LOCUS       sequence-297569-        8622 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast expression plasmid for SARS-CoV-2 S (surface glycoprotein).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8622)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 8622)
  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-m4046406
FEATURES             Location/Qualifiers
     source          1..8622
                     /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"
     CDS             4297..4314
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             4315..4332
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4377..4395)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             4746..5549
                     /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      5632..6614
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      6692..6721
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      6743..7330
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(7342..7369)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(7393..7584)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(7585..8376,8377..8445))
                     /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     8208..8227
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8446..8548)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(8549..8580)
                     /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 tttgtttttc ttgttttatt gccactagtc tctagtcagt gtgttaatct tacaaccaga
      541 actcaattac cccctgcata cactaattct ttcacacgtg gtgtttatta ccctgacaaa
      601 gttttcagat cctcagtttt acattcaact caggacttgt tcttaccttt cttttccaat
      661 gttacttggt tccatgctat acatgtctct gggaccaatg gtactaagag gtttgataac
      721 cctgtcctac catttaatga tggtgtttat tttgcttcca ctgagaagtc taacataata
      781 agaggctgga tttttggtac tactttagat tcgaagaccc agtccctact tattgttaat
      841 aacgctacta atgttgttat taaagtctgt gaatttcaat tttgtaatga tccatttttg
      901 ggtgtttatt accacaaaaa caacaaaagt tggatggaaa gtgagttcag agtttattct
      961 agtgcgaata attgcacttt tgaatatgtc tctcagcctt ttcttatgga ccttgaagga
     1021 aaacagggta atttcaaaaa tcttagggaa tttgtgttta agaatattga tggttatttt
     1081 aaaatatatt ctaagcacac gcctattaat ttagtgcgtg atctccctca gggtttttcg
     1141 gctttagaac cattggtaga tttgccaata ggtattaaca tcactaggtt tcaaacttta
     1201 cttgctttac atagaagtta tttgactcct ggtgattctt cttcaggttg gacagctggt
     1261 gctgcagctt attatgtggg ttatcttcaa cctaggactt ttctattaaa atataatgaa
     1321 aatggaacca ttacagatgc tgtagactgt gcacttgacc ctctctcaga aacaaagtgt
     1381 acgttgaaat ccttcactgt agaaaaagga atctatcaaa cttctaactt tagagtccaa
     1441 ccaacagaat ctattgttag atttcctaat attacaaact tgtgcccttt tggtgaagtt
     1501 tttaacgcca ccagatttgc atctgtttat gcttggaaca ggaagagaat cagcaactgt
     1561 gttgctgatt attctgtcct atataattcc gcatcatttt ccacttttaa gtgttatgga
     1621 gtgtctccta ctaaattaaa tgatctctgc tttactaatg tctatgcaga ttcatttgta
     1681 attagaggtg atgaagtcag acaaatcgct ccagggcaaa ctggaaagat tgctgattat
     1741 aattataaat taccagatga ttttacaggc tgcgttatag cttggaattc taacaatctt
     1801 gattctaagg ttggtggtaa ttataattac ctgtatagat tgtttaggaa gtctaatctc
     1861 aaaccttttg agagagatat ttcaactgaa atctatcagg ccggtagcac accttgtaat
     1921 ggtgttgaag gttttaattg ttactttcct ttacaatcat atggtttcca acccactaat
     1981 ggtgttggtt accaaccata cagagtagta gtactttctt ttgaacttct acatgcacca
     2041 gcaactgttt gtggacctaa aaagtctact aatttggtta aaaacaaatg tgtcaatttc
     2101 aacttcaatg gtttaacagg cacaggtgtt cttactgagt ctaacaaaaa gtttctgcct
     2161 ttccaacaat ttggcagaga cattgctgac actactgatg ctgtccgtga tccacagaca
     2221 cttgagattc ttgacattac accatgttct tttggtggtg tcagtgttat aacaccagga
     2281 acaaatactt ctaaccaggt tgctgttctt tatcaggatg ttaactgcac agaagtccct
     2341 gttgctattc atgcagatca acttactcct acttggcgtg tttattctac aggttctaat
     2401 gtttttcaaa cacgtgcagg ctgtttaata ggggctgaac atgtcaacaa ctcatatgag
     2461 tgtgacatac ccattggtgc aggtatatgc gctagttatc agactcagac taattctcct
     2521 cggcgggcac gtagtgtagc tagtcaatcc atcattgcct acactatgtc acttggtgca
     2581 gaaaattcag ttgcttactc taataactct attgccatac ccacaaattt tactattagt
     2641 gttaccacag aaattctacc agtgtctatg accaagacat cagtagattg tacaatgtac
     2701 atttgtggtg attcaactga atgcagcaat cttttgttgc aatatggcag tttttgtaca
     2761 caattaaacc gtgctttaac tggaatagct gttgaacaag acaaaaacac ccaagaagtt
     2821 tttgcacaag tcaaacaaat ttacaaaaca ccaccaatta aagattttgg tggttttaat
     2881 ttttcacaaa tattaccaga tccatcaaaa ccaagcaaga ggtcatttat tgaagatcta
     2941 cttttcaaca aagtgacact tgcagatgct ggcttcatca aacaatatgg tgattgcctt
     3001 ggtgatattg ctgctagaga cctcatttgt gcacaaaagt ttaacggcct tactgttttg
     3061 ccacctttgc tcacagatga aatgattgct caatacactt ctgcactgtt agcgggtaca
     3121 atcacttctg gttggacctt tggtgcaggt gctgcattac aaataccatt tgctatgcaa
     3181 atggcttata ggtttaatgg tattggagtt acacagaatg ttctctatga gaaccaaaaa
     3241 ttgattgcca accaatttaa tagtgctatt ggcaaaattc aagactcact ttcttccaca
     3301 gcaagtgcac ttggaaaact tcaagatgtg gtcaaccaaa atgcacaagc tttaaacacg
     3361 cttgttaaac aacttagctc caattttggt gcaatttcaa gtgttttaaa tgatatcctt
     3421 tcacgtcttg acaaagttga ggctgaagtg caaattgata ggttgatcac aggcagactt
     3481 caaagtttgc agacatatgt gactcaacaa ttaattagag ctgcagaaat cagagcttct
     3541 gctaatcttg ctgctactaa aatgtcagag tgtgtacttg gacaatcaaa aagagttgat
     3601 ttttgtggaa agggctatca tcttatgtcc ttccctcagt cagcacctca tggtgtagtc
     3661 ttcttgcatg tgacttatgt ccctgcacaa gaaaagaact tcacaactgc tcctgccatt
     3721 tgtcatgatg gaaaagcaca ctttcctcgt gaaggtgtct ttgtttcaaa tggcacacac
     3781 tggtttgtaa cacaaaggaa tttttatgaa ccacaaatca ttactacaga caacacattt
     3841 gtgtctggta actgtgatgt tgtaatagga attgtcaaca acacagttta tgatcctttg
     3901 caacctgaat tagactcatt caaggaggag ttagataaat attttaagaa tcatacatca
     3961 ccagatgttg atttaggtga catctctggc attaatgctt cagttgtaaa cattcaaaaa
     4021 gaaattgacc gcctcaatga ggttgccaag aatttaaatg aatctctcat cgatctccaa
     4081 gaacttggaa agtatgagca gtatataaaa tggccatggt acatttggct aggttttata
     4141 gctggcttga ttgccatagt aatggtgaca attatgcttt gctgtatgac cagttgctgt
     4201 agttgtctca agggctgttg ttcttgtgga tcctgctgca aatttgatga agacgactct
     4261 gagccagtgc tcaaaggagt caaattacat tacacactcg tgccgagagg ttctcaccat
     4321 catcaccatc attaaacagg ccccttttcc tttgtcgata tcatgtaatt agttatgtca
     4381 cgcttacatt cacgccctcc tcccacatcc gctctaaccg aaaaggaagg agttagacaa
     4441 cctgaagtct aggtccctat ttattttttt taatagttat gttagtatta agaacgttat
     4501 ttatatttca aatttttctt ttttttctgt acaaagcggc cgccgaacca ctagtgcact
     4561 gcagtacacg gtttccttga aatttttttg attcggtaat ctccgaacag aaggaagaac
     4621 gaaggaagga gcacagactt agattggtat atatacgcat atgtagtgtt gaagaaacat
     4681 gaaattgccc agtattctta acccaactgc acagaacaaa aacgtgcagg aaacgaagat
     4741 aaatcatgtc gaaagctaca tataaggaac gtgctgctac tcatcctagt cctgttgctg
     4801 ccaagctatt taatatcatg cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc
     4861 gtaccaccaa ggaattactg gagttagttg aagcattagg tcccaaaatt tgtttactaa
     4921 aaacacatgt ggatatcttg actgattttt ccatggaggg cacagttaag ccgctaaagg
     4981 cattatccgc caagtacaat tttttactct tcgaggacag aaaatttgct gacattggta
     5041 atacagtcaa attgcagtac tctgcgggtg tatacagaat agcagaatgg gcagacatta
     5101 cgaatgcaca cggtgtggtg ggcccaggta ttgttagcgg tttgaagcag gcggcagaag
     5161 aagtaacaaa ggaacctaga ggccttttga tgttagcaga attgtcatgc aagggctccc
     5221 tatctactgg agaatatact aagggtactg ttgacattgc gaagagcgac aaagattttg
     5281 ttatcggctt tattgctcaa agagacatgg gtggaagaga tgaaggttac gattggttga
     5341 ttatgacacc cggtgtgggt ttagatgaca agggagatgc attgggtcaa cagtatagaa
     5401 ccgtggatga tgtggtttct acaggatctg acattattat tgttggaaga ggactatttg
     5461 caaagggaag ggatgctaag gtagagggtg aacgttacag aaaagcaggc tgggaagcat
     5521 atttgagaag atgcggccag caaaactaaa aaactgtatt ataagtaaat gcatgtatac
     5581 taaactcaca aattagagct tcaatttaat tatatcagtt attacccgag taacgaagca
     5641 tctgtgcttc attttgtaga acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag
     5701 aatctgagct gcatttttac agaacagaaa tgcaacgcga aagcgctatt ttaccaacga
     5761 agaatctgtg cttcattttt gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa
     5821 caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgagagcg ctattttacc
     5881 aacaaagaat ctatacttct tttttgttct acaaaaatgc atcccgagag cgctattttt
     5941 ctaacaaagc atcttagatt actttttttc tcctttgtgc gctctataat gcagtctctt
     6001 gataactttt tgcactgtag gtccgttaag gttagaagaa ggctactttg gtgtctattt
     6061 tctcttccat aaaaaaagcc tgactccact tcccgcgttt actgattact agcgaagctg
     6121 cgggtgcatt ttttcaagat aaaggcatcc ccgattatat tctataccga tgtggattgc
     6181 gcatactttg tgaacagaaa gtgatagcgt tgatgattct tcattggtca gaaaattatg
     6241 aacggtttct tctattttgt ctctatatac tacgtatagg aaatgtttac attttcgtat
     6301 tgttttcgat tcactctatg aatagttctt actacaattt ttttgtctaa agagtaatac
     6361 tagagataaa cataaaaaat gtagaggtcg agtttagatg caagttcaag gagcgaaagg
     6421 tggatgggta ggttatatag ggatatagca cagagatata tagcaaagag atacttttga
     6481 gcaatgtttg tggaagcggt attcgcaata ttttagtagc tcgttacagt ccggtgcgtt
     6541 tttggttttt tgaaagtgcg tcatcagagc gcttttggtt ttcaaaagcg ctctgaagtt
     6601 cctatacttt ctagctagag aataggaact tcccgagcgg ccgcgtgtta caagcttgac
     6661 tccagcgtaa ctggactgca atcaactcac tggctcacct tcacgggtgg gcctttcttc
     6721 ggtagaaaat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     6781 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     6841 tttttccgag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     6901 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     6961 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     7021 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     7081 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     7141 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     7201 aacaggagag cgcacgaggg agcttccagg ggtaaacgcc tggtatcttt atagtcctgt
     7261 cgggtttcgc cacctctgac ttgagcatcg atttttgtga tgctcgtcag gggggcggag
     7321 cctatggaaa aacgccagca acgcagaaag gcccacccga aggtgagcca ggtgattaca
     7381 tttgggccct catcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta
     7441 aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat ccgcgtccag
     7501 ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag
     7561 ggcggttttt tcctgtttgg tcacttacca atgcttaatc agtgaggcac ctatctcagc
     7621 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     7681 gcgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     7741 ggctccagat ttatcagcaa taaaccagcc agccgggagg gccgagcgca gaagtgatcc
     7801 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     7861 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     7921 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     7981 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgccagaag
     8041 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     8101 catgccatcc gtgagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     8161 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     8221 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gtaaactctc
     8281 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     8341 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     8401 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcattttag cttccttagc
     8461 tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg
     8521 aaagttggaa cctcttacgt gccgatcaag tcaaaagcct ccggtcggag gcttttgact
     8581 ttctgctatg gaggtcaggt atgatttaaa tggtcagtta ga
//
