LOCUS Exported 5181 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Yeast Expression plasmid for SARS-CoV-2 nsp10. ACCESSION . VERSION . KEYWORDS pGBW-m4046636 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5181) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 5181) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5181 /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(936..954) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 1305..2108 /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 2191..3173 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 3251..3280 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 3302..3889 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 3901..3928 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(3952..4143) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(4144..5004) /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 4767..4786 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5005..5107) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 5108..5139 /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 ggtaatgcaa cagaagtgcc tgccaattca actgtattat ctttctgtgc ttttgctgta 541 gatgctgcta aagcttacaa agattatcta gctagtgggg gacaaccaat cactaattgt 601 gttaagatgt tgtgtacaca cactggtact ggtcaggcaa taacagttac accggaagcc 661 aatatggatc aagaatcctt tggtggtgca tcgtgttgtc tgtactgccg ttgccacata 721 gatcatccaa atcctaaagg attttgtgac ttaaaaggta agtatgtaca aatacctaca 781 acttgtgcta atgaccctgt gggttttaca cttaaaaaca cagtctgtac cgtctgcggt 841 atgtggaaag gttatggctg tagttgtgat caactccgcg aacccatgct ttaaacaggc 901 cccttttcct ttgtcgatat catgtaatta gttatgtcac gcttacattc acgccctcct 961 cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta ggtccctatt 1021 tatttttttt aatagttatg ttagtattaa gaacgttatt tatatttcaa atttttcttt 1081 tttttctgta caaagcggcc gccgaaccac tagtgcactg cagtacacgg tttccttgaa 1141 atttttttga ttcggtaatc tccgaacaga aggaagaacg aaggaaggag cacagactta 1201 gattggtata tatacgcata tgtagtgttg aagaaacatg aaattgccca gtattcttaa 1261 cccaactgca cagaacaaaa acgtgcagga aacgaagata aatcatgtcg aaagctacat 1321 ataaggaacg tgctgctact catcctagtc ctgttgctgc caagctattt aatatcatgc 1381 acgaaaagca aacaaacttg tgtgcttcat tggatgttcg taccaccaag gaattactgg 1441 agttagttga agcattaggt cccaaaattt gtttactaaa aacacatgtg gatatcttga 1501 ctgatttttc catggagggc acagttaagc cgctaaaggc attatccgcc aagtacaatt 1561 ttttactctt cgaggacaga aaatttgctg acattggtaa tacagtcaaa ttgcagtact 1621 ctgcgggtgt atacagaata gcagaatggg cagacattac gaatgcacac ggtgtggtgg 1681 gcccaggtat tgttagcggt ttgaagcagg cggcagaaga agtaacaaag gaacctagag 1741 gccttttgat gttagcagaa ttgtcatgca agggctccct atctactgga gaatatacta 1801 agggtactgt tgacattgcg aagagcgaca aagattttgt tatcggcttt attgctcaaa 1861 gagacatggg tggaagagat gaaggttacg attggttgat tatgacaccc ggtgtgggtt 1921 tagatgacaa gggagatgca ttgggtcaac agtatagaac cgtggatgat gtggtttcta 1981 caggatctga cattattatt gttggaagag gactatttgc aaagggaagg gatgctaagg 2041 tagagggtga acgttacaga aaagcaggct gggaagcata tttgagaaga tgcggccagc 2101 aaaactaaaa aactgtatta taagtaaatg catgtatact aaactcacaa attagagctt 2161 caatttaatt atatcagtta ttacccgagt aacgaagcat ctgtgcttca ttttgtagaa 2221 caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca 2281 gaacagaaat gcaacgcgaa agcgctattt taccaacgaa gaatctgtgc ttcatttttg 2341 taaaacaaaa atgcaacgcg agagcgctaa tttttcaaac aaagaatctg agctgcattt 2401 ttacagaaca gaaatgcaac gcgagagcgc tattttacca acaaagaatc tatacttctt 2461 ttttgttcta caaaaatgca tcccgagagc gctatttttc taacaaagca tcttagatta 2521 ctttttttct cctttgtgcg ctctataatg cagtctcttg ataacttttt gcactgtagg 2581 tccgttaagg ttagaagaag gctactttgg tgtctatttt ctcttccata aaaaaagcct 2641 gactccactt cccgcgttta ctgattacta gcgaagctgc gggtgcattt tttcaagata 2701 aaggcatccc cgattatatt ctataccgat gtggattgcg catactttgt gaacagaaag 2761 tgatagcgtt gatgattctt cattggtcag aaaattatga acggtttctt ctattttgtc 2821 tctatatact acgtatagga aatgtttaca ttttcgtatt gttttcgatt cactctatga 2881 atagttctta ctacaatttt tttgtctaaa gagtaatact agagataaac ataaaaaatg 2941 tagaggtcga gtttagatgc aagttcaagg agcgaaaggt ggatgggtag gttatatagg 3001 gatatagcac agagatatat agcaaagaga tacttttgag caatgtttgt ggaagcggta 3061 ttcgcaatat tttagtagct cgttacagtc cggtgcgttt ttggtttttt gaaagtgcgt 3121 catcagagcg cttttggttt tcaaaagcgc tctgaagttc ctatactttc tagctagaga 3181 ataggaactt cccgagcggc cgcgtgttac aagcttgact ccagcgtaac tggactgcaa 3241 tcaactcact ggctcacctt cacgggtggg cctttcttcg gtagaaaatc aaaggatctt 3301 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac 3361 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgagg taactggctt 3421 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt 3481 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc 3541 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa 3601 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac 3661 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg 3721 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga 3781 gcttccaggg gtaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact 3841 tgagcatcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa 3901 cgcagaaagg cccacccgaa ggtgagccag gtgattacat ttgggccctc atcagaggtt 3961 ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg 4021 aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag 4081 cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt 4141 cacttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc 4201 catagttgcc tgactccccg tcgtgtagat aactacgatg cgggagggct taccatctgg 4261 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat 4321 aaaccagcca gccgggaggg ccgagcgcag aagtgatcct gcaactttat ccgcctccat 4381 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg 4441 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc 4501 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa 4561 agcggttagc tccttcggtc ctccgatcgt tgccagaagt aagttggccg cagtgttatc 4621 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg tgagatgctt 4681 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag 4741 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt 4801 gctcatcatt ggaaaacgtt cttcggggcg taaactctca aggatcttac cgctgttgag 4861 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac 4921 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc 4981 gacacggaaa tgttgaatac tcattttagc ttccttagct cctgaaaatc tcgataactc 5041 aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac ctcttacgtg 5101 ccgatcaagt caaaagcctc cggtcggagg cttttgactt tctgctatgg aggtcaggta 5161 tgatttaaat ggtcagttag a //