LOCUS       sequence-297628-        8628 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 8628)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 8628)
  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-m4046409
FEATURES             Location/Qualifiers
     source          1..8628
                     /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..4338
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(4383..4401)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             4752..5555
                     /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      5638..6620
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      6698..6727
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      6749..7336
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(7348..7375)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(7399..7590)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(7591..8382,8383..8451))
                     /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     8214..8233
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8452..8554)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(8555..8586)
                     /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 ttcttggagc
     4321 catccgcagt ttgaaaaata aacaggcccc ttttcctttg tcgatatcat gtaattagtt
     4381 atgtcacgct tacattcacg ccctcctccc acatccgctc taaccgaaaa ggaaggagtt
     4441 agacaacctg aagtctaggt ccctatttat tttttttaat agttatgtta gtattaagaa
     4501 cgttatttat atttcaaatt tttctttttt ttctgtacaa agcggccgcc gaaccactag
     4561 tgcactgcag tacacggttt ccttgaaatt tttttgattc ggtaatctcc gaacagaagg
     4621 aagaacgaag gaaggagcac agacttagat tggtatatat acgcatatgt agtgttgaag
     4681 aaacatgaaa ttgcccagta ttcttaaccc aactgcacag aacaaaaacg tgcaggaaac
     4741 gaagataaat catgtcgaaa gctacatata aggaacgtgc tgctactcat cctagtcctg
     4801 ttgctgccaa gctatttaat atcatgcacg aaaagcaaac aaacttgtgt gcttcattgg
     4861 atgttcgtac caccaaggaa ttactggagt tagttgaagc attaggtccc aaaatttgtt
     4921 tactaaaaac acatgtggat atcttgactg atttttccat ggagggcaca gttaagccgc
     4981 taaaggcatt atccgccaag tacaattttt tactcttcga ggacagaaaa tttgctgaca
     5041 ttggtaatac agtcaaattg cagtactctg cgggtgtata cagaatagca gaatgggcag
     5101 acattacgaa tgcacacggt gtggtgggcc caggtattgt tagcggtttg aagcaggcgg
     5161 cagaagaagt aacaaaggaa cctagaggcc ttttgatgtt agcagaattg tcatgcaagg
     5221 gctccctatc tactggagaa tatactaagg gtactgttga cattgcgaag agcgacaaag
     5281 attttgttat cggctttatt gctcaaagag acatgggtgg aagagatgaa ggttacgatt
     5341 ggttgattat gacacccggt gtgggtttag atgacaaggg agatgcattg ggtcaacagt
     5401 atagaaccgt ggatgatgtg gtttctacag gatctgacat tattattgtt ggaagaggac
     5461 tatttgcaaa gggaagggat gctaaggtag agggtgaacg ttacagaaaa gcaggctggg
     5521 aagcatattt gagaagatgc ggccagcaaa actaaaaaac tgtattataa gtaaatgcat
     5581 gtatactaaa ctcacaaatt agagcttcaa tttaattata tcagttatta cccgagtaac
     5641 gaagcatctg tgcttcattt tgtagaacaa aaatgcaacg cgagagcgct aatttttcaa
     5701 acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgaaagc gctattttac
     5761 caacgaagaa tctgtgcttc atttttgtaa aacaaaaatg caacgcgaga gcgctaattt
     5821 ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg agagcgctat
     5881 tttaccaaca aagaatctat acttcttttt tgttctacaa aaatgcatcc cgagagcgct
     5941 atttttctaa caaagcatct tagattactt tttttctcct ttgtgcgctc tataatgcag
     6001 tctcttgata actttttgca ctgtaggtcc gttaaggtta gaagaaggct actttggtgt
     6061 ctattttctc ttccataaaa aaagcctgac tccacttccc gcgtttactg attactagcg
     6121 aagctgcggg tgcatttttt caagataaag gcatccccga ttatattcta taccgatgtg
     6181 gattgcgcat actttgtgaa cagaaagtga tagcgttgat gattcttcat tggtcagaaa
     6241 attatgaacg gtttcttcta ttttgtctct atatactacg tataggaaat gtttacattt
     6301 tcgtattgtt ttcgattcac tctatgaata gttcttacta caattttttt gtctaaagag
     6361 taatactaga gataaacata aaaaatgtag aggtcgagtt tagatgcaag ttcaaggagc
     6421 gaaaggtgga tgggtaggtt atatagggat atagcacaga gatatatagc aaagagatac
     6481 ttttgagcaa tgtttgtgga agcggtattc gcaatatttt agtagctcgt tacagtccgg
     6541 tgcgtttttg gttttttgaa agtgcgtcat cagagcgctt ttggttttca aaagcgctct
     6601 gaagttccta tactttctag ctagagaata ggaacttccc gagcggccgc gtgttacaag
     6661 cttgactcca gcgtaactgg actgcaatca actcactggc tcaccttcac gggtgggcct
     6721 ttcttcggta gaaaatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     6781 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     6841 caactctttt tccgaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     6901 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     6961 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     7021 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     7081 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     7141 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     7201 ggtcggaaca ggagagcgca cgagggagct tccaggggta aacgcctggt atctttatag
     7261 tcctgtcggg tttcgccacc tctgacttga gcatcgattt ttgtgatgct cgtcaggggg
     7321 gcggagccta tggaaaaacg ccagcaacgc agaaaggccc acccgaaggt gagccaggtg
     7381 attacatttg ggccctcatc agaggttttc accgtcatca ccgaaacgcg cgaggcagct
     7441 gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc
     7501 gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat
     7561 gttaagggcg gttttttcct gtttggtcac ttaccaatgc ttaatcagtg aggcacctat
     7621 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     7681 tacgatgcgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     7741 ctcaccggct ccagatttat cagcaataaa ccagccagcc gggagggccg agcgcagaag
     7801 tgatcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     7861 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     7921 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7981 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgc
     8041 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     8101 tactgtcatg ccatccgtga gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     8161 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     8221 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgtaa
     8281 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     8341 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     8401 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca ttttagcttc
     8461 cttagctcct gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc ttatttcatt
     8521 atggtgaaag ttggaacctc ttacgtgccg atcaagtcaa aagcctccgg tcggaggctt
     8581 ttgactttct gctatggagg tcaggtatga tttaaatggt cagttaga
//
