LOCUS Exported 5166 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Yeast Expression plasmid for SARS-CoV-2 ORF8 protein. ACCESSION . VERSION . KEYWORDS pGBW-m4046506 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5166) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 5166) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5166 /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..876 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(921..939) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 1290..2093 /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 2176..3158 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 3236..3265 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 3287..3874 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 3886..3913 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(3937..4128) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(4129..4989) /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 4752..4771 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4990..5092) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 5093..5124 /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 aaattcttgg tttttcttgg tatcataact acagtagccg ctttccacca ggaatgttcc 541 ttacaaagct gcacgcaaca tcaaccgtac gtcgttgatg acccatgtcc aatccacttc 601 tattcgaagt ggtacattag agttggagct aggaagtctg cacctttgat tgagctgtgt 661 gtcgatgaag ctggttcaaa gtctccaatt caatacatcg acattggtaa ttatactgtc 721 tcctgtttgc cctttaccat taactgccaa gaaccaaaat taggcagttt ggttgtccga 781 tgttctttct acgaagactt tttggaatat catgatgtgc gtgttgtcct agatttcatc 841 ctcgtgccga gaggttctca ccatcatcac catcattaaa caggcccctt ttcctttgtc 901 gatatcatgt aattagttat gtcacgctta cattcacgcc ctcctcccac atccgctcta 961 accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt tttttaatag 1021 ttatgttagt attaagaacg ttatttatat ttcaaatttt tctttttttt ctgtacaaag 1081 cggccgccga accactagtg cactgcagta cacggtttcc ttgaaatttt tttgattcgg 1141 taatctccga acagaaggaa gaacgaagga aggagcacag acttagattg gtatatatac 1201 gcatatgtag tgttgaagaa acatgaaatt gcccagtatt cttaacccaa ctgcacagaa 1261 caaaaacgtg caggaaacga agataaatca tgtcgaaagc tacatataag gaacgtgctg 1321 ctactcatcc tagtcctgtt gctgccaagc tatttaatat catgcacgaa aagcaaacaa 1381 acttgtgtgc ttcattggat gttcgtacca ccaaggaatt actggagtta gttgaagcat 1441 taggtcccaa aatttgttta ctaaaaacac atgtggatat cttgactgat ttttccatgg 1501 agggcacagt taagccgcta aaggcattat ccgccaagta caatttttta ctcttcgagg 1561 acagaaaatt tgctgacatt ggtaatacag tcaaattgca gtactctgcg ggtgtataca 1621 gaatagcaga atgggcagac attacgaatg cacacggtgt ggtgggccca ggtattgtta 1681 gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc tagaggcctt ttgatgttag 1741 cagaattgtc atgcaagggc tccctatcta ctggagaata tactaagggt actgttgaca 1801 ttgcgaagag cgacaaagat tttgttatcg gctttattgc tcaaagagac atgggtggaa 1861 gagatgaagg ttacgattgg ttgattatga cacccggtgt gggtttagat gacaagggag 1921 atgcattggg tcaacagtat agaaccgtgg atgatgtggt ttctacagga tctgacatta 1981 ttattgttgg aagaggacta tttgcaaagg gaagggatgc taaggtagag ggtgaacgtt 2041 acagaaaagc aggctgggaa gcatatttga gaagatgcgg ccagcaaaac taaaaaactg 2101 tattataagt aaatgcatgt atactaaact cacaaattag agcttcaatt taattatatc 2161 agttattacc cgagtaacga agcatctgtg cttcattttg tagaacaaaa atgcaacgcg 2221 agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac 2281 gcgaaagcgc tattttacca acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca 2341 acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat 2401 gcaacgcgag agcgctattt taccaacaaa gaatctatac ttcttttttg ttctacaaaa 2461 atgcatcccg agagcgctat ttttctaaca aagcatctta gattactttt tttctccttt 2521 gtgcgctcta taatgcagtc tcttgataac tttttgcact gtaggtccgt taaggttaga 2581 agaaggctac tttggtgtct attttctctt ccataaaaaa agcctgactc cacttcccgc 2641 gtttactgat tactagcgaa gctgcgggtg cattttttca agataaaggc atccccgatt 2701 atattctata ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata gcgttgatga 2761 ttcttcattg gtcagaaaat tatgaacggt ttcttctatt ttgtctctat atactacgta 2821 taggaaatgt ttacattttc gtattgtttt cgattcactc tatgaatagt tcttactaca 2881 atttttttgt ctaaagagta atactagaga taaacataaa aaatgtagag gtcgagttta 2941 gatgcaagtt caaggagcga aaggtggatg ggtaggttat atagggatat agcacagaga 3001 tatatagcaa agagatactt ttgagcaatg tttgtggaag cggtattcgc aatattttag 3061 tagctcgtta cagtccggtg cgtttttggt tttttgaaag tgcgtcatca gagcgctttt 3121 ggttttcaaa agcgctctga agttcctata ctttctagct agagaatagg aacttcccga 3181 gcggccgcgt gttacaagct tgactccagc gtaactggac tgcaatcaac tcactggctc 3241 accttcacgg gtgggccttt cttcggtaga aaatcaaagg atcttcttga gatccttttt 3301 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt 3361 tgccggatca agagctacca actctttttc cgaggtaact ggcttcagca gagcgcagat 3421 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc 3481 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa 3541 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg 3601 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag 3661 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag 3721 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc caggggtaaa 3781 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc atcgattttt 3841 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcag aaaggcccac 3901 ccgaaggtga gccaggtgat tacatttggg ccctcatcag aggttttcac cgtcatcacc 3961 gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat 4021 gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct 4081 tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactt accaatgctt 4141 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 4201 ccccgtcgtg tagataacta cgatgcggga gggcttacca tctggcccca gtgctgcaat 4261 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 4321 gagggccgag cgcagaagtg atcctgcaac tttatccgcc tccatccagt ctattaattg 4381 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 4441 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 4501 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 4561 cggtcctccg atcgttgcca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 4621 agcactgcat aattctctta ctgtcatgcc atccgtgaga tgcttttctg tgactggtga 4681 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 4741 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 4801 acgttcttcg gggcgtaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 4861 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 4921 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 4981 aatactcatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa atacgcccgg 5041 tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat caagtcaaaa 5101 gcctccggtc ggaggctttt gactttctgc tatggaggtc aggtatgatt taaatggtca 5161 gttaga //