LOCUS Exported 5061 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Yeast Expression plasmid for SARS-CoV-2 nsp7. ACCESSION . VERSION . KEYWORDS pGBW-m4046222 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5061) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 5061) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5061 /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 730..747 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" CDS 748..771 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" primer_bind complement(816..834) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 1185..1988 /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 2071..3053 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 3131..3160 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 3182..3769 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 3781..3808 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(3832..4023) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(4024..4884) /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 4647..4666 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4885..4987) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 4988..5019 /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 tccaaaatgt cagacgtcaa gtgtacgtcg gtagttctcc tgagtgttct acaacagtta 541 cgtgtggaga gctcttccaa gctttgggca caatgcgtcc aattgcacaa tgatatttta 601 ttggccaaag atacaaccga agctttcgaa aagatggttt ctttgttgtc tgtcttgttg 661 tctatgcaag gtgctgttga cataaacaag ttgtgtgaag aaatgctaga taacagagct 721 actttacaac tcgtgccgag aggttcttgg agccatccgc agtttgaaaa ataaacaggc 781 cccttttcct ttgtcgatat catgtaatta gttatgtcac gcttacattc acgccctcct 841 cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta ggtccctatt 901 tatttttttt aatagttatg ttagtattaa gaacgttatt tatatttcaa atttttcttt 961 tttttctgta caaagcggcc gccgaaccac tagtgcactg cagtacacgg tttccttgaa 1021 atttttttga ttcggtaatc tccgaacaga aggaagaacg aaggaaggag cacagactta 1081 gattggtata tatacgcata tgtagtgttg aagaaacatg aaattgccca gtattcttaa 1141 cccaactgca cagaacaaaa acgtgcagga aacgaagata aatcatgtcg aaagctacat 1201 ataaggaacg tgctgctact catcctagtc ctgttgctgc caagctattt aatatcatgc 1261 acgaaaagca aacaaacttg tgtgcttcat tggatgttcg taccaccaag gaattactgg 1321 agttagttga agcattaggt cccaaaattt gtttactaaa aacacatgtg gatatcttga 1381 ctgatttttc catggagggc acagttaagc cgctaaaggc attatccgcc aagtacaatt 1441 ttttactctt cgaggacaga aaatttgctg acattggtaa tacagtcaaa ttgcagtact 1501 ctgcgggtgt atacagaata gcagaatggg cagacattac gaatgcacac ggtgtggtgg 1561 gcccaggtat tgttagcggt ttgaagcagg cggcagaaga agtaacaaag gaacctagag 1621 gccttttgat gttagcagaa ttgtcatgca agggctccct atctactgga gaatatacta 1681 agggtactgt tgacattgcg aagagcgaca aagattttgt tatcggcttt attgctcaaa 1741 gagacatggg tggaagagat gaaggttacg attggttgat tatgacaccc ggtgtgggtt 1801 tagatgacaa gggagatgca ttgggtcaac agtatagaac cgtggatgat gtggtttcta 1861 caggatctga cattattatt gttggaagag gactatttgc aaagggaagg gatgctaagg 1921 tagagggtga acgttacaga aaagcaggct gggaagcata tttgagaaga tgcggccagc 1981 aaaactaaaa aactgtatta taagtaaatg catgtatact aaactcacaa attagagctt 2041 caatttaatt atatcagtta ttacccgagt aacgaagcat ctgtgcttca ttttgtagaa 2101 caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca 2161 gaacagaaat gcaacgcgaa agcgctattt taccaacgaa gaatctgtgc ttcatttttg 2221 taaaacaaaa atgcaacgcg agagcgctaa tttttcaaac aaagaatctg agctgcattt 2281 ttacagaaca gaaatgcaac gcgagagcgc tattttacca acaaagaatc tatacttctt 2341 ttttgttcta caaaaatgca tcccgagagc gctatttttc taacaaagca tcttagatta 2401 ctttttttct cctttgtgcg ctctataatg cagtctcttg ataacttttt gcactgtagg 2461 tccgttaagg ttagaagaag gctactttgg tgtctatttt ctcttccata aaaaaagcct 2521 gactccactt cccgcgttta ctgattacta gcgaagctgc gggtgcattt tttcaagata 2581 aaggcatccc cgattatatt ctataccgat gtggattgcg catactttgt gaacagaaag 2641 tgatagcgtt gatgattctt cattggtcag aaaattatga acggtttctt ctattttgtc 2701 tctatatact acgtatagga aatgtttaca ttttcgtatt gttttcgatt cactctatga 2761 atagttctta ctacaatttt tttgtctaaa gagtaatact agagataaac ataaaaaatg 2821 tagaggtcga gtttagatgc aagttcaagg agcgaaaggt ggatgggtag gttatatagg 2881 gatatagcac agagatatat agcaaagaga tacttttgag caatgtttgt ggaagcggta 2941 ttcgcaatat tttagtagct cgttacagtc cggtgcgttt ttggtttttt gaaagtgcgt 3001 catcagagcg cttttggttt tcaaaagcgc tctgaagttc ctatactttc tagctagaga 3061 ataggaactt cccgagcggc cgcgtgttac aagcttgact ccagcgtaac tggactgcaa 3121 tcaactcact ggctcacctt cacgggtggg cctttcttcg gtagaaaatc aaaggatctt 3181 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac 3241 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgagg taactggctt 3301 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt 3361 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc 3421 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa 3481 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac 3541 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg 3601 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga 3661 gcttccaggg gtaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact 3721 tgagcatcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa 3781 cgcagaaagg cccacccgaa ggtgagccag gtgattacat ttgggccctc atcagaggtt 3841 ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg 3901 aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag 3961 cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt 4021 cacttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc 4081 catagttgcc tgactccccg tcgtgtagat aactacgatg cgggagggct taccatctgg 4141 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat 4201 aaaccagcca gccgggaggg ccgagcgcag aagtgatcct gcaactttat ccgcctccat 4261 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg 4321 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc 4381 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa 4441 agcggttagc tccttcggtc ctccgatcgt tgccagaagt aagttggccg cagtgttatc 4501 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg tgagatgctt 4561 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag 4621 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt 4681 gctcatcatt ggaaaacgtt cttcggggcg taaactctca aggatcttac cgctgttgag 4741 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac 4801 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc 4861 gacacggaaa tgttgaatac tcattttagc ttccttagct cctgaaaatc tcgataactc 4921 aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac ctcttacgtg 4981 ccgatcaagt caaaagcctc cggtcggagg cttttgactt tctgctatgg aggtcaggta 5041 tgatttaaat ggtcagttag a //