LOCUS       sequence-298032-        6024 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 E (envelope).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6024)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6024)
  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-m4046275
FEATURES             Location/Qualifiers
     source          1..6024
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             619..636
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1779..1797)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2148..2951
                     /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      3034..4016
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4094..4123
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4145..4732
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4744..4771)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4795..4986)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4987..5778,5779..5847))
                     /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     5610..5629
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5848..5950)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5951..5982)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgccc cacacaccat agcttcaaaa tgtttctact ccttttttac tcttccagat
       61 tttctcggac tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag catactaaat
      121 ttcccctctt tcttcctcta gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa
      181 aaaagagacc gcctcgtttc tttttcttcg tcttgaaaaw tttttttttt tgattttttt
      241 ctctttcgat gacctcccat tgatatttaa gttaataaac ggtcttcaat ttctcaagtt
      301 tcagtttcat ttttcttgtt ctattacaac tttttttact tcttgctcat tagaaagaaa
      361 gcatagcaat ctaatctaag ttttaattac aaaatgtatt cttttgttag cgaggaaacg
      421 ggaactctta tagtaaactc ggtgctatta ttcctggctt tcgtcgtttt tctcttggtt
      481 accttagcaa ttttgactgc cttgaggttg tgtgcgtact gctgtaatat cgtcaacgtt
      541 tcattggtca agccttcttt ctacgtttac tccagagtca aaaatttgaa ctcctctcgc
      601 gtcccagatt tattggttct cgtgccgaga ggttctaaaa tcgaggaagg taagctagtc
      661 atttggataa atggagataa gggttacaac ggtttggcag aagtaggcaa gaaatttgaa
      721 aaggacacag gtatcaaggt gacggttgaa catccagaca agttagaaga aaaattccct
      781 caagttgctg ctactggtga tggtcccgac attattttct gggcccacga tcgttttggt
      841 ggatatgctc agtcaggttt gctggctgag atcaccccag acaaggcctt ccaagataaa
      901 ttgtacccat tcacttggga tgctgtcaga tacaacggta agcttatagc atatccaatt
      961 gctgttgaag ccttatcgtt gatctacaat aaggacttgt tacctaaccc accaaagaca
     1021 tgggaagaaa ttccagcttt ggataaagag ctaaaggcca agggaaaaag cgctttgatg
     1081 tttaacttac aagaacctta tttcacctgg ccattaatcg ctgctgacgg tggttacgca
     1141 tttaaatacg aaaacggtaa gtatgacatt aaggatgtcg gcgttgataa tgctggtgca
     1201 aaggctggtt tgaccttctt ggttgattta ataaagaata aacacatgaa cgctgacact
     1261 gactactcta tcgccgaagc cgctttcaac aagggtgaaa ctgctatgac cataaatggt
     1321 ccatgggcct ggagtaacat tgatacttct aaagtaaact atggtgtcac tgttttgcct
     1381 acattcaaag gccaaccatc caagcccttt gtcggtgttc tatccgctgg catcaatgct
     1441 gcttctccaa acaaggagct agctaaagaa tttttggaaa attacttgtt aactgacgaa
     1501 ggtttggagg ccgttaacaa ggacaagcca ttgggtgctg tcgctttgaa atcttacgaa
     1561 gaagaattgg ctaaggatcc cagaattgcc gctaccatgg agaacgccca aaagggtgaa
     1621 attatgccaa acatcccaca aatgtcagcc ttctggtacg cagttaggac tgccgtgatt
     1681 aatgctgctt ccggtagaca aaccgtcgac gaagcattaa aagatgctca aacgtaaaca
     1741 ggcccctttt cctttgtcga tatcatgtaa ttagttatgt cacgcttaca ttcacgccct
     1801 cctcccacat ccgctctaac cgaaaaggaa ggagttagac aacctgaagt ctaggtccct
     1861 atttattttt tttaatagtt atgttagtat taagaacgtt atttatattt caaatttttc
     1921 ttttttttct gtacaaagcg gccgccgaac cactagtgca ctgcagtaca cggtttcctt
     1981 gaaatttttt tgattcggta atctccgaac agaaggaaga acgaaggaag gagcacagac
     2041 ttagattggt atatatacgc atatgtagtg ttgaagaaac atgaaattgc ccagtattct
     2101 taacccaact gcacagaaca aaaacgtgca ggaaacgaag ataaatcatg tcgaaagcta
     2161 catataagga acgtgctgct actcatccta gtcctgttgc tgccaagcta tttaatatca
     2221 tgcacgaaaa gcaaacaaac ttgtgtgctt cattggatgt tcgtaccacc aaggaattac
     2281 tggagttagt tgaagcatta ggtcccaaaa tttgtttact aaaaacacat gtggatatct
     2341 tgactgattt ttccatggag ggcacagtta agccgctaaa ggcattatcc gccaagtaca
     2401 attttttact cttcgaggac agaaaatttg ctgacattgg taatacagtc aaattgcagt
     2461 actctgcggg tgtatacaga atagcagaat gggcagacat tacgaatgca cacggtgtgg
     2521 tgggcccagg tattgttagc ggtttgaagc aggcggcaga agaagtaaca aaggaaccta
     2581 gaggcctttt gatgttagca gaattgtcat gcaagggctc cctatctact ggagaatata
     2641 ctaagggtac tgttgacatt gcgaagagcg acaaagattt tgttatcggc tttattgctc
     2701 aaagagacat gggtggaaga gatgaaggtt acgattggtt gattatgaca cccggtgtgg
     2761 gtttagatga caagggagat gcattgggtc aacagtatag aaccgtggat gatgtggttt
     2821 ctacaggatc tgacattatt attgttggaa gaggactatt tgcaaaggga agggatgcta
     2881 aggtagaggg tgaacgttac agaaaagcag gctgggaagc atatttgaga agatgcggcc
     2941 agcaaaacta aaaaactgta ttataagtaa atgcatgtat actaaactca caaattagag
     3001 cttcaattta attatatcag ttattacccg agtaacgaag catctgtgct tcattttgta
     3061 gaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt
     3121 acagaacaga aatgcaacgc gaaagcgcta ttttaccaac gaagaatctg tgcttcattt
     3181 ttgtaaaaca aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca
     3241 tttttacaga acagaaatgc aacgcgagag cgctatttta ccaacaaaga atctatactt
     3301 cttttttgtt ctacaaaaat gcatcccgag agcgctattt ttctaacaaa gcatcttaga
     3361 ttactttttt tctcctttgt gcgctctata atgcagtctc ttgataactt tttgcactgt
     3421 aggtccgtta aggttagaag aaggctactt tggtgtctat tttctcttcc ataaaaaaag
     3481 cctgactcca cttcccgcgt ttactgatta ctagcgaagc tgcgggtgca ttttttcaag
     3541 ataaaggcat ccccgattat attctatacc gatgtggatt gcgcatactt tgtgaacaga
     3601 aagtgatagc gttgatgatt cttcattggt cagaaaatta tgaacggttt cttctatttt
     3661 gtctctatat actacgtata ggaaatgttt acattttcgt attgttttcg attcactcta
     3721 tgaatagttc ttactacaat ttttttgtct aaagagtaat actagagata aacataaaaa
     3781 atgtagaggt cgagtttaga tgcaagttca aggagcgaaa ggtggatggg taggttatat
     3841 agggatatag cacagagata tatagcaaag agatactttt gagcaatgtt tgtggaagcg
     3901 gtattcgcaa tattttagta gctcgttaca gtccggtgcg tttttggttt tttgaaagtg
     3961 cgtcatcaga gcgcttttgg ttttcaaaag cgctctgaag ttcctatact ttctagctag
     4021 agaataggaa cttcccgagc ggccgcgtgt tacaagcttg actccagcgt aactggactg
     4081 caatcaactc actggctcac cttcacgggt gggcctttct tcggtagaaa atcaaaggat
     4141 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     4201 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aggtaactgg
     4261 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     4321 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4381 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4441 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     4501 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     4561 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     4621 ggagcttcca ggggtaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     4681 acttgagcat cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     4741 caacgcagaa aggcccaccc gaaggtgagc caggtgatta catttgggcc ctcatcagag
     4801 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
     4861 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
     4921 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
     4981 ggtcacttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5041 atccatagtt gcctgactcc ccgtcgtgta gataactacg atgcgggagg gcttaccatc
     5101 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5161 aataaaccag ccagccggga gggccgagcg cagaagtgat cctgcaactt tatccgcctc
     5221 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5281 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5341 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     5401 aaaagcggtt agctccttcg gtcctccgat cgttgccaga agtaagttgg ccgcagtgtt
     5461 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtgagatg
     5521 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5581 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5641 agtgctcatc attggaaaac gttcttcggg gcgtaaactc tcaaggatct taccgctgtt
     5701 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5761 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     5821 ggcgacacgg aaatgttgaa tactcatttt agcttcctta gctcctgaaa atctcgataa
     5881 ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg aacctcttac
     5941 gtgccgatca agtcaaaagc ctccggtcgg aggcttttga ctttctgcta tggaggtcag
     6001 gtatgattta aatggtcagt taga
//
