LOCUS Exported 6609 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Yeast Expression plasmid for SARS-CoV-2 helicase. ACCESSION . VERSION . KEYWORDS pGBW-m4046305 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6609) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 6609) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6609 /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 2278..2295 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" CDS 2296..2319 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" primer_bind complement(2364..2382) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 2733..3536 /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 3619..4601 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 4679..4708 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 4730..5317 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 5329..5356 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(5380..5571) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(5572..6432) /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 6195..6214 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6433..6535) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 6536..6567 /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 gttggggctt gtgttctttg caattcacag acttcattaa gatgtggtgc ttgcatacgt 541 agaccattct tatgttgtaa atgctgttac gaccatgtca tatcaacatc acataaatta 601 gtcttgtctg ttaatccgta tgtttgcaat gctccaggtt gtgatgtcac agatgtgact 661 caactttact taggaggtat gagctattat tgtaaatcac ataaaccacc cattagtttt 721 ccattgtgtg ctaatggaca agtttttggt ttatataaaa atacatgtgt tggtagcgat 781 aatgttactg actttaatgc aattgcaaca tgtgactgga caaatgctgg tgattacatt 841 ttagctaaca cctgtactga aagactcaag ctttttgcag cagaaacgct caaagctact 901 gaggagacat ttaaactgtc ttatggtatt gctactgtac gtgaagtgct gtctgacaga 961 gaattacatc tttcatggga agttggtaaa cctagaccac cacttaaccg aaattatgtc 1021 tttactggtt atcgtgtaac taaaaacagt aaagtacaaa taggagagta cacctttgaa 1081 aaaggtgact atggtgatgc tgttgtttac cgaggtacaa caacttacaa attaaatgtt 1141 ggtgattatt ttgtgctgac atcacataca gtaatgccat taagtgcacc tacactagtg 1201 ccacaagagc actatgttag aattactggc ttatacccaa cactcaatat ctcagatgag 1261 ttttctagca atgttgcaaa ttatcaaaag gttggtatgc aaaagtattc tacactccag 1321 ggaccacctg gtactggtaa gagtcatttt gctattggcc tagctctcta ctacccttct 1381 gctcgcatag tgtatacagc ttgctctcat gccgctgttg atgcactatg tgagaaggca 1441 ttaaaatatt tgcctataga taaatgtagt agaattatac ctgcacgtgc tcgtgtagag 1501 tgttttgata aattcaaagt gaattcaaca ttagaacagt atgtcttttg tactgtaaat 1561 gcattgcctg agacgacagc agatatagtt gtctttgatg aaatttcaat ggccacaaat 1621 tatgatttga gtgttgtcaa tgccagatta cgtgctaagc actatgtgta cattggcgac 1681 cctgctcaat tacctgcacc acgcacattg ctaactaagg gcacactaga accagaatat 1741 ttcaattcag tgtgtagact tatgaaaact ataggtccag acatgttcct cggaacttgt 1801 cggcgttgtc ctgctgaaat tgttgacact gtgagtgctt tggtttatga taataagctt 1861 aaagcacata aagacaaatc agctcaatgc tttaaaatgt tttataaggg tgttatcacg 1921 catgatgttt catctgcaat taacaggcca caaataggcg tggtaagaga attccttaca 1981 cgtaaccctg cttggagaaa agctgtcttt atttcacctt ataattcaca gaatgctgta 2041 gcctcaaaga ttttgggact accaactcaa actgttgatt catcacaggg ctcagaatat 2101 gactatgtca tattcactca aaccactgaa acagctcact cttgtaatgt aaacagattt 2161 aatgttgcta ttaccagagc aaaagtaggc atactttgca taatgtctga tagagacctt 2221 tatgacaagt tgcaatttac aagtcttgaa attccacgta ggaatgtggc aactttactc 2281 gtgccgagag gttcttggag ccatccgcag tttgaaaaat aaacaggccc cttttccttt 2341 gtcgatatca tgtaattagt tatgtcacgc ttacattcac gccctcctcc cacatccgct 2401 ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta ttttttttaa 2461 tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt tttctgtaca 2521 aagcggccgc cgaaccacta gtgcactgca gtacacggtt tccttgaaat ttttttgatt 2581 cggtaatctc cgaacagaag gaagaacgaa ggaaggagca cagacttaga ttggtatata 2641 tacgcatatg tagtgttgaa gaaacatgaa attgcccagt attcttaacc caactgcaca 2701 gaacaaaaac gtgcaggaaa cgaagataaa tcatgtcgaa agctacatat aaggaacgtg 2761 ctgctactca tcctagtcct gttgctgcca agctatttaa tatcatgcac gaaaagcaaa 2821 caaacttgtg tgcttcattg gatgttcgta ccaccaagga attactggag ttagttgaag 2881 cattaggtcc caaaatttgt ttactaaaaa cacatgtgga tatcttgact gatttttcca 2941 tggagggcac agttaagccg ctaaaggcat tatccgccaa gtacaatttt ttactcttcg 3001 aggacagaaa atttgctgac attggtaata cagtcaaatt gcagtactct gcgggtgtat 3061 acagaatagc agaatgggca gacattacga atgcacacgg tgtggtgggc ccaggtattg 3121 ttagcggttt gaagcaggcg gcagaagaag taacaaagga acctagaggc cttttgatgt 3181 tagcagaatt gtcatgcaag ggctccctat ctactggaga atatactaag ggtactgttg 3241 acattgcgaa gagcgacaaa gattttgtta tcggctttat tgctcaaaga gacatgggtg 3301 gaagagatga aggttacgat tggttgatta tgacacccgg tgtgggttta gatgacaagg 3361 gagatgcatt gggtcaacag tatagaaccg tggatgatgt ggtttctaca ggatctgaca 3421 ttattattgt tggaagagga ctatttgcaa agggaaggga tgctaaggta gagggtgaac 3481 gttacagaaa agcaggctgg gaagcatatt tgagaagatg cggccagcaa aactaaaaaa 3541 ctgtattata agtaaatgca tgtatactaa actcacaaat tagagcttca atttaattat 3601 atcagttatt acccgagtaa cgaagcatct gtgcttcatt ttgtagaaca aaaatgcaac 3661 gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc 3721 aacgcgaaag cgctatttta ccaacgaaga atctgtgctt catttttgta aaacaaaaat 3781 gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga 3841 aatgcaacgc gagagcgcta ttttaccaac aaagaatcta tacttctttt ttgttctaca 3901 aaaatgcatc ccgagagcgc tatttttcta acaaagcatc ttagattact ttttttctcc 3961 tttgtgcgct ctataatgca gtctcttgat aactttttgc actgtaggtc cgttaaggtt 4021 agaagaaggc tactttggtg tctattttct cttccataaa aaaagcctga ctccacttcc 4081 cgcgtttact gattactagc gaagctgcgg gtgcattttt tcaagataaa ggcatccccg 4141 attatattct ataccgatgt ggattgcgca tactttgtga acagaaagtg atagcgttga 4201 tgattcttca ttggtcagaa aattatgaac ggtttcttct attttgtctc tatatactac 4261 gtataggaaa tgtttacatt ttcgtattgt tttcgattca ctctatgaat agttcttact 4321 acaatttttt tgtctaaaga gtaatactag agataaacat aaaaaatgta gaggtcgagt 4381 ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt tatataggga tatagcacag 4441 agatatatag caaagagata cttttgagca atgtttgtgg aagcggtatt cgcaatattt 4501 tagtagctcg ttacagtccg gtgcgttttt ggttttttga aagtgcgtca tcagagcgct 4561 tttggttttc aaaagcgctc tgaagttcct atactttcta gctagagaat aggaacttcc 4621 cgagcggccg cgtgttacaa gcttgactcc agcgtaactg gactgcaatc aactcactgg 4681 ctcaccttca cgggtgggcc tttcttcggt agaaaatcaa aggatcttct tgagatcctt 4741 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt 4801 gtttgccgga tcaagagcta ccaactcttt ttccgaggta actggcttca gcagagcgca 4861 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt 4921 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga 4981 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc 5041 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact 5101 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga 5161 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggt 5221 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcatcgatt 5281 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cagaaaggcc 5341 cacccgaagg tgagccaggt gattacattt gggccctcat cagaggtttt caccgtcatc 5401 accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca 5461 gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg 5521 gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca cttaccaatg 5581 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg 5641 actccccgtc gtgtagataa ctacgatgcg ggagggctta ccatctggcc ccagtgctgc 5701 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc 5761 cgggagggcc gagcgcagaa gtgatcctgc aactttatcc gcctccatcc agtctattaa 5821 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc 5881 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg 5941 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc 6001 cttcggtcct ccgatcgttg ccagaagtaa gttggccgca gtgttatcac tcatggttat 6061 ggcagcactg cataattctc ttactgtcat gccatccgtg agatgctttt ctgtgactgg 6121 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc 6181 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg 6241 aaaacgttct tcggggcgta aactctcaag gatcttaccg ctgttgagat ccagttcgat 6301 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg 6361 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg 6421 ttgaatactc attttagctt ccttagctcc tgaaaatctc gataactcaa aaaatacgcc 6481 cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc gatcaagtca 6541 aaagcctccg gtcggaggct tttgactttc tgctatggag gtcaggtatg atttaaatgg 6601 tcagttaga //