LOCUS Exported 5055 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Yeast Expression plasmid for SARS-CoV-2 nsp7. ACCESSION . VERSION . KEYWORDS pGBW-m4046165 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5055) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 5055) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5055 /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..765 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(810..828) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 1179..1982 /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 2065..3047 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 3125..3154 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 3176..3763 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 3775..3802 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(3826..4017) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(4018..4878) /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 4641..4660 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4879..4981) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 4982..5013 /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 aggttctcac catcatcacc atcattaaac aggccccttt 781 tcctttgtcg atatcatgta attagttatg tcacgcttac attcacgccc tcctcccaca 841 tccgctctaa ccgaaaagga aggagttaga caacctgaag tctaggtccc tatttatttt 901 ttttaatagt tatgttagta ttaagaacgt tatttatatt tcaaattttt cttttttttc 961 tgtacaaagc ggccgccgaa ccactagtgc actgcagtac acggtttcct tgaaattttt 1021 ttgattcggt aatctccgaa cagaaggaag aacgaaggaa ggagcacaga cttagattgg 1081 tatatatacg catatgtagt gttgaagaaa catgaaattg cccagtattc ttaacccaac 1141 tgcacagaac aaaaacgtgc aggaaacgaa gataaatcat gtcgaaagct acatataagg 1201 aacgtgctgc tactcatcct agtcctgttg ctgccaagct atttaatatc atgcacgaaa 1261 agcaaacaaa cttgtgtgct tcattggatg ttcgtaccac caaggaatta ctggagttag 1321 ttgaagcatt aggtcccaaa atttgtttac taaaaacaca tgtggatatc ttgactgatt 1381 tttccatgga gggcacagtt aagccgctaa aggcattatc cgccaagtac aattttttac 1441 tcttcgagga cagaaaattt gctgacattg gtaatacagt caaattgcag tactctgcgg 1501 gtgtatacag aatagcagaa tgggcagaca ttacgaatgc acacggtgtg gtgggcccag 1561 gtattgttag cggtttgaag caggcggcag aagaagtaac aaaggaacct agaggccttt 1621 tgatgttagc agaattgtca tgcaagggct ccctatctac tggagaatat actaagggta 1681 ctgttgacat tgcgaagagc gacaaagatt ttgttatcgg ctttattgct caaagagaca 1741 tgggtggaag agatgaaggt tacgattggt tgattatgac acccggtgtg ggtttagatg 1801 acaagggaga tgcattgggt caacagtata gaaccgtgga tgatgtggtt tctacaggat 1861 ctgacattat tattgttgga agaggactat ttgcaaaggg aagggatgct aaggtagagg 1921 gtgaacgtta cagaaaagca ggctgggaag catatttgag aagatgcggc cagcaaaact 1981 aaaaaactgt attataagta aatgcatgta tactaaactc acaaattaga gcttcaattt 2041 aattatatca gttattaccc gagtaacgaa gcatctgtgc ttcattttgt agaacaaaaa 2101 tgcaacgcga gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag 2161 aaatgcaacg cgaaagcgct attttaccaa cgaagaatct gtgcttcatt tttgtaaaac 2221 aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc atttttacag 2281 aacagaaatg caacgcgaga gcgctatttt accaacaaag aatctatact tcttttttgt 2341 tctacaaaaa tgcatcccga gagcgctatt tttctaacaa agcatcttag attacttttt 2401 ttctcctttg tgcgctctat aatgcagtct cttgataact ttttgcactg taggtccgtt 2461 aaggttagaa gaaggctact ttggtgtcta ttttctcttc cataaaaaaa gcctgactcc 2521 acttcccgcg tttactgatt actagcgaag ctgcgggtgc attttttcaa gataaaggca 2581 tccccgatta tattctatac cgatgtggat tgcgcatact ttgtgaacag aaagtgatag 2641 cgttgatgat tcttcattgg tcagaaaatt atgaacggtt tcttctattt tgtctctata 2701 tactacgtat aggaaatgtt tacattttcg tattgttttc gattcactct atgaatagtt 2761 cttactacaa tttttttgtc taaagagtaa tactagagat aaacataaaa aatgtagagg 2821 tcgagtttag atgcaagttc aaggagcgaa aggtggatgg gtaggttata tagggatata 2881 gcacagagat atatagcaaa gagatacttt tgagcaatgt ttgtggaagc ggtattcgca 2941 atattttagt agctcgttac agtccggtgc gtttttggtt ttttgaaagt gcgtcatcag 3001 agcgcttttg gttttcaaaa gcgctctgaa gttcctatac tttctagcta gagaatagga 3061 acttcccgag cggccgcgtg ttacaagctt gactccagcg taactggact gcaatcaact 3121 cactggctca ccttcacggg tgggcctttc ttcggtagaa aatcaaagga tcttcttgag 3181 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg 3241 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaggtaactg gcttcagcag 3301 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa 3361 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag 3421 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca 3481 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac 3541 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa 3601 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc 3661 aggggtaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagca 3721 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcaga 3781 aaggcccacc cgaaggtgag ccaggtgatt acatttgggc cctcatcaga ggttttcacc 3841 gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga 3901 ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa 3961 tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactta 4021 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt 4081 tgcctgactc cccgtcgtgt agataactac gatgcgggag ggcttaccat ctggccccag 4141 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca 4201 gccagccggg agggccgagc gcagaagtga tcctgcaact ttatccgcct ccatccagtc 4261 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt 4321 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag 4381 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt 4441 tagctccttc ggtcctccga tcgttgccag aagtaagttg gccgcagtgt tatcactcat 4501 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtgagat gcttttctgt 4561 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc 4621 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat 4681 cattggaaaa cgttcttcgg ggcgtaaact ctcaaggatc ttaccgctgt tgagatccag 4741 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt 4801 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg 4861 gaaatgttga atactcattt tagcttcctt agctcctgaa aatctcgata actcaaaaaa 4921 tacgcccggt agtgatctta tttcattatg gtgaaagttg gaacctctta cgtgccgatc 4981 aagtcaaaag cctccggtcg gaggcttttg actttctgct atggaggtca ggtatgattt 5041 aaatggtcag ttaga //