LOCUS Exported 5214 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Yeast Expression plasmid for SARS-CoV-2 ORF7a protein. ACCESSION . VERSION . KEYWORDS pGBW-m4046238 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5214) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 5214) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5214 /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 841..858 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" CDS 859..924 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" primer_bind complement(969..987) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 1338..2141 /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 2224..3206 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 3284..3313 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 3335..3922 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 3934..3961 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(3985..4176) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(4177..5037) /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 4800..4819 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5038..5140) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 5141..5172 /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 aaaattatcc tattcttagc attgataacc ctggctacgt gtgagcttta tcactaccag 541 gaatgcgtta gggggacaac tgtcttgttg aaggaacctt gttcatctgg aacttacgaa 601 ggcaattcgc catttcatcc attggctgat aacaagttcg ccttaacctg ttttagtact 661 caattcgctt tcgcttgtcc cgacggtgtg aaacacgttt atcaattgag agcgagatcc 721 gtaagcccaa agttatttat ccgtcaagaa gaagttcaag agttgtactc cccaattttt 781 ctaattgtcg ccgctatcgt tttcattacc ttgtgtttca cactgaagcg aaagactgaa 841 ctcgtgccga gaggttctga ttataaagat catgatggtg attataaaga tcatgatatt 901 gattataaag atgatgatga taaataaaca ggcccctttt cctttgtcga tatcatgtaa 961 ttagttatgt cacgcttaca ttcacgccct cctcccacat ccgctctaac cgaaaaggaa 1021 ggagttagac aacctgaagt ctaggtccct atttattttt tttaatagtt atgttagtat 1081 taagaacgtt atttatattt caaatttttc ttttttttct gtacaaagcg gccgccgaac 1141 cactagtgca ctgcagtaca cggtttcctt gaaatttttt tgattcggta atctccgaac 1201 agaaggaaga acgaaggaag gagcacagac ttagattggt atatatacgc atatgtagtg 1261 ttgaagaaac atgaaattgc ccagtattct taacccaact gcacagaaca aaaacgtgca 1321 ggaaacgaag ataaatcatg tcgaaagcta catataagga acgtgctgct actcatccta 1381 gtcctgttgc tgccaagcta tttaatatca tgcacgaaaa gcaaacaaac ttgtgtgctt 1441 cattggatgt tcgtaccacc aaggaattac tggagttagt tgaagcatta ggtcccaaaa 1501 tttgtttact aaaaacacat gtggatatct tgactgattt ttccatggag ggcacagtta 1561 agccgctaaa ggcattatcc gccaagtaca attttttact cttcgaggac agaaaatttg 1621 ctgacattgg taatacagtc aaattgcagt actctgcggg tgtatacaga atagcagaat 1681 gggcagacat tacgaatgca cacggtgtgg tgggcccagg tattgttagc ggtttgaagc 1741 aggcggcaga agaagtaaca aaggaaccta gaggcctttt gatgttagca gaattgtcat 1801 gcaagggctc cctatctact ggagaatata ctaagggtac tgttgacatt gcgaagagcg 1861 acaaagattt tgttatcggc tttattgctc aaagagacat gggtggaaga gatgaaggtt 1921 acgattggtt gattatgaca cccggtgtgg gtttagatga caagggagat gcattgggtc 1981 aacagtatag aaccgtggat gatgtggttt ctacaggatc tgacattatt attgttggaa 2041 gaggactatt tgcaaaggga agggatgcta aggtagaggg tgaacgttac agaaaagcag 2101 gctgggaagc atatttgaga agatgcggcc agcaaaacta aaaaactgta ttataagtaa 2161 atgcatgtat actaaactca caaattagag cttcaattta attatatcag ttattacccg 2221 agtaacgaag catctgtgct tcattttgta gaacaaaaat gcaacgcgag agcgctaatt 2281 tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc gaaagcgcta 2341 ttttaccaac gaagaatctg tgcttcattt ttgtaaaaca aaaatgcaac gcgagagcgc 2401 taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgagag 2461 cgctatttta ccaacaaaga atctatactt cttttttgtt ctacaaaaat gcatcccgag 2521 agcgctattt ttctaacaaa gcatcttaga ttactttttt tctcctttgt gcgctctata 2581 atgcagtctc ttgataactt tttgcactgt aggtccgtta aggttagaag aaggctactt 2641 tggtgtctat tttctcttcc ataaaaaaag cctgactcca cttcccgcgt ttactgatta 2701 ctagcgaagc tgcgggtgca ttttttcaag ataaaggcat ccccgattat attctatacc 2761 gatgtggatt gcgcatactt tgtgaacaga aagtgatagc gttgatgatt cttcattggt 2821 cagaaaatta tgaacggttt cttctatttt gtctctatat actacgtata ggaaatgttt 2881 acattttcgt attgttttcg attcactcta tgaatagttc ttactacaat ttttttgtct 2941 aaagagtaat actagagata aacataaaaa atgtagaggt cgagtttaga tgcaagttca 3001 aggagcgaaa ggtggatggg taggttatat agggatatag cacagagata tatagcaaag 3061 agatactttt gagcaatgtt tgtggaagcg gtattcgcaa tattttagta gctcgttaca 3121 gtccggtgcg tttttggttt tttgaaagtg cgtcatcaga gcgcttttgg ttttcaaaag 3181 cgctctgaag ttcctatact ttctagctag agaataggaa cttcccgagc ggccgcgtgt 3241 tacaagcttg actccagcgt aactggactg caatcaactc actggctcac cttcacgggt 3301 gggcctttct tcggtagaaa atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa 3361 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag 3421 agctaccaac tctttttccg aggtaactgg cttcagcaga gcgcagatac caaatactgt 3481 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata 3541 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac 3601 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg 3661 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg 3721 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag 3781 cggcagggtc ggaacaggag agcgcacgag ggagcttcca ggggtaaacg cctggtatct 3841 ttatagtcct gtcgggtttc gccacctctg acttgagcat cgatttttgt gatgctcgtc 3901 aggggggcgg agcctatgga aaaacgccag caacgcagaa aggcccaccc gaaggtgagc 3961 caggtgatta catttgggcc ctcatcagag gttttcaccg tcatcaccga aacgcgcgag 4021 gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc 4081 atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg 4141 ggccatgtta agggcggttt tttcctgttt ggtcacttac caatgcttaa tcagtgaggc 4201 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta 4261 gataactacg atgcgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga 4321 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggga gggccgagcg 4381 cagaagtgat cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc 4441 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat 4501 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag 4561 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat 4621 cgttgccaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa 4681 ttctcttact gtcatgccat ccgtgagatg cttttctgtg actggtgagt actcaaccaa 4741 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga 4801 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg 4861 gcgtaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc 4921 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg 4981 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatttt 5041 agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat 5101 ttcattatgg tgaaagttgg aacctcttac gtgccgatca agtcaaaagc ctccggtcgg 5161 aggcttttga ctttctgcta tggaggtcag gtatgattta aatggtcagt taga //