LOCUS Exported 8635 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Yeast expression plasmid for SARS-CoV-2 RNA-dependent RNA polymerase. ACCESSION . VERSION . KEYWORDS pGBW-m4046219 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8635) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 8635) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8635 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(390..1193) /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 1544..1562 /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS complement(2705..2722) /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" promoter complement(5535..5942) /gene="S. cerevisiae TEF1" /label=TEF1 promoter /note="promoter for EF-1-alpha" terminator 5994..6025 /label=tonB terminator /note="bidirectional E. coli tonB-P14 transcription terminator" promoter 6026..6128 /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" CDS 6129..6989 /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(6347..6366) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" CDS 6990..7181 /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" terminator 7205..7232 /label=T7Te terminator /note="phage T7 early transcription terminator" rep_origin complement(7244..7831) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 7853..7882 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 7960..307 /label=2u ori /note="yeast 2u plasmid origin of replication" ORIGIN 1 aaatagcgct ctcgggatgc atttttgtag aacaaaaaag aagtatagat tctttgttgg 61 taaaatagcg ctctcgcgtt gcatttctgt tctgtaaaaa tgcagctcag attctttgtt 121 tgaaaaatta gcgctctcgc gttgcatttt tgttttacaa aaatgaagca cagattcttc 181 gttggtaaaa tagcgctttc gcgttgcatt tctgttctgt aaaaatgcag ctcagattct 241 ttgtttgaaa aattagcgct ctcgcgttgc atttttgttc tacaaaatga agcacagatg 301 cttcgttact cgggtaataa ctgatataat taaattgaag ctctaatttg tgagtttagt 361 atacatgcat ttacttataa tacagttttt tagttttgct ggccgcatct tctcaaatat 421 gcttcccagc ctgcttttct gtaacgttca ccctctacct tagcatccct tccctttgca 481 aatagtcctc ttccaacaat aataatgtca gatcctgtag aaaccacatc atccacggtt 541 ctatactgtt gacccaatgc atctcccttg tcatctaaac ccacaccggg tgtcataatc 601 aaccaatcgt aaccttcatc tcttccaccc atgtctcttt gagcaataaa gccgataaca 661 aaatctttgt cgctcttcgc aatgtcaaca gtacccttag tatattctcc agtagatagg 721 gagcccttgc atgacaattc tgctaacatc aaaaggcctc taggttcctt tgttacttct 781 tctgccgcct gcttcaaacc gctaacaata cctgggccca ccacaccgtg tgcattcgta 841 atgtctgccc attctgctat tctgtataca cccgcagagt actgcaattt gactgtatta 901 ccaatgtcag caaattttct gtcctcgaag agtaaaaaat tgtacttggc ggataatgcc 961 tttagcggct taactgtgcc ctccatggaa aaatcagtca agatatccac atgtgttttt 1021 agtaaacaaa ttttgggacc taatgcttca actaactcca gtaattcctt ggtggtacga 1081 acatccaatg aagcacacaa gtttgtttgc ttttcgtgca tgatattaaa tagcttggca 1141 gcaacaggac taggatgagt agcagcacgt tccttatatg tagctttcga catgatttat 1201 cttcgtttcc tgcacgtttt tgttctgtgc agttgggtta agaatactgg gcaatttcat 1261 gtttcttcaa cactacatat gcgtatatat accaatctaa gtctgtgctc cttccttcgt 1321 tcttccttct gttcggagat taccgaatca aaaaaatttc aaggaaaccg tgtactgcag 1381 tgcactagtg gttcggcggc cgctttgtac agaaaaaaaa gaaaaatttg aaatataaat 1441 aacgttctta atactaacat aactattaaa aaaaataaat agggacctag acttcaggtt 1501 gtctaactcc ttccttttcg gttagagcgg atgtgggagg agggcgtgaa tgtaagcgtg 1561 acataactaa ttacatgata tcgacaaagg aaaaggggcc tgtttacgtt tgagcatctt 1621 ttaatgcttc gtcgacggtt tgtctaccgg aagcagcatt aatcacggca gtcctaactg 1681 cgtaccagaa ggctgacatt tgtgggatgt ttggcataat ttcacccttt tgggcgttct 1741 ccatggtagc ggcaattctg ggatccttag ccaattcttc ttcgtaagat ttcaaagcga 1801 cagcacccaa tggcttgtcc ttgttaacgg cctccaaacc ttcgtcagtt aacaagtaat 1861 tttccaaaaa ttctttagct agctccttgt ttggagaagc agcattgatg ccagcggata 1921 gaacaccgac aaagggcttg gatggttggc ctttgaatgt aggcaaaaca gtgacaccat 1981 agtttacttt agaagtatca atgttactcc aggcccatgg accatttatg gtcatagcag 2041 tttcaccctt gttgaaagcg gcttcggcga tagagtagtc agtgtcagcg ttcatgtgtt 2101 tattctttat taaatcaacc aagaaggtca aaccagcctt tgcaccagca ttatcaacgc 2161 cgacatcctt aatgtcatac ttaccgtttt cgtatttaaa tgcgtaacca ccgtcagcag 2221 cgattaatgg ccaggtgaaa taaggttctt gtaagttaaa catcaaagcg ctttttccct 2281 tggcctttag ctctttatcc aaagctggaa tttcttccca tgtctttggt gggttaggta 2341 acaagtcctt attgtagatc aacgataagg cttcaacagc aattggatat gctataagct 2401 taccgttgta tctgacagca tcccaagtga atgggtacaa tttatcttgg aaggccttgt 2461 ctggggtgat ctcagccagc aaacctgact gagcatatcc accaaaacga tcgtgggccc 2521 agaaaataat gtcgggacca tcaccagtag cagcaacttg agggaatttt tcttctaact 2581 tgtctggatg ttcaaccgtc accttgatac ctgtgtcctt ttcaaatttc ttgcctactt 2641 ctgccaaacc gttgtaaccc ttatctccat ttatccaaat gactagctta ccttcctcga 2701 ttttagaacc tctcggcacg agctgcaata ctgtgtgtgg agtgtacatc gcttcgtaaa 2761 attcgggttc ccagtaacgt gacgtattat cgttcgtcaa catcacagag tacatatcta 2821 acatgtgacc ggtcaattcg tcatgtaatt ttctaatgta ttgaagatat aaatgaaaaa 2881 catcagcata ttcttggtta gggtgcttag tcaatgggta agcgtctatg gccagagaga 2941 caaaccgttc gatcatcaag gtaccgtcgg tcttgactat gtcgtccacg aaacaaccgg 3001 cacccaaaat tctgcttgga tctgggtagg gtaggtaaac gtagtcgtcc ccttgtttca 3061 ctaacattgt gtgttgacta caaaattcat gtggaccctt agtcaaatct gtttccgtcc 3121 agcattttgc ctcggacata aaaacattgt tttggtagta taaaacagac ttgaagtttt 3181 tgattgaagc gaccaaacct tggctagcgt aagtagaatt gaaacaaaca acagcgtcgt 3241 ctgacaaaat catcatggag aagtgctttc ttaagtacgc gtaaaattcg ttaacgaagt 3301 cagtatcgac atctcgattt ctatacaaac attcataaag tctatgttgc aagttgcgca 3361 catacttatc agcgatcttg ttaccatccg tagacagcaa agcgtttacg ttggcggtaa 3421 cggcttgaca aatgttaaaa accgagttag cgtaagcagt ggtggcatct ccagacgaag 3481 ttccgcctgg ctttacatat agactaccac cacacataac catttcagaa aggacttgag 3541 cacattcgtt ggccaatctg taaaaacggt gggacaatga gcagcatgtg gtatgctttc 3601 tagccaagac caaggatgcc ataattctta gcatatttgg catggctcta tcacacttag 3661 ggtagtccca acccataaga tgtgggtttt cgacatcaga gtaaacggtc ttgagcatat 3721 tgtgccaacc accatagaac tttgaggtac cgatgactac agtagcacct ctagtagcag 3781 ctatggattt caatagcttc tgatgaaatt gacggttagt catcgtagaa cagatagaaa 3841 caccagcaac ggttctggcc cgattttttg cagagatagc gtactttaga ttcatttgag 3901 taattgtagg aataacgttt ctcttggtgt acgcgaacaa agcatcttgg tcctcgtagg 3961 acatggaatc atagtacaat ctggctttac cccacttgtt gaatgggaag ccagcagact 4021 tatctaagtt gttgacaatg acttgatttg cattgatgca accaccatca taacagtcaa 4081 aatatttatc aacaacttcg acgacgaaaa gcaactgtct gatatcacac attgttggta 4141 agttgtaacg gtaatagtca taatcactga tagcagcatt gccgtcttga gcaaagaaaa 4201 agtgtttcaa ctcaacacta gaaccttctt tgaaaaaacc cttgctaaca gcaaaatcgt 4261 agaagtcttt attaaagtta cccggcttta cagtttggaa cgcgacgtta ttagtaagag 4321 cggcaacgct gaaacaggtt gttctcttat ccagcaataa gtttccagaa gcggcgtgca 4381 ttgcaggatc agctgcgtag accaacaatt ccttgaagga caatcttgat gagtgcaagt 4441 tcacgtcttg gttatgaacg acccctaatt ctctgaaatg atatcccgta ctgaccacga 4501 atggaacacc gtcaacaaaa atctttcgaa ccagcggacc gaaagaggtt ggtggaaaaa 4561 cagtcgagaa caacacatta aagttagcgc agtgaagaat acatctgtcg tccaaacagt 4621 tgacacaatt agggtggtag gtttggtccc agtatttaaa atatctgtcg aacaacttta 4681 aacgttcttc ggtgaaatcg tactttagaa ggtcccattt aatgtatggc ttagttaaat 4741 cagtgtcaac atgggattcg gcagtcaatg ctctagttag ggtgagtatt ggcattaaca 4801 gagaatagta agaatcgaca actggtaccc cagaacccgg ggtagtctgg ataaaatcac 4861 cgaagtcata ccagttaccg ttcaaatctt gattatccaa agtcaaaaca ccaacaatac 4921 ccgcatttct catagcatcg caaaattgga ctgtctttaa caaagcttgt ctgactcttt 4981 cacctaagtt agcgtaaacc ctcaagatgt cgggattctc tacgaagtcg taccagtcct 5041 ttttgttgaa gtaatcatcg tcacagcagt tataggtaac caaaatttcc ttcagagtat 5101 cacagtttcc ttcgtcaaag tgtcgcaagg cgtaaactaa atcagccata gtgtacttgg 5161 tcaaccgttg tctgctgata tgtggcacca tgtccccatc aattctgaac ttgaaaaaat 5221 cgtgtttagc gacggctgga cagtccttga gcaaattgta tattgtttct tcgtgttgat 5281 agttggaaaa agtatgtctt ttaacaacga agtacgaatc gataagattg tcgtcttcat 5341 ccttctcctg gaatctacaa cagttggtct tcaaaaattt agcgaaacca gcgaccttgt 5401 cgttataaat atcaaacgcc ctgtaaacta cgtctgttga agtacccgta ccacaagggg 5461 ttagacgagc agcactcacg ccgcagactc tatttaagaa agattgtgca tcggcggaca 5521 ttttgtaatt aaaacttaga ttagattgct atgctttctt tctaatgagc aagaagtaaa 5581 aaaagttgta atagaacaag aaaaatgaaa ctgaaacttg agaaattgaa gaccgtttat 5641 taacttaaat atcaatggga ggtcatcgaa agagaaaaaa atcaaaaaaa aaaattttca 5701 agaaaaagaa acgtgataaa aatttttatt gcctttttcg acgaagaaaa agaaacgagg 5761 cggtctcttt tttcttttcc aaacctttag tacgggtaat taacgacacc ctagaggaag 5821 aaagagggga aatttagtat gctgtgcttg ggtgttttga agtggtacgg cgatgcgcgg 5881 agtccgagaa aatctggaag agtaaaaaag gagtagaaac attttgaagc tatggtgtgt 5941 ggggcggccg ctctaactga ccatttaaat catacctgac ctccatagca gaaagtcaaa 6001 agcctccgac cggaggcttt tgacttgatc ggcacgtaag aggttccaac tttcaccata 6061 atgaaataag atcactaccg ggcgtatttt ttgagttatc gagattttca ggagctaagg 6121 aagctaaaat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt 6181 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt 6241 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt 6301 tacgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg 6361 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga 6421 atgacttggt tgagtactca ccagtcacag aaaagcatct cacggatggc atgacagtaa 6481 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctgg 6541 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa 6601 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca 6661 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta 6721 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggatcac 6781 ttctgcgctc ggccctcccg gctggctggt ttattgctga taaatctgga gccggtgagc 6841 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgcatcgtag 6901 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga 6961 taggtgcctc actgattaag cattggtaag tgaccaaaca ggaaaaaacc gcccttaaca 7021 tggcccgctt tatcagaagc cagacattaa cgcttctgga gaaactcaac gagctggacg 7081 cggatgaaca ggcagacatc tgtgaatcgc ttcacgacca cgctgatgag ctttaccgca 7141 gctgcctcgc gcgtttcggt gatgacggtg aaaacctctg atgagggccc aaatgtaatc 7201 acctggctca ccttcgggtg ggcctttctg cgttgctggc gtttttccat aggctccgcc 7261 cccctgacga gcatcacaaa aatcgatgct caagtcagag gtggcgaaac ccgacaggac 7321 tataaagata ccaggcgttt acccctggaa gctccctcgt gcgctctcct gttccgaccc 7381 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata 7441 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc 7501 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca 7561 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag 7621 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta 7681 gaagaacagt atttggtatc tgcgctctgc tgaagccagt tacctcggaa aaagagttgg 7741 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 7801 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgattttct accgaagaaa 7861 ggcccacccg tgaaggtgag ccagtgagtt gattgcagtc cagttacgct ggagtcaagc 7921 ttgtaacacg cggccgctcg ggaagttcct attctctagc tagaaagtat aggaacttca 7981 gagcgctttt gaaaaccaaa agcgctctga tgacgcactt tcaaaaaacc aaaaacgcac 8041 cggactgtaa cgagctacta aaatattgcg aataccgctt ccacaaacat tgctcaaaag 8101 tatctctttg ctatatatct ctgtgctata tccctatata acctacccat ccacctttcg 8161 ctccttgaac ttgcatctaa actcgacctc tacatttttt atgtttatct ctagtattac 8221 tctttagaca aaaaaattgt agtaagaact attcatagag tgaatcgaaa acaatacgaa 8281 aatgtaaaca tttcctatac gtagtatata gagacaaaat agaagaaacc gttcataatt 8341 ttctgaccaa tgaagaatca tcaacgctat cactttctgt tcacaaagta tgcgcaatcc 8401 acatcggtat agaatataat cggggatgcc tttatcttga aaaaatgcac ccgcagcttc 8461 gctagtaatc agtaaacgcg ggaagtggag tcaggctttt tttatggaag agaaaataga 8521 caccaaagta gccttcttct aaccttaacg gacctacagt gcaaaaagtt atcaagagac 8581 tgcattatag agcgcacaaa ggagaaaaaa agtaatctaa gatgctttgt tagaa //