LOCUS Exported 5092 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Yeast Expression plasmid for SARS-CoV-2 nsp9. ACCESSION . VERSION . KEYWORDS pGBW-m4046338 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5092) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 5092) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5092 /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" CDS 785..802 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(847..865) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 1216..2019 /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 2102..3084 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 3162..3191 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 3213..3800 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 3812..3839 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(3863..4054) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(4055..4915) /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 4678..4697 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4916..5018) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 5019..5050 /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 ttctcaccat catcaccatc attaaacagg ccccttttcc tttgtcgata tcatgtaatt 841 agttatgtca cgcttacatt cacgccctcc tcccacatcc gctctaaccg aaaaggaagg 901 agttagacaa cctgaagtct aggtccctat ttattttttt taatagttat gttagtatta 961 agaacgttat ttatatttca aatttttctt ttttttctgt acaaagcggc cgccgaacca 1021 ctagtgcact gcagtacacg gtttccttga aatttttttg attcggtaat ctccgaacag 1081 aaggaagaac gaaggaagga gcacagactt agattggtat atatacgcat atgtagtgtt 1141 gaagaaacat gaaattgccc agtattctta acccaactgc acagaacaaa aacgtgcagg 1201 aaacgaagat aaatcatgtc gaaagctaca tataaggaac gtgctgctac tcatcctagt 1261 cctgttgctg ccaagctatt taatatcatg cacgaaaagc aaacaaactt gtgtgcttca 1321 ttggatgttc gtaccaccaa ggaattactg gagttagttg aagcattagg tcccaaaatt 1381 tgtttactaa aaacacatgt ggatatcttg actgattttt ccatggaggg cacagttaag 1441 ccgctaaagg cattatccgc caagtacaat tttttactct tcgaggacag aaaatttgct 1501 gacattggta atacagtcaa attgcagtac tctgcgggtg tatacagaat agcagaatgg 1561 gcagacatta cgaatgcaca cggtgtggtg ggcccaggta ttgttagcgg tttgaagcag 1621 gcggcagaag aagtaacaaa ggaacctaga ggccttttga tgttagcaga attgtcatgc 1681 aagggctccc tatctactgg agaatatact aagggtactg ttgacattgc gaagagcgac 1741 aaagattttg ttatcggctt tattgctcaa agagacatgg gtggaagaga tgaaggttac 1801 gattggttga ttatgacacc cggtgtgggt ttagatgaca agggagatgc attgggtcaa 1861 cagtatagaa ccgtggatga tgtggtttct acaggatctg acattattat tgttggaaga 1921 ggactatttg caaagggaag ggatgctaag gtagagggtg aacgttacag aaaagcaggc 1981 tgggaagcat atttgagaag atgcggccag caaaactaaa aaactgtatt ataagtaaat 2041 gcatgtatac taaactcaca aattagagct tcaatttaat tatatcagtt attacccgag 2101 taacgaagca tctgtgcttc attttgtaga acaaaaatgc aacgcgagag cgctaatttt 2161 tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga aagcgctatt 2221 ttaccaacga agaatctgtg cttcattttt gtaaaacaaa aatgcaacgc gagagcgcta 2281 atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgagagcg 2341 ctattttacc aacaaagaat ctatacttct tttttgttct acaaaaatgc atcccgagag 2401 cgctattttt ctaacaaagc atcttagatt actttttttc tcctttgtgc gctctataat 2461 gcagtctctt gataactttt tgcactgtag gtccgttaag gttagaagaa ggctactttg 2521 gtgtctattt tctcttccat aaaaaaagcc tgactccact tcccgcgttt actgattact 2581 agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc ccgattatat tctataccga 2641 tgtggattgc gcatactttg tgaacagaaa gtgatagcgt tgatgattct tcattggtca 2701 gaaaattatg aacggtttct tctattttgt ctctatatac tacgtatagg aaatgtttac 2761 attttcgtat tgttttcgat tcactctatg aatagttctt actacaattt ttttgtctaa 2821 agagtaatac tagagataaa cataaaaaat gtagaggtcg agtttagatg caagttcaag 2881 gagcgaaagg tggatgggta ggttatatag ggatatagca cagagatata tagcaaagag 2941 atacttttga gcaatgtttg tggaagcggt attcgcaata ttttagtagc tcgttacagt 3001 ccggtgcgtt tttggttttt tgaaagtgcg tcatcagagc gcttttggtt ttcaaaagcg 3061 ctctgaagtt cctatacttt ctagctagag aataggaact tcccgagcgg ccgcgtgtta 3121 caagcttgac tccagcgtaa ctggactgca atcaactcac tggctcacct tcacgggtgg 3181 gcctttcttc ggtagaaaat caaaggatct tcttgagatc ctttttttct gcgcgtaatc 3241 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag 3301 ctaccaactc tttttccgag gtaactggct tcagcagagc gcagatacca aatactgttc 3361 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc 3421 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg 3481 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt 3541 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg 3601 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg 3661 gcagggtcgg aacaggagag cgcacgaggg agcttccagg ggtaaacgcc tggtatcttt 3721 atagtcctgt cgggtttcgc cacctctgac ttgagcatcg atttttgtga tgctcgtcag 3781 gggggcggag cctatggaaa aacgccagca acgcagaaag gcccacccga aggtgagcca 3841 ggtgattaca tttgggccct catcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc 3901 agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat 3961 ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg 4021 ccatgttaag ggcggttttt tcctgtttgg tcacttacca atgcttaatc agtgaggcac 4081 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga 4141 taactacgat gcgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc 4201 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccgggagg gccgagcgca 4261 gaagtgatcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta 4321 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg 4381 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc 4441 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg 4501 ttgccagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt 4561 ctcttactgt catgccatcc gtgagatgct tttctgtgac tggtgagtac tcaaccaagt 4621 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata 4681 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc 4741 gtaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac 4801 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa 4861 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcattttag 4921 cttccttagc tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt 4981 cattatggtg aaagttggaa cctcttacgt gccgatcaag tcaaaagcct ccggtcggag 5041 gcttttgact ttctgctatg gaggtcaggt atgatttaaa tggtcagtta ga //