LOCUS Exported 5130 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Yeast Expression plasmid for SARS-CoV-2 ORF8 protein. ACCESSION . VERSION . KEYWORDS pGBW-m4046707 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5130) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 5130) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5130 /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(885..903) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 1254..2057 /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 2140..3122 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 3200..3229 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 3251..3838 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 3850..3877 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(3901..4092) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(4093..4953) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPET?VKVKDAEDQLGARVGYI ELDLNSGKILESLRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLATIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGSLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 4716..4735 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4954..5056) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 5057..5088 /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 aaatttcttg ttttcttagg aatcatcaca actgtagctg catttcacca agaatgtagt 541 ttacagtcat gtactcaaca tcaaccatat gtagttgatg acccgtgtcc tattcacttc 601 tattctaaat ggtatattag agtaggagct agaaaatcag cacctttaat tgaattgtgc 661 gtggatgagg ctggttctaa atcacccatt cagtacatcg atatcggtaa ttatacagtt 721 tcctgtttac cttttacaat taattgccag gaacctaaat tgggtagtct tgtagtgcgt 781 tgttcgttct atgaagactt tttagagtat catgacgttc gtgttgtttt agatttcatc 841 taaacaggcc ccttttcctt tgtcgatatc atgtaattag ttatgtcacg cttacattca 901 cgccctcctc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc tgaagtctag 961 gtccctattt atttttttta atagttatgt tagtattaag aacgttattt atatttcaaa 1021 tttttctttt ttttctgtac aaagcggccg ccgaaccact agtgcactgc agtacacggt 1081 ttccttgaaa tttttttgat tcggtaatct ccgaacagaa ggaagaacga aggaaggagc 1141 acagacttag attggtatat atacgcatat gtagtgttga agaaacatga aattgcccag 1201 tattcttaac ccaactgcac agaacaaaaa cgtgcaggaa acgaagataa atcatgtcga 1261 aagctacata taaggaacgt gctgctactc atcctagtcc tgttgctgcc aagctattta 1321 atatcatgca cgaaaagcaa acaaacttgt gtgcttcatt ggatgttcgt accaccaagg 1381 aattactgga gttagttgaa gcattaggtc ccaaaatttg tttactaaaa acacatgtgg 1441 atatcttgac tgatttttcc atggagggca cagttaagcc gctaaaggca ttatccgcca 1501 agtacaattt tttactcttc gaggacagaa aatttgctga cattggtaat acagtcaaat 1561 tgcagtactc tgcgggtgta tacagaatag cagaatgggc agacattacg aatgcacacg 1621 gtgtggtggg cccaggtatt gttagcggtt tgaagcaggc ggcagaagaa gtaacaaagg 1681 aacctagagg ccttttgatg ttagcagaat tgtcatgcaa gggctcccta tctactggag 1741 aatatactaa gggtactgtt gacattgcga agagcgacaa agattttgtt atcggcttta 1801 ttgctcaaag agacatgggt ggaagagatg aaggttacga ttggttgatt atgacacccg 1861 gtgtgggttt agatgacaag ggagatgcat tgggtcaaca gtatagaacc gtggatgatg 1921 tggtttctac aggatctgac attattattg ttggaagagg actatttgca aagggaaggg 1981 atgctaaggt agagggtgaa cgttacagaa aagcaggctg ggaagcatat ttgagaagat 2041 gcggccagca aaactaaaaa actgtattat aagtaaatgc atgtatacta aactcacaaa 2101 ttagagcttc aatttaatta tatcagttat tacccgagta acgaagcatc tgtgcttcat 2161 tttgtagaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc 2221 atttttacag aacagaaatg caacgcgaaa gcgctatttt accaacgaag aatctgtgct 2281 tcatttttgt aaaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca aagaatctga 2341 gctgcatttt tacagaacag aaatgcaacg cgagagcgct attttaccaa caaagaatct 2401 atacttcttt tttgttctac aaaaatgcat cccgagagcg ctatttttct aacaaagcat 2461 cttagattac tttttttctc ctttgtgcgc tctataatgc agtctcttga taactttttg 2521 cactgtaggt ccgttaaggt tagaagaagg ctactttggt gtctattttc tcttccataa 2581 aaaaagcctg actccacttc ccgcgtttac tgattactag cgaagctgcg ggtgcatttt 2641 ttcaagataa aggcatcccc gattatattc tataccgatg tggattgcgc atactttgtg 2701 aacagaaagt gatagcgttg atgattcttc attggtcaga aaattatgaa cggtttcttc 2761 tattttgtct ctatatacta cgtataggaa atgtttacat tttcgtattg ttttcgattc 2821 actctatgaa tagttcttac tacaattttt ttgtctaaag agtaatacta gagataaaca 2881 taaaaaatgt agaggtcgag tttagatgca agttcaagga gcgaaaggtg gatgggtagg 2941 ttatataggg atatagcaca gagatatata gcaaagagat acttttgagc aatgtttgtg 3001 gaagcggtat tcgcaatatt ttagtagctc gttacagtcc ggtgcgtttt tggttttttg 3061 aaagtgcgtc atcagagcgc ttttggtttt caaaagcgct ctgaagttcc tatactttct 3121 agctagagaa taggaacttc ccgagcggcc gcgtgttaca agcttgactc cagcgtaact 3181 ggactgcaat caactcactg gctcaccttc acgggtgggc ctttcttcgg tagaaaatca 3241 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac 3301 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaggt 3361 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg 3421 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc 3481 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt 3541 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga 3601 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct 3661 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg 3721 cacgagggag cttccagggg taaacgcctg gtatctttat agtcctgtcg ggtttcgcca 3781 cctctgactt gagcatcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa 3841 cgccagcaac gcagaaaggc ccacccgaag gtgagccagg tgattacatt tgggccctca 3901 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc 3961 gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt 4021 ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc 4081 ctgtttggtc acttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 4141 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatgc gggagggctt 4201 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt 4261 atcagcaata aaccagccag ccgggagggc cgagcgcaga agtgatcctg caactttatc 4321 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa 4381 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg 4441 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt 4501 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gccagaagta agttggccgc 4561 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt 4621 gagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg 4681 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac 4741 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcgt aaactctcaa ggatcttacc 4801 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt 4861 tactttcacc nnngtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg 4921 aataagggcg acacggaaat gttgaatact cattttagct tccttagctc ctgaaaatct 4981 cgataactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa agttggaacc 5041 tcttacgtgc cgatcaagtc aaaagcctcc ggtcggaggc ttttgacttt ctgctatgga 5101 ggtcaggtat gatttaaatg gtcagttaga //