LOCUS Exported 4845 bp ds-DNA circular SYN 16-JUN-2021 DEFINITION Yeast Expression plasmid for SARS-CoV-2 nsp11. ACCESSION . VERSION . KEYWORDS pGBW-m4046521 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4845) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 4845) AUTHORS . TITLE Direct Submission JOURNAL Exported Jun 16, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4845 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..442 /gene="S. cerevisiae GAL1" /label=GAL1 promoter /note="inducible promoter, regulated by Gal4" protein_bind 1..118 /label=UAS /bound_moiety="Gal4" /note="upstream activating sequence mediating Gal4-dependent induction" primer_bind 413..436 /label=GAL1 /note="S. cerevisiae GAL1 promoter, forward primer" CDS 496..513 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" CDS 514..531 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(576..594) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 945..1748 /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 1831..2813 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 2891..2920 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 2942..3529 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 3541..3568 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(3592..3783) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(3784..4644) /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 4407..4426 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4645..4747) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 4748..4779 /label=tonB terminator /note="bidirectional E. coli tonB-P14 transcription terminator" ORIGIN 1 cggattagaa gccgccgagc gggcgacagc cctccgacgg aagactctcc tccgtgcgtc 61 ctcgtcttca ccggtcgcgt tcctgaaacg cagatgtgcc tcgcgccgca ctgctccgaa 121 caataaagat tctacaatac tagcttttat ggttatgaag aggaaaaatt ggcagtaacc 181 tggccccaca aaccttcaaa ttaacgaatc aaattaacaa ccataggatg ataatgcgat 241 tagtttttta gccttatttc tggggtaatt aatcagcgaa gcgatgattt ttgatctatt 301 aacagatata taaatggaaa agctgcataa ccactttaac taatactttc aacattttca 361 gtttgtatta cttcttattc aaatgtcata aaagtatcaa caaaaaattg ttaatatacc 421 tctatacttt aacgtcaagg agaaaaaact ataatgtccg ctgacgcaca atcattccta 481 aatgggtttg ctgtactcgt gccgagaggt tctcaccatc atcaccatca ttaaacaggc 541 cccttttcct ttgtcgatat catgtaatta gttatgtcac gcttacattc acgccctcct 601 cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta ggtccctatt 661 tatttttttt aatagttatg ttagtattaa gaacgttatt tatatttcaa atttttcttt 721 tttttctgta caaagcggcc gccgaaccac tagtgcactg cagtacacgg tttccttgaa 781 atttttttga ttcggtaatc tccgaacaga aggaagaacg aaggaaggag cacagactta 841 gattggtata tatacgcata tgtagtgttg aagaaacatg aaattgccca gtattcttaa 901 cccaactgca cagaacaaaa acgtgcagga aacgaagata aatcatgtcg aaagctacat 961 ataaggaacg tgctgctact catcctagtc ctgttgctgc caagctattt aatatcatgc 1021 acgaaaagca aacaaacttg tgtgcttcat tggatgttcg taccaccaag gaattactgg 1081 agttagttga agcattaggt cccaaaattt gtttactaaa aacacatgtg gatatcttga 1141 ctgatttttc catggagggc acagttaagc cgctaaaggc attatccgcc aagtacaatt 1201 ttttactctt cgaggacaga aaatttgctg acattggtaa tacagtcaaa ttgcagtact 1261 ctgcgggtgt atacagaata gcagaatggg cagacattac gaatgcacac ggtgtggtgg 1321 gcccaggtat tgttagcggt ttgaagcagg cggcagaaga agtaacaaag gaacctagag 1381 gccttttgat gttagcagaa ttgtcatgca agggctccct atctactgga gaatatacta 1441 agggtactgt tgacattgcg aagagcgaca aagattttgt tatcggcttt attgctcaaa 1501 gagacatggg tggaagagat gaaggttacg attggttgat tatgacaccc ggtgtgggtt 1561 tagatgacaa gggagatgca ttgggtcaac agtatagaac cgtggatgat gtggtttcta 1621 caggatctga cattattatt gttggaagag gactatttgc aaagggaagg gatgctaagg 1681 tagagggtga acgttacaga aaagcaggct gggaagcata tttgagaaga tgcggccagc 1741 aaaactaaaa aactgtatta taagtaaatg catgtatact aaactcacaa attagagctt 1801 caatttaatt atatcagtta ttacccgagt aacgaagcat ctgtgcttca ttttgtagaa 1861 caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca 1921 gaacagaaat gcaacgcgaa agcgctattt taccaacgaa gaatctgtgc ttcatttttg 1981 taaaacaaaa atgcaacgcg agagcgctaa tttttcaaac aaagaatctg agctgcattt 2041 ttacagaaca gaaatgcaac gcgagagcgc tattttacca acaaagaatc tatacttctt 2101 ttttgttcta caaaaatgca tcccgagagc gctatttttc taacaaagca tcttagatta 2161 ctttttttct cctttgtgcg ctctataatg cagtctcttg ataacttttt gcactgtagg 2221 tccgttaagg ttagaagaag gctactttgg tgtctatttt ctcttccata aaaaaagcct 2281 gactccactt cccgcgttta ctgattacta gcgaagctgc gggtgcattt tttcaagata 2341 aaggcatccc cgattatatt ctataccgat gtggattgcg catactttgt gaacagaaag 2401 tgatagcgtt gatgattctt cattggtcag aaaattatga acggtttctt ctattttgtc 2461 tctatatact acgtatagga aatgtttaca ttttcgtatt gttttcgatt cactctatga 2521 atagttctta ctacaatttt tttgtctaaa gagtaatact agagataaac ataaaaaatg 2581 tagaggtcga gtttagatgc aagttcaagg agcgaaaggt ggatgggtag gttatatagg 2641 gatatagcac agagatatat agcaaagaga tacttttgag caatgtttgt ggaagcggta 2701 ttcgcaatat tttagtagct cgttacagtc cggtgcgttt ttggtttttt gaaagtgcgt 2761 catcagagcg cttttggttt tcaaaagcgc tctgaagttc ctatactttc tagctagaga 2821 ataggaactt cccgagcggc cgcgtgttac aagcttgact ccagcgtaac tggactgcaa 2881 tcaactcact ggctcacctt cacgggtggg cctttcttcg gtagaaaatc aaaggatctt 2941 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac 3001 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgagg taactggctt 3061 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt 3121 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc 3181 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa 3241 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac 3301 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg 3361 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga 3421 gcttccaggg gtaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact 3481 tgagcatcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa 3541 cgcagaaagg cccacccgaa ggtgagccag gtgattacat ttgggccctc atcagaggtt 3601 ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg 3661 aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag 3721 cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt 3781 cacttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc 3841 catagttgcc tgactccccg tcgtgtagat aactacgatg cgggagggct taccatctgg 3901 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat 3961 aaaccagcca gccgggaggg ccgagcgcag aagtgatcct gcaactttat ccgcctccat 4021 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg 4081 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc 4141 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa 4201 agcggttagc tccttcggtc ctccgatcgt tgccagaagt aagttggccg cagtgttatc 4261 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg tgagatgctt 4321 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag 4381 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt 4441 gctcatcatt ggaaaacgtt cttcggggcg taaactctca aggatcttac cgctgttgag 4501 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac 4561 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc 4621 gacacggaaa tgttgaatac tcattttagc ttccttagct cctgaaaatc tcgataactc 4681 aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac ctcttacgtg 4741 ccgatcaagt caaaagcctc cggtcggagg cttttgactt tctgctatgg aggtcaggta 4801 tgatttaaat ggtcagttag agcggccgct tgctatattg aagta //