LOCUS Exported 4950 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Yeast Expression plasmid for SARS-CoV-2 ORF6 protein. ACCESSION . VERSION . KEYWORDS pGBW-m4046706 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4950) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 4950) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4950 /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(705..723) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 1074..1877 /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 1960..2942 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 3020..3049 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 3071..3658 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 3670..3697 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(3721..3912) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(3913..4773) /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 4536..4555 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4774..4876) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 4877..4908 /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 tttcacctgg tcgacttcca agttacaatc gcagagattt tacttataat catgaggact 541 ttcaaggtgt ctatttggaa tttggattac attatcaact tgatcattaa aaacttgtca 601 aagagtttga ccgaaaataa gtattcccag ttagatgaag aacaacctat ggaaatagac 661 taaacaggcc ccttttcctt tgtcgatatc atgtaattag ttatgtcacg cttacattca 721 cgccctcctc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc tgaagtctag 781 gtccctattt atttttttta atagttatgt tagtattaag aacgttattt atatttcaaa 841 tttttctttt ttttctgtac aaagcggccg ccgaaccact agtgcactgc agtacacggt 901 ttccttgaaa tttttttgat tcggtaatct ccgaacagaa ggaagaacga aggaaggagc 961 acagacttag attggtatat atacgcatat gtagtgttga agaaacatga aattgcccag 1021 tattcttaac ccaactgcac agaacaaaaa cgtgcaggaa acgaagataa atcatgtcga 1081 aagctacata taaggaacgt gctgctactc atcctagtcc tgttgctgcc aagctattta 1141 atatcatgca cgaaaagcaa acaaacttgt gtgcttcatt ggatgttcgt accaccaagg 1201 aattactgga gttagttgaa gcattaggtc ccaaaatttg tttactaaaa acacatgtgg 1261 atatcttgac tgatttttcc atggagggca cagttaagcc gctaaaggca ttatccgcca 1321 agtacaattt tttactcttc gaggacagaa aatttgctga cattggtaat acagtcaaat 1381 tgcagtactc tgcgggtgta tacagaatag cagaatgggc agacattacg aatgcacacg 1441 gtgtggtggg cccaggtatt gttagcggtt tgaagcaggc ggcagaagaa gtaacaaagg 1501 aacctagagg ccttttgatg ttagcagaat tgtcatgcaa gggctcccta tctactggag 1561 aatatactaa gggtactgtt gacattgcga agagcgacaa agattttgtt atcggcttta 1621 ttgctcaaag agacatgggt ggaagagatg aaggttacga ttggttgatt atgacacccg 1681 gtgtgggttt agatgacaag ggagatgcat tgggtcaaca gtatagaacc gtggatgatg 1741 tggtttctac aggatctgac attattattg ttggaagagg actatttgca aagggaaggg 1801 atgctaaggt agagggtgaa cgttacagaa aagcaggctg ggaagcatat ttgagaagat 1861 gcggccagca aaactaaaaa actgtattat aagtaaatgc atgtatacta aactcacaaa 1921 ttagagcttc aatttaatta tatcagttat tacccgagta acgaagcatc tgtgcttcat 1981 tttgtagaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc 2041 atttttacag aacagaaatg caacgcgaaa gcgctatttt accaacgaag aatctgtgct 2101 tcatttttgt aaaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca aagaatctga 2161 gctgcatttt tacagaacag aaatgcaacg cgagagcgct attttaccaa caaagaatct 2221 atacttcttt tttgttctac aaaaatgcat cccgagagcg ctatttttct aacaaagcat 2281 cttagattac tttttttctc ctttgtgcgc tctataatgc agtctcttga taactttttg 2341 cactgtaggt ccgttaaggt tagaagaagg ctactttggt gtctattttc tcttccataa 2401 aaaaagcctg actccacttc ccgcgtttac tgattactag cgaagctgcg ggtgcatttt 2461 ttcaagataa aggcatcccc gattatattc tataccgatg tggattgcgc atactttgtg 2521 aacagaaagt gatagcgttg atgattcttc attggtcaga aaattatgaa cggtttcttc 2581 tattttgtct ctatatacta cgtataggaa atgtttacat tttcgtattg ttttcgattc 2641 actctatgaa tagttcttac tacaattttt ttgtctaaag agtaatacta gagataaaca 2701 taaaaaatgt agaggtcgag tttagatgca agttcaagga gcgaaaggtg gatgggtagg 2761 ttatataggg atatagcaca gagatatata gcaaagagat acttttgagc aatgtttgtg 2821 gaagcggtat tcgcaatatt ttagtagctc gttacagtcc ggtgcgtttt tggttttttg 2881 aaagtgcgtc atcagagcgc ttttggtttt caaaagcgct ctgaagttcc tatactttct 2941 agctagagaa taggaacttc ccgagcggcc gcgtgttaca agcttgactc cagcgtaact 3001 ggactgcaat caactcactg gctcaccttc acgggtgggc ctttcttcgg tagaaaatca 3061 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac 3121 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaggt 3181 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg 3241 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc 3301 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt 3361 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga 3421 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct 3481 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg 3541 cacgagggag cttccagggg taaacgcctg gtatctttat agtcctgtcg ggtttcgcca 3601 cctctgactt gagcatcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa 3661 cgccagcaac gcagaaaggc ccacccgaag gtgagccagg tgattacatt tgggccctca 3721 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc 3781 gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt 3841 ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc 3901 ctgtttggtc acttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 3961 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatgc gggagggctt 4021 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt 4081 atcagcaata aaccagccag ccgggagggc cgagcgcaga agtgatcctg caactttatc 4141 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa 4201 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg 4261 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt 4321 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gccagaagta agttggccgc 4381 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt 4441 gagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg 4501 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac 4561 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcgt aaactctcaa ggatcttacc 4621 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt 4681 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg 4741 aataagggcg acacggaaat gttgaatact cattttagct tccttagctc ctgaaaatct 4801 cgataactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa agttggaacc 4861 tcttacgtgc cgatcaagtc aaaagcctcc ggtcggaggc ttttgacttt ctgctatgga 4921 ggtcaggtat gatttaaatg gtcagttaga //