaatacgaa aacggtaagt atgacattaa 1321 ggatgtcggc gttgataatg ctggtgcaaa ggctggtttg accttcttgg ttgatttaat 1381 aaagaataaa cacatgaacg ctgacactga ctactctatc gccgaagccg ctttcaacaa 1441 gggtgaaact gctatgacca taaatggtcc atgggcctgg agtaacattg atacttctaa 1501 agtaaactat ggtgtcactg ttttgcctac attcaaaggc caaccatcca agccctttgt 1561 cggtgttcta tccgctggca tcaatgctgc ttctccaaac aaggagctag ctaaagaatt 1621 tttggaaaat tacttgttaa ctgacgaagg tttggaggcc gttaacaagg acaagccatt 1681 gggtgctgtc gctttgaaat cttacgaaga agaattggct aaggatccca gaattgccgc 1741 taccatggag aacgcccaaa agggtgaaat tatgccaaac atcccacaaa tgtcagcctt 1801 ctggtacgca gttaggactg ccgtgattaa tgctgcttcc ggtagacaaa ccgtcgacga 1861 agcattaaaa gatgctcaaa cgtaaacagg ccccttttcc tttgtcgata tcatgtaatt 1921 agttatgtca cgcttacatt cacgccctcc tcccacatcc gctctaaccg aaaaggaagg 1981 agttagacaa cctgaagtct aggtccctat ttattttttt taatagttat gttagtatta 2041 agaacgttat ttatatttca aatttttctt ttttttctgt acaaagcggc cgccgaacca 2101 ctagtgcact gcagtacacg gtttccttga aatttttttg attcggtaat ctccgaacag 2161 aaggaagaac gaaggaagga gcacagactt agattggtat atatacgcat atgtagtgtt 2221 gaagaaacat gaaattgccc agtattctta acccaactgc acagaacaaa aacgtgcagg 2281 aaacgaagat aaatcatgtc gaaagctaca tataaggaac gtgctgctac tcatcctagt 2341 cctgttgctg ccaagctatt taatatcatg cacgaaaagc aaacaaactt gtgtgcttca 2401 ttggatgttc gtaccaccaa ggaattactg gagttagttg aagcattagg tcccaaaatt 2461 tgtttactaa aaacacatgt ggatatcttg actgattttt ccatggaggg cacagttaag 2521 ccgctaaagg cattatccgc caagtacaat tttttactct tcgaggacag aaaatttgct 2581 gacattggta atacagtcaa attgcagtac tctgcgggtg tatacagaat agcagaatgg 2641 gcagacatta cgaatgcaca cggtgtggtg ggcccaggta ttgttagcgg tttgaagcag 2701 gcggcagaag aagtaacaaa ggaacctaga ggccttttga tgttagcaga attgtcatgc 2761 aagggctccc tatctactgg agaatatact aagggtactg ttgacattgc gaagagcgac 2821 aaagattttg ttatcggctt tattgctcaa agagacatgg gtggaagaga tgaaggttac 2881 gattggttga ttatgacacc cggtgtgggt ttagatgaca agggagatgc attgggtcaa 2941 cagtatagaa ccgtggatga tgtggtttct acaggatctg acattattat tgttggaaga 3001 ggactatttg caaagggaag ggatgctaag gtagagggtg aacgttacag aaaagcaggc 3061 tgggaagcat atttgagaag atgcggccag caaaactaaa aaactgtatt ataagtaaat 3121 gcatgtatac taaactcaca aattagagct tcaatttaat tatatcagtt attacccgag 3181 taacgaagca tctgtgcttc attttgtaga acaaaaatgc aacgcgagag cgctaatttt 3241 tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga aagcgctatt 3301 ttaccaacga agaatctgtg cttcattttt gtaaaacaaa aatgcaacgc gagagcgcta 3361 atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgagagcg 3421 ctattttacc aacaaagaat ctatacttct tttttgttct acaaaaatgc atcccgagag 3481 cgctattttt ctaacaaagc atcttagatt actttttttc tcctttgtgc gctctataat 3541 gcagtctctt gataactttt tgcactgtag gtccgttaag gttagaagaa ggctactttg 3601 gtgtctattt tctcttccat aaaaaaagcc tgactccact tcccgcgttt actgattact 3661 agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc ccgattatat tctataccga 3721 tgtggattgc gcatactttg tgaacagaaa gtgatagcgt tgatgattct tcattggtca 3781 gaaaattatg aacggtttct tctattttgt ctctatatac tacgtatagg aaatgtttac 3841 attttcgtat tgttttcgat tcactctatg aatagttctt actacaattt ttttgtctaa 3901 agagtaatac tagagataaa cataaaaaat gtagaggtcg agtttagatg caagttcaag 3961 gagcgaaagg tggatgggta ggttatatag ggatatagca cagagatata tagcaaagag 4021 atacttttga gcaatgtttg tggaagcggt attcgcaata ttttagtagc tcgttacagt 4081 ccggtgcgtt tttggttttt tgaaagtgcg tcatcagagc gcttttggtt ttcaaaagcg 4141 ctctgaagtt cctatacttt ctagctagag aataggaact tcccgagcgg ccgcgtgtta 4201 caagcttgac tccagcgtaa ctggactgca atcaactcac tggctcacct tcacgggtgg 4261 gcctttcttc ggtagaaaat caaaggatct tcttgagatc ctttttttct gcgcgtaatc 4321 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag 4381 ctaccaactc tttttccgag gtaactggct tcagcagagc gcagatacca aatactgttc 4441 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc 4501 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg 4561 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt 4621 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg 4681 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg 4741 gcagggtcgg aacaggagag cgcacgaggg agcttccagg ggtaaacgcc tggtatcttt 4801 atagtcctgt cgggtttcgc cacctctgac ttgagcatcg atttttgtga tgctcgtcag 4861 gggggcggag cctatggaaa aacgccagca acgcagaaag gcccacccga aggtgagcca 4921 ggtgattaca tttgggccct catcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc 4981 agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat 5041 ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg 5101 ccatgttaag ggcggttttt tcctgtttgg tcacttacca atgcttaatc agtgaggcac 5161 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga 5221 taactacgat gcgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc 5281 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccgggagg gccgagcgca 5341 gaagtgatcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta 5401 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg 5461 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc 5521 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg 5581 ttgccagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt 5641 ctcttactgt catgccatcc gtgagatgct tttctgtgac tggtgagtac tcaaccaagt 5701 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata 5761 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc 5821 gtaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac 5881 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa 5941 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcattttag 6001 cttccttagc tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt 6061 cattatggtg aaagttggaa cctcttacgt gccgatcaag tcaaaagcct ccggtcggag 6121 gcttttgact ttctgctatg gaggtcaggt atgatttaaa tggtcagtta ga //