LOCUS       sequence-297567-        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-m4046299
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 ttcgtctttt tagttttgct tccattggtg agctcccaat gcgttaatct gacaactcgc
      541 acgcagctac ctccagcata cactaactct ttcaccagag gcgtctatta ccccgacaaa
      601 gtatttcgtt cttcggtttt acatagtacc caagatttgt tcttgccatt cttttcaaac
      661 gttacttggt ttcacgccat acacgtttct ggtaccaatg gtactaagag gttcgataac
      721 ccagtgttgc cattcaatga cggtgtctac ttcgcttcca cagaaaagtc taacatcatt
      781 agaggttgga tctttggaac cacgcttgat tccaagactc aatctttgtt aattgtcaac
      841 aacgctacta atgttgtaat taaagtctgt gagttccaat tctgtaacga cccgtttttg
      901 ggtgtttatt accataagaa taataaaagc tggatggaat ccgaatttcg ggtttattca
      961 agtgctaaca actgtacctt cgaatacgtc agtcaacctt tcctgatgga tttggagggt
     1021 aagcaaggca acttcaagaa cttgagagaa tttgtcttca aaaatatcga cggttacttt
     1081 aagatctact ctaagcacac accaattaac ctagttagag atttacctca gggtttctct
     1141 gctttggaac cactcgttga tttgccgatt ggtatcaaca ttacgagatt tcaaactttg
     1201 ctagccttac accgatcata tttaactcct ggtgactcct cgtctggatg gactgcgggt
     1261 gctgctgcct actatgtcgg ttatttgcag ccaagaacct ttctgctcaa gtacaatgaa
     1321 aacgggacaa ttaccgatgc tgtagactgt gctttggatc cattgagcga aactaagtgc
     1381 accctaaaat ctttcacagt tgaaaagggt atataccaaa ctagcaactt tagagttcaa
     1441 ccaactgaat caatagtcag attcccaaat atcaccaact tgtgtccatt cggtgaggtt
     1501 tttaatgcca caaggtttgc ttctgtgtac gcttggaacc gtaagagaat ctccaattgt
     1561 gtggcagatt actctgtact atataacagt gcttccttct ctactttcaa atgttacggg
     1621 gtttcgccaa ccaagcttaa cgacttatgc ttcactaatg tttacgcaga ctcctttgtc
     1681 attagaggcg atgaagtaag acaaatcgcg ccaggtcaaa ccggtaagat tgctgattac
     1741 aactacaaat tgccagacga cttcaccggt tgcgtgatcg cttggaatag caacaacttg
     1801 gattcgaagg ttggtgggaa ctataactat ttgtaccgcc tatttcgtaa gtctaatttg
     1861 aaacctttcg aaagagacat aagtaccgag atttatcaag ctggttctac tccttgtaat
     1921 ggggtcgaag gttttaactg ttacttcccc ttacaatctt acggttttca gccaaccaac
     1981 ggagttggtt accaaccata tagagttgtt gtcctgtctt ttgaactatt gcatgcccca
     2041 gctacagtct gcggtcccaa aaagtccacg aacttggtta aaaataagtg tgtcaacttc
     2101 aatttcaatg gtcttactgg tactggcgta ttgaccgaat ccaacaagaa gttcctgcca
     2161 ttccaacaat tcggtagaga tattgcagat acgaccgatg cggtccggga tcctcaaact
     2221 ttggaaattt tggacatcac tccatgttcc tttggtggtg ttagcgttat tacccctgga
     2281 accaacactt ctaatcaggt cgccgtttta taccaagacg tcaactgtac ggaagtgcca
     2341 gttgctatcc acgctgacca actaacccct acttggagag tctactctac tggttctaat
     2401 gttttccaaa caagagctgg ttgtttgata ggtgctgagc atgtcaacaa ctcctatgaa
     2461 tgtgacattc caattggtgc cggaatctgt gcatcctacc aaacacagac taattcacca
     2521 agaagagcta gaagtgttgc ttcacaaagt atcattgcat acaccatgag cctaggtgcc
     2581 gaaaactcag tcgcatacag taacaattct attgctatac caactaattt cactatttct
     2641 gttactactg aaatcttacc tgtctctatg acaaagacct cagtggattg tacaatgtat
     2701 atctgtggtg actctactga gtgttctaat ttgttgttac aatacggttc cttctgcaca
     2761 caattgaacc gtgctcttac gggtattgct gtcgaacaag acaaaaacac ccaggaagta
     2821 tttgcccaag tcaagcaaat ctacaagacc ccacccatta aggatttcgg cggtttcaac
     2881 tttagccaaa tcttgccaga tccatcaaag ccatccaaga gatcgtttat cgaagattta
     2941 ttatttaata aagttacgtt agccgacgct ggattcatca agcaatatgg cgattgcttg
     3001 ggtgacattg cagcccgaga cttgatatgt gcgcagaagt tcaatggtct taccgtgttg
     3061 cccccactat tgactgatga aatgatcgct caatatacct cagcgctctt ggctgggacg
     3121 attacttctg gttggacatt tggtgctggt gctgcattgc aaatcccatt tgctatgcaa
     3181 atggcttaca gattcaacgg gataggagtt actcaaaacg ttctgtatga aaaccaaaag
     3241 ctaatcgcta atcaattcaa ctcggccatt ggtaagattc aagactcttt gtcctcgact
     3301 gcttcggcgc ttggtaaact acaagacgtt gttaaccaga atgctcaagc tttgaacacc
     3361 ctcgtaaaac agttgtcctc taacttcggg gccatttctt ccgtactcaa cgatatctta
     3421 tctaggttgg ataaggttga ggccgaagta caaatcgata gattgatcac cggaagattg
     3481 caatcgctac aaacctacgt tactcaacaa cttatccgcg ctgctgaaat tagagctagt
     3541 gctaatttag ccgcgaccaa gatgagtgaa tgtgtcctag gtcagtctaa gcgtgttgac
     3601 ttctgtggta aaggttatca tttgatgtcc tttccccaat ctgccccgca cggtgtcgtc
     3661 tttttgcatg ttacttacgt gccggctcag gaaaagaact tcactacagc tcctgccatc
     3721 tgtcacgatg gtaaagctca cttcccaaga gaaggtgttt tcgtttctaa tggtactcac
     3781 tggttcgtca cacaaagaaa cttctacgag ccccaaatta ttactactga caacaccttc
     3841 gtatccggta attgtgacgt tgtcatcggt attgtcaaca acaccgttta tgatccactc
     3901 caaccagaat tagattcctt caaggaagaa ttggataaat actttaagaa tcacacctcc
     3961 cctgacgttg atctaggtga catatctggc attaacgctt ccgtggttaa cattcaaaag
     4021 gaaatagaca ggttaaatga ggttgccaag aacttgaacg agtcattgat tgatctccaa
     4081 gaactaggta agtacgaaca atacattaaa tggccgtggt acatctggtt gggttttatc
     4141 gcagggctga ttgctattgt tatggtgacc atcatgttgt gctgtatgac ctcatgttgt
     4201 tcttgtttaa agggttgttg cagctgcggt tcttgttgca agttcgatga agacgactct
     4261 gaacctgtat tgaagggtgt taaattgcac tacacgctcg 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
//
