LOCUS Exported 5307 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Yeast Expression plasmid for lSARS-CoV-2 eader protein. ACCESSION . VERSION . KEYWORDS pGBW-m4046686 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5307) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 5307) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5307 /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" primer_bind complement(1062..1080) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 1431..2234 /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 2317..3299 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 3377..3406 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 3428..4015 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 4027..4054 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(4078..4269) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(4270..5130) /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 4893..4912 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5131..5233) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 5234..5265 /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 gaatctttgg tccctggatt taacgagaaa acccacgtac agctatccct tcccgtgttg 541 caagttcgtg acgttctggt tcggggtttc ggtgattcag tcgaagaagt cttgagtgaa 601 gccaggcaac atttaaagga tggtacttgt ggcttagttg aagtcgaaaa gggtgtgttg 661 ccacaacttg agcaaccata cgttttcatt aagagatctg acgcaagaac agcgccacac 721 ggtcatgtta tggttgaatt ggtcgctgaa ttggagggta tacaatatgg cagatcgggt 781 gaaactttag gtgtattggt cccacacgta ggtgaaatcc ctgttgctta cagaaaagtt 841 ttattaagaa agaatggtaa caagggtgct ggtggacact cctacggagc cgatctgaag 901 tcttttgact tgggtgatga attgggtact gatccatatg aagacttcca agaaaattgg 961 aacaccaaac atagctctgg tgttacgcgt gaattgatga gagaacttaa tggtggttaa 1021 acaggcccct tttcctttgt cgatatcatg taattagtta tgtcacgctt acattcacgc 1081 cctcctccca catccgctct aaccgaaaag gaaggagtta gacaacctga agtctaggtc 1141 cctatttatt ttttttaata gttatgttag tattaagaac gttatttata tttcaaattt 1201 ttcttttttt tctgtacaaa gcggccgccg aaccactagt gcactgcagt acacggtttc 1261 cttgaaattt ttttgattcg gtaatctccg aacagaagga agaacgaagg aaggagcaca 1321 gacttagatt ggtatatata cgcatatgta gtgttgaaga aacatgaaat tgcccagtat 1381 tcttaaccca actgcacaga acaaaaacgt gcaggaaacg aagataaatc atgtcgaaag 1441 ctacatataa ggaacgtgct gctactcatc ctagtcctgt tgctgccaag ctatttaata 1501 tcatgcacga aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc accaaggaat 1561 tactggagtt agttgaagca ttaggtccca aaatttgttt actaaaaaca catgtggata 1621 tcttgactga tttttccatg gagggcacag ttaagccgct aaaggcatta tccgccaagt 1681 acaatttttt actcttcgag gacagaaaat ttgctgacat tggtaataca gtcaaattgc 1741 agtactctgc gggtgtatac agaatagcag aatgggcaga cattacgaat gcacacggtg 1801 tggtgggccc aggtattgtt agcggtttga agcaggcggc agaagaagta acaaaggaac 1861 ctagaggcct tttgatgtta gcagaattgt catgcaaggg ctccctatct actggagaat 1921 atactaaggg tactgttgac attgcgaaga gcgacaaaga ttttgttatc ggctttattg 1981 ctcaaagaga catgggtgga agagatgaag gttacgattg gttgattatg acacccggtg 2041 tgggtttaga tgacaaggga gatgcattgg gtcaacagta tagaaccgtg gatgatgtgg 2101 tttctacagg atctgacatt attattgttg gaagaggact atttgcaaag ggaagggatg 2161 ctaaggtaga gggtgaacgt tacagaaaag caggctggga agcatatttg agaagatgcg 2221 gccagcaaaa ctaaaaaact gtattataag taaatgcatg tatactaaac tcacaaatta 2281 gagcttcaat ttaattatat cagttattac ccgagtaacg aagcatctgt gcttcatttt 2341 gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt 2401 tttacagaac agaaatgcaa cgcgaaagcg ctattttacc aacgaagaat ctgtgcttca 2461 tttttgtaaa acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct 2521 gcatttttac agaacagaaa tgcaacgcga gagcgctatt ttaccaacaa agaatctata 2581 cttctttttt gttctacaaa aatgcatccc gagagcgcta tttttctaac aaagcatctt 2641 agattacttt ttttctcctt tgtgcgctct ataatgcagt ctcttgataa ctttttgcac 2701 tgtaggtccg ttaaggttag aagaaggcta ctttggtgtc tattttctct tccataaaaa 2761 aagcctgact ccacttcccg cgtttactga ttactagcga agctgcgggt gcattttttc 2821 aagataaagg catccccgat tatattctat accgatgtgg attgcgcata ctttgtgaac 2881 agaaagtgat agcgttgatg attcttcatt ggtcagaaaa ttatgaacgg tttcttctat 2941 tttgtctcta tatactacgt ataggaaatg tttacatttt cgtattgttt tcgattcact 3001 ctatgaatag ttcttactac aatttttttg tctaaagagt aatactagag ataaacataa 3061 aaaatgtaga ggtcgagttt agatgcaagt tcaaggagcg aaaggtggat gggtaggtta 3121 tatagggata tagcacagag atatatagca aagagatact tttgagcaat gtttgtggaa 3181 gcggtattcg caatatttta gtagctcgtt acagtccggt gcgtttttgg ttttttgaaa 3241 gtgcgtcatc agagcgcttt tggttttcaa aagcgctctg aagttcctat actttctagc 3301 tagagaatag gaacttcccg agcggccgcg tgttacaagc ttgactccag cgtaactgga 3361 ctgcaatcaa ctcactggct caccttcacg ggtgggcctt tcttcggtag aaaatcaaag 3421 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac 3481 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaggtaac 3541 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca 3601 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt 3661 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc 3721 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg 3781 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc 3841 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac 3901 gagggagctt ccaggggtaa acgcctggta tctttatagt cctgtcgggt ttcgccacct 3961 ctgacttgag catcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc 4021 cagcaacgca gaaaggccca cccgaaggtg agccaggtga ttacatttgg gccctcatca 4081 gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg 4141 gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc 4201 cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg 4261 tttggtcact taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg 4321 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatgcggg agggcttacc 4381 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc 4441 agcaataaac cagccagccg ggagggccga gcgcagaagt gatcctgcaa ctttatccgc 4501 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag 4561 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat 4621 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg 4681 caaaaaagcg gttagctcct tcggtcctcc gatcgttgcc agaagtaagt tggccgcagt 4741 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtgag 4801 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg 4861 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt 4921 aaaagtgctc atcattggaa aacgttcttc ggggcgtaaa ctctcaagga tcttaccgct 4981 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac 5041 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat 5101 aagggcgaca cggaaatgtt gaatactcat tttagcttcc ttagctcctg aaaatctcga 5161 taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt tggaacctct 5221 tacgtgccga tcaagtcaaa agcctccggt cggaggcttt tgactttctg ctatggaggt 5281 caggtatgat ttaaatggtc agttaga //