LOCUS       sequence-297570-        4992 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast expression plasmid for SARS-CoV-2 E (envelope).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4992)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 4992)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGBW-m4046705
FEATURES             Location/Qualifiers
     source          1..4992
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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(747..765)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1116..1919
                     /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      2002..2984
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3062..3091
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3113..3700
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3712..3739)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3763..3954)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(3955..4746,4747..4815))
                     /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     4578..4597
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4816..4918)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(4919..4950)
                     /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 tactcattcg tttcggaaga gacaggtacg ttaatagtta atagcgtact tctttttctt
      541 gctttcgtgg tattcttgct agttacacta gccatcctta ctgcgcttcg attgtgtgcg
      601 tactgctgca atattgttaa cgtgagtctt gtaaaacctt ctttttacgt ttactctcgt
      661 gttaaaaatc tgaattcttc tagagttcct gatcttctgg tctaaacagg ccccttttcc
      721 tttgtcgata tcatgtaatt agttatgtca cgcttacatt cacgccctcc tcccacatcc
      781 gctctaaccg aaaaggaagg agttagacaa cctgaagtct aggtccctat ttattttttt
      841 taatagttat gttagtatta agaacgttat ttatatttca aatttttctt ttttttctgt
      901 acaaagcggc cgccgaacca ctagtgcact gcagtacacg gtttccttga aatttttttg
      961 attcggtaat ctccgaacag aaggaagaac gaaggaagga gcacagactt agattggtat
     1021 atatacgcat atgtagtgtt gaagaaacat gaaattgccc agtattctta acccaactgc
     1081 acagaacaaa aacgtgcagg aaacgaagat aaatcatgtc gaaagctaca tataaggaac
     1141 gtgctgctac tcatcctagt cctgttgctg ccaagctatt taatatcatg cacgaaaagc
     1201 aaacaaactt gtgtgcttca ttggatgttc gtaccaccaa ggaattactg gagttagttg
     1261 aagcattagg tcccaaaatt tgtttactaa aaacacatgt ggatatcttg actgattttt
     1321 ccatggaggg cacagttaag ccgctaaagg cattatccgc caagtacaat tttttactct
     1381 tcgaggacag aaaatttgct gacattggta atacagtcaa attgcagtac tctgcgggtg
     1441 tatacagaat agcagaatgg gcagacatta cgaatgcaca cggtgtggtg ggcccaggta
     1501 ttgttagcgg tttgaagcag gcggcagaag aagtaacaaa ggaacctaga ggccttttga
     1561 tgttagcaga attgtcatgc aagggctccc tatctactgg agaatatact aagggtactg
     1621 ttgacattgc gaagagcgac aaagattttg ttatcggctt tattgctcaa agagacatgg
     1681 gtggaagaga tgaaggttac gattggttga ttatgacacc cggtgtgggt ttagatgaca
     1741 agggagatgc attgggtcaa cagtatagaa ccgtggatga tgtggtttct acaggatctg
     1801 acattattat tgttggaaga ggactatttg caaagggaag ggatgctaag gtagagggtg
     1861 aacgttacag aaaagcaggc tgggaagcat atttgagaag atgcggccag caaaactaaa
     1921 aaactgtatt ataagtaaat gcatgtatac taaactcaca aattagagct tcaatttaat
     1981 tatatcagtt attacccgag taacgaagca tctgtgcttc attttgtaga acaaaaatgc
     2041 aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa
     2101 tgcaacgcga aagcgctatt ttaccaacga agaatctgtg cttcattttt gtaaaacaaa
     2161 aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac
     2221 agaaatgcaa cgcgagagcg ctattttacc aacaaagaat ctatacttct tttttgttct
     2281 acaaaaatgc atcccgagag cgctattttt ctaacaaagc atcttagatt actttttttc
     2341 tcctttgtgc gctctataat gcagtctctt gataactttt tgcactgtag gtccgttaag
     2401 gttagaagaa ggctactttg gtgtctattt tctcttccat aaaaaaagcc tgactccact
     2461 tcccgcgttt actgattact agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc
     2521 ccgattatat tctataccga tgtggattgc gcatactttg tgaacagaaa gtgatagcgt
     2581 tgatgattct tcattggtca gaaaattatg aacggtttct tctattttgt ctctatatac
     2641 tacgtatagg aaatgtttac attttcgtat tgttttcgat tcactctatg aatagttctt
     2701 actacaattt ttttgtctaa agagtaatac tagagataaa cataaaaaat gtagaggtcg
     2761 agtttagatg caagttcaag gagcgaaagg tggatgggta ggttatatag ggatatagca
     2821 cagagatata tagcaaagag atacttttga gcaatgtttg tggaagcggt attcgcaata
     2881 ttttagtagc tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg tcatcagagc
     2941 gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt ctagctagag aataggaact
     3001 tcccgagcgg ccgcgtgtta caagcttgac tccagcgtaa ctggactgca atcaactcac
     3061 tggctcacct tcacgggtgg gcctttcttc ggtagaaaat caaaggatct tcttgagatc
     3121 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     3181 tttgtttgcc ggatcaagag ctaccaactc tttttccgag gtaactggct tcagcagagc
     3241 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     3301 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     3361 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     3421 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     3481 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     3541 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     3601 ggtaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcatcg
     3661 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcagaaag
     3721 gcccacccga aggtgagcca ggtgattaca tttgggccct catcagaggt tttcaccgtc
     3781 atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc
     3841 acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt
     3901 ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcacttacca
     3961 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4021 ctgactcccc gtcgtgtaga taactacgat gcgggagggc ttaccatctg gccccagtgc
     4081 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4141 agccgggagg gccgagcgca gaagtgatcc tgcaacttta tccgcctcca tccagtctat
     4201 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4261 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4321 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     4381 ctccttcggt cctccgatcg ttgccagaag taagttggcc gcagtgttat cactcatggt
     4441 tatggcagca ctgcataatt ctcttactgt catgccatcc gtgagatgct tttctgtgac
     4501 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     4561 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     4621 tggaaaacgt tcttcggggc gtaaactctc aaggatctta ccgctgttga gatccagttc
     4681 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     4741 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     4801 atgttgaata ctcattttag cttccttagc tcctgaaaat ctcgataact caaaaaatac
     4861 gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt gccgatcaag
     4921 tcaaaagcct ccggtcggag gcttttgact ttctgctatg gaggtcaggt atgatttaaa
     4981 tggtcagtta ga
//
