LOCUS Exported 7689 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Yeast Expression plasmid for SARS-CoV-2 helicase. ACCESSION . VERSION . KEYWORDS pGBW-m4046552 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7689) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 7689) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7689 /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 2284..2301 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" primer_bind complement(3444..3462) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 3813..4616 /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 4699..5681 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 5759..5788 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 5810..6397 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 6409..6436 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(6460..6651) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(6652..7512) /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 7275..7294 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7513..7615) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 7616..7647 /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 gcagtcggcg cttgtgtatt atgcaactcc cagacgtcgt tgagatgtgg tgcttgtatt 541 cgtagacctt tcctttgctg taaatgttgc tatgaccacg ttatctcaac ttctcataag 601 ctagttctga gcgtcaatcc atacgtgtgt aacgcccccg ggtgtgatgt tacagacgtt 661 acccaattgt acttgggagg tatgtcttac tattgtaagt cccacaagcc acctatatct 721 tttccattat gcgctaacgg tcaagtcttc ggtttgtaca aaaatacctg tgttggtagt 781 gataatgtta ccgactttaa cgccattgct acttgtgatt ggactaacgc gggtgattat 841 atcttggcta acacttgtac cgagaggctt aagttgttcg ccgctgaaac cttaaaggct 901 acggaagaaa cattcaaatt gagttacggt atcgctactg tcagagaagt tctatctgat 961 agagaattgc atctctcctg ggaggtgggt aagccaagac caccgttgaa ccggaattac 1021 gtctttactg gatacagggt tactaagaac tctaaggttc aaattgggga atataccttt 1081 gaaaagggcg actacggtga cgccgtagtc tatagaggta ccaccactta caaattgaac 1141 gtcggtgact atttcgtttt aaccagccac acagtgatgc cactgtcagc gccaacatta 1201 gttccgcaag agcattacgt tagaattact ggtttgtatc caactctaaa tatctctgat 1261 gaattttcct ccaatgtggc taactaccaa aaagtaggta tgcaaaagta ctctaccttg 1321 caaggtccac ctggtactgg gaagtcgcac ttcgctatag gtttggcact gtactaccca 1381 tctgctagaa ttgtttacac ggcatgttct catgctgctg tagacgcctt gtgtgaaaag 1441 gccttgaaat atttacccat tgataaatgt tcaagaatta tcccagcacg agctcgtgtt 1501 gaatgtttcg acaagtttaa ggtcaacagc acactagaac aatacgtctt ctgcactgtt 1561 aatgctttgc cagaaacgac tgctgatatc gtcgtctttg atgaaatctc gatggccact 1621 aactatgact tatccgtagt taatgctaga cttcgtgcaa agcactacgt ctatattggt 1681 gatccagctc agctgcctgc accaagaact ttgttaacca agggtacctt ggaaccagaa 1741 tacttcaact ctgtgtgtcg cctgatgaag acaatcggcc ctgatatgtt cctaggtaca 1801 tgtagacgtt gtccagctga aattgtggac acggtcagtg ccttggttta cgataacaaa 1861 ttaaaggcgc ataaggacaa aagcgctcaa tgtttcaaga tgttttacaa gggtgtcatt 1921 actcacgatg ttagttcggc catcaataga ccccaaatag gtgttgttag agaatttttg 1981 acaaggaacc cagcatggag aaaggctgtt ttcatatccc catacaattc tcaaaacgcg 2041 gtagcctcaa aaatcttggg tcttcccact cagaccgttg attcttccca aggaagtgaa 2101 tatgactacg tgattttcac tcaaaccacc gagactgccc actcttgcaa cgtcaatcgt 2161 ttcaacgttg ctattacgag agccaaggtt ggcatcttgt gtatcatgtc tgatagagac 2221 ttgtacgata aattacagtt tacatcctta gaaataccaa gacgaaacgt cgctactttg 2281 caactcgtgc cgagaggttc taaaatcgag gaaggtaagc tagtcatttg gataaatgga 2341 gataagggtt acaacggttt ggcagaagta ggcaagaaat ttgaaaagga cacaggtatc 2401 aaggtgacgg ttgaacatcc agacaagtta gaagaaaaat tccctcaagt tgctgctact 2461 ggtgatggtc ccgacattat tttctgggcc cacgatcgtt ttggtggata tgctcagtca 2521 ggtttgctgg ctgagatcac cccagacaag gccttccaag ataaattgta cccattcact 2581 tgggatgctg tcagatacaa cggtaagctt atagcatatc caattgctgt tgaagcctta 2641 tcgttgatct acaataagga cttgttacct aacccaccaa agacatggga agaaattcca 2701 gctttggata aagagctaaa ggccaaggga aaaagcgctt tgatgtttaa cttacaagaa 2761 ccttatttca cctggccatt aatcgctgct gacggtggtt acgcatttaa atacgaaaac 2821 ggtaagtatg acattaagga tgtcggcgtt gataatgctg gtgcaaaggc tggtttgacc 2881 ttcttggttg atttaataaa gaataaacac atgaacgctg acactgacta ctctatcgcc 2941 gaagccgctt tcaacaaggg tgaaactgct atgaccataa atggtccatg ggcctggagt 3001 aacattgata cttctaaagt aaactatggt gtcactgttt tgcctacatt caaaggccaa 3061 ccatccaagc cctttgtcgg tgttctatcc gctggcatca atgctgcttc tccaaacaag 3121 gagctagcta aagaattttt ggaaaattac ttgttaactg acgaaggttt ggaggccgtt 3181 aacaaggaca agccattggg tgctgtcgct ttgaaatctt acgaagaaga attggctaag 3241 gatcccagaa ttgccgctac catggagaac gcccaaaagg gtgaaattat gccaaacatc 3301 ccacaaatgt cagccttctg gtacgcagtt aggactgccg tgattaatgc tgcttccggt 3361 agacaaaccg tcgacgaagc attaaaagat gctcaaacgt aaacaggccc cttttccttt 3421 gtcgatatca tgtaattagt tatgtcacgc ttacattcac gccctcctcc cacatccgct 3481 ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta ttttttttaa 3541 tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt tttctgtaca 3601 aagcggccgc cgaaccacta gtgcactgca gtacacggtt tccttgaaat ttttttgatt 3661 cggtaatctc cgaacagaag gaagaacgaa ggaaggagca cagacttaga ttggtatata 3721 tacgcatatg tagtgttgaa gaaacatgaa attgcccagt attcttaacc caactgcaca 3781 gaacaaaaac gtgcaggaaa cgaagataaa tcatgtcgaa agctacatat aaggaacgtg 3841 ctgctactca tcctagtcct gttgctgcca agctatttaa tatcatgcac gaaaagcaaa 3901 caaacttgtg tgcttcattg gatgttcgta ccaccaagga attactggag ttagttgaag 3961 cattaggtcc caaaatttgt ttactaaaaa cacatgtgga tatcttgact gatttttcca 4021 tggagggcac agttaagccg ctaaaggcat tatccgccaa gtacaatttt ttactcttcg 4081 aggacagaaa atttgctgac attggtaata cagtcaaatt gcagtactct gcgggtgtat 4141 acagaatagc agaatgggca gacattacga atgcacacgg tgtggtgggc ccaggtattg 4201 ttagcggttt gaagcaggcg gcagaagaag taacaaagga acctagaggc cttttgatgt 4261 tagcagaatt gtcatgcaag ggctccctat ctactggaga atatactaag ggtactgttg 4321 acattgcgaa gagcgacaaa gattttgtta tcggctttat tgctcaaaga gacatgggtg 4381 gaagagatga aggttacgat tggttgatta tgacacccgg tgtgggttta gatgacaagg 4441 gagatgcatt gggtcaacag tatagaaccg tggatgatgt ggtttctaca ggatctgaca 4501 ttattattgt tggaagagga ctatttgcaa agggaaggga tgctaaggta gagggtgaac 4561 gttacagaaa agcaggctgg gaagcatatt tgagaagatg cggccagcaa aactaaaaaa 4621 ctgtattata agtaaatgca tgtatactaa actcacaaat tagagcttca atttaattat 4681 atcagttatt acccgagtaa cgaagcatct gtgcttcatt ttgtagaaca aaaatgcaac 4741 gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc 4801 aacgcgaaag cgctatttta ccaacgaaga atctgtgctt catttttgta aaacaaaaat 4861 gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga 4921 aatgcaacgc gagagcgcta ttttaccaac aaagaatcta tacttctttt ttgttctaca 4981 aaaatgcatc ccgagagcgc tatttttcta acaaagcatc ttagattact ttttttctcc 5041 tttgtgcgct ctataatgca gtctcttgat aactttttgc actgtaggtc cgttaaggtt 5101 agaagaaggc tactttggtg tctattttct cttccataaa aaaagcctga ctccacttcc 5161 cgcgtttact gattactagc gaagctgcgg gtgcattttt tcaagataaa ggcatccccg 5221 attatattct ataccgatgt ggattgcgca tactttgtga acagaaagtg atagcgttga 5281 tgattcttca ttggtcagaa aattatgaac ggtttcttct attttgtctc tatatactac 5341 gtataggaaa tgtttacatt ttcgtattgt tttcgattca ctctatgaat agttcttact 5401 acaatttttt tgtctaaaga gtaatactag agataaacat aaaaaatgta gaggtcgagt 5461 ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt tatataggga tatagcacag 5521 agatatatag caaagagata cttttgagca atgtttgtgg aagcggtatt cgcaatattt 5581 tagtagctcg ttacagtccg gtgcgttttt ggttttttga aagtgcgtca tcagagcgct 5641 tttggttttc aaaagcgctc tgaagttcct atactttcta gctagagaat aggaacttcc 5701 cgagcggccg cgtgttacaa gcttgactcc agcgtaactg gactgcaatc aactcactgg 5761 ctcaccttca cgggtgggcc tttcttcggt agaaaatcaa aggatcttct tgagatcctt 5821 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt 5881 gtttgccgga tcaagagcta ccaactcttt ttccgaggta actggcttca gcagagcgca 5941 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt 6001 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga 6061 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc 6121 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact 6181 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga 6241 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggt 6301 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcatcgatt 6361 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cagaaaggcc 6421 cacccgaagg tgagccaggt gattacattt gggccctcat cagaggtttt caccgtcatc 6481 accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca 6541 gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg 6601 gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca cttaccaatg 6661 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg 6721 actccccgtc gtgtagataa ctacgatgcg ggagggctta ccatctggcc ccagtgctgc 6781 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc 6841 cgggagggcc gagcgcagaa gtgatcctgc aactttatcc gcctccatcc agtctattaa 6901 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc 6961 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg 7021 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc 7081 cttcggtcct ccgatcgttg ccagaagtaa gttggccgca gtgttatcac tcatggttat 7141 ggcagcactg cataattctc ttactgtcat gccatccgtg agatgctttt ctgtgactgg 7201 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc 7261 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg 7321 aaaacgttct tcggggcgta aactctcaag gatcttaccg ctgttgagat ccagttcgat 7381 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg 7441 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg 7501 ttgaatactc attttagctt ccttagctcc tgaaaatctc gataactcaa aaaatacgcc 7561 cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc gatcaagtca 7621 aaagcctccg gtcggaggct tttgactttc tgctatggag gtcaggtatg atttaaatgg 7681 tcagttaga //