LOCUS Exported 7519 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Yeast expression plasmid for SARS-CoV-2 RNA-dependent RNA polymerase. ACCESSION . VERSION . KEYWORDS pGBW-m4046618 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7519) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 7519) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7519 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 10..417 /gene="S. cerevisiae TEF1" /label=TEF1 promoter /note="promoter for EF-1-alpha" primer_bind complement(3274..3292) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 3643..4446 /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 4529..5511 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 5589..5618 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 5640..6227 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 6239..6266 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(6290..6481) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(6482..7342) /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 7105..7124 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7343..7445) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 7446..7477 /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 atgtccgccg atgcacaatc tttcttaaat agagtctgcg gcgtgagtgc 481 tgctcgtcta accccttgtg gtacgggtac ttcaacagac gtagtttaca gggcgtttga 541 tatttataac gacaaggtcg ctggtttcgc taaatttttg aagaccaact gttgtagatt 601 ccaggagaag gatgaagacg acaatcttat cgattcgtac ttcgttgtta aaagacatac 661 tttttccaac tatcaacacg aagaaacaat atacaatttg ctcaaggact gtccagccgt 721 cgctaaacac gattttttca agttcagaat tgatggggac atggtgccac atatcagcag 781 acaacggttg accaagtaca ctatggctga tttagtttac gccttgcgac actttgacga 841 aggaaactgt gatactctga aggaaatttt ggttacctat aactgctgtg acgatgatta 901 cttcaacaaa aaggactggt acgacttcgt agagaatccc gacatcttga gggtttacgc 961 taacttaggt gaaagagtca gacaagcttt gttaaagaca gtccaatttt gcgatgctat 1021 gagaaatgcg ggtattgttg gtgttttgac tttggataat caagatttga acggtaactg 1081 gtatgacttc ggtgatttta tccagactac cccgggttct ggggtaccag ttgtcgattc 1141 ttactattct ctgttaatgc caatactcac cctaactaga gcattgactg ccgaatccca 1201 tgttgacact gatttaacta agccatacat taaatgggac cttctaaagt acgatttcac 1261 cgaagaacgt ttaaagttgt tcgacagata ttttaaatac tgggaccaaa cctaccaccc 1321 taattgtgtc aactgtttgg acgacagatg tattcttcac tgcgctaact ttaatgtgtt 1381 gttctcgact gtttttccac caacctcttt cggtccgctg gttcgaaaga tttttgttga 1441 cggtgttcca ttcgtggtca gtacgggata tcatttcaga gaattagggg tcgttcataa 1501 ccaagacgtg aacttgcact catcaagatt gtccttcaag gaattgttgg tctacgcagc 1561 tgatcctgca atgcacgccg cttctggaaa cttattgctg gataagagaa caacctgttt 1621 cagcgttgcc gctcttacta ataacgtcgc gttccaaact gtaaagccgg gtaactttaa 1681 taaagacttc tacgattttg ctgttagcaa gggttttttc aaagaaggtt ctagtgttga 1741 gttgaaacac tttttctttg ctcaagacgg caatgctgct atcagtgatt atgactatta 1801 ccgttacaac ttaccaacaa tgtgtgatat cagacagttg cttttcgtcg tcgaagttgt 1861 tgataaatat tttgactgtt atgatggtgg ttgcatcaat gcaaatcaag tcattgtcaa 1921 caacttagat aagtctgctg gcttcccatt caacaagtgg ggtaaagcca gattgtacta 1981 tgattccatg tcctacgagg accaagatgc tttgttcgcg tacaccaaga gaaacgttat 2041 tcctacaatt actcaaatga atctaaagta cgctatctct gcaaaaaatc gggccagaac 2101 cgttgctggt gtttctatct gttctacgat gactaaccgt caatttcatc agaagctatt 2161 gaaatccata gctgctacta gaggtgctac tgtagtcatc ggtacctcaa agttctatgg 2221 tggttggcac aatatgctca agaccgttta ctctgatgtc gaaaacccac atcttatggg 2281 ttgggactac cctaagtgtg atagagccat gccaaatatg ctaagaatta tggcatcctt 2341 ggtcttggct agaaagcata ccacatgctg ctcattgtcc caccgttttt acagattggc 2401 caacgaatgt gctcaagtcc tttctgaaat ggttatgtgt ggtggtagtc tatatgtaaa 2461 gccaggcgga acttcgtctg gagatgccac cactgcttac gctaactcgg tttttaacat 2521 ttgtcaagcc gttaccgcca acgtaaacgc tttgctgtct acggatggta acaagatcgc 2581 tgataagtat gtgcgcaact tgcaacatag actttatgaa tgtttgtata gaaatcgaga 2641 tgtcgatact gacttcgtta acgaatttta cgcgtactta agaaagcact tctccatgat 2701 gattttgtca gacgacgctg ttgtttgttt caattctact tacgctagcc aaggtttggt 2761 cgcttcaatc aaaaacttca agtctgtttt atactaccaa aacaatgttt ttatgtccga 2821 ggcaaaatgc tggacggaaa cagatttgac taagggtcca catgaatttt gtagtcaaca 2881 cacaatgtta gtgaaacaag gggacgacta cgtttaccta ccctacccag atccaagcag 2941 aattttgggt gccggttgtt tcgtggacga catagtcaag accgacggta ccttgatgat 3001 cgaacggttt gtctctctgg ccatagacgc ttacccattg actaagcacc ctaaccaaga 3061 atatgctgat gtttttcatt tatatcttca atacattaga aaattacatg acgaattgac 3121 cggtcacatg ttagatatgt actctgtgat gttgacgaac gataatacgt cacgttactg 3181 ggaacccgaa ttttacgaag cgatgtacac tccacacaca gtattgcagt aaacaggccc 3241 cttttccttt gtcgatatca tgtaattagt tatgtcacgc ttacattcac gccctcctcc 3301 cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta 3361 ttttttttaa tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt 3421 tttctgtaca aagcggccgc cgaaccacta gtgcactgca gtacacggtt tccttgaaat 3481 ttttttgatt cggtaatctc cgaacagaag gaagaacgaa ggaaggagca cagacttaga 3541 ttggtatata tacgcatatg tagtgttgaa gaaacatgaa attgcccagt attcttaacc 3601 caactgcaca gaacaaaaac gtgcaggaaa cgaagataaa tcatgtcgaa agctacatat 3661 aaggaacgtg ctgctactca tcctagtcct gttgctgcca agctatttaa tatcatgcac 3721 gaaaagcaaa caaacttgtg tgcttcattg gatgttcgta ccaccaagga attactggag 3781 ttagttgaag cattaggtcc caaaatttgt ttactaaaaa cacatgtgga tatcttgact 3841 gatttttcca tggagggcac agttaagccg ctaaaggcat tatccgccaa gtacaatttt 3901 ttactcttcg aggacagaaa atttgctgac attggtaata cagtcaaatt gcagtactct 3961 gcgggtgtat acagaatagc agaatgggca gacattacga atgcacacgg tgtggtgggc 4021 ccaggtattg ttagcggttt gaagcaggcg gcagaagaag taacaaagga acctagaggc 4081 cttttgatgt tagcagaatt gtcatgcaag ggctccctat ctactggaga atatactaag 4141 ggtactgttg acattgcgaa gagcgacaaa gattttgtta tcggctttat tgctcaaaga 4201 gacatgggtg gaagagatga aggttacgat tggttgatta tgacacccgg tgtgggttta 4261 gatgacaagg gagatgcatt gggtcaacag tatagaaccg tggatgatgt ggtttctaca 4321 ggatctgaca ttattattgt tggaagagga ctatttgcaa agggaaggga tgctaaggta 4381 gagggtgaac gttacagaaa agcaggctgg gaagcatatt tgagaagatg cggccagcaa 4441 aactaaaaaa ctgtattata agtaaatgca tgtatactaa actcacaaat tagagcttca 4501 atttaattat atcagttatt acccgagtaa cgaagcatct gtgcttcatt ttgtagaaca 4561 aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga 4621 acagaaatgc aacgcgaaag cgctatttta ccaacgaaga atctgtgctt catttttgta 4681 aaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt 4741 acagaacaga aatgcaacgc gagagcgcta ttttaccaac aaagaatcta tacttctttt 4801 ttgttctaca aaaatgcatc ccgagagcgc tatttttcta acaaagcatc ttagattact 4861 ttttttctcc tttgtgcgct ctataatgca gtctcttgat aactttttgc actgtaggtc 4921 cgttaaggtt agaagaaggc tactttggtg tctattttct cttccataaa aaaagcctga 4981 ctccacttcc cgcgtttact gattactagc gaagctgcgg gtgcattttt tcaagataaa 5041 ggcatccccg attatattct ataccgatgt ggattgcgca tactttgtga acagaaagtg 5101 atagcgttga tgattcttca ttggtcagaa aattatgaac ggtttcttct attttgtctc 5161 tatatactac gtataggaaa tgtttacatt ttcgtattgt tttcgattca ctctatgaat 5221 agttcttact acaatttttt tgtctaaaga gtaatactag agataaacat aaaaaatgta 5281 gaggtcgagt ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt tatataggga 5341 tatagcacag agatatatag caaagagata cttttgagca atgtttgtgg aagcggtatt 5401 cgcaatattt tagtagctcg ttacagtccg gtgcgttttt ggttttttga aagtgcgtca 5461 tcagagcgct tttggttttc aaaagcgctc tgaagttcct atactttcta gctagagaat 5521 aggaacttcc cgagcggccg cgtgttacaa gcttgactcc agcgtaactg gactgcaatc 5581 aactcactgg ctcaccttca cgggtgggcc tttcttcggt agaaaatcaa aggatcttct 5641 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca 5701 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaggta actggcttca 5761 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca 5821 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg 5881 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg 5941 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct 6001 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga 6061 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc 6121 ttccaggggt aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg 6181 agcatcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg 6241 cagaaaggcc cacccgaagg tgagccaggt gattacattt gggccctcat cagaggtttt 6301 caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa 6361 gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg 6421 ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca 6481 cttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca 6541 tagttgcctg actccccgtc gtgtagataa ctacgatgcg ggagggctta ccatctggcc 6601 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa 6661 accagccagc cgggagggcc gagcgcagaa gtgatcctgc aactttatcc gcctccatcc 6721 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca 6781 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat 6841 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag 6901 cggttagctc cttcggtcct ccgatcgttg ccagaagtaa gttggccgca gtgttatcac 6961 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgtg agatgctttt 7021 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt 7081 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc 7141 tcatcattgg aaaacgttct tcggggcgta aactctcaag gatcttaccg ctgttgagat 7201 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca 7261 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga 7321 cacggaaatg ttgaatactc attttagctt ccttagctcc tgaaaatctc gataactcaa 7381 aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc 7441 gatcaagtca aaagcctccg gtcggaggct tttgactttc tgctatggag gtcaggtatg 7501 atttaaatgg tcagttaga //