LOCUS       sequence-297579-        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-m4046350
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 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 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
//
