LOCUS       sequence-300432-        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"
     terminator      997..1026
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      1048..1635
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(1647..1674)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1698..1889)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(1890..2681,2682..2750))
                     /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     2513..2532
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2751..2853)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(2854..2885)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        2952..3393
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    2952..3069
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     3364..3387
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             7224..7241
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             7242..7265
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(7310..7328)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             7679..8482
                     /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      join(8565..8628,1..919)
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgaaagcg ctattttacc
       61 aacgaagaat ctgtgcttca tttttgtaaa acaaaaatgc aacgcgagag cgctaatttt
      121 tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga gagcgctatt
      181 ttaccaacaa agaatctata cttctttttt gttctacaaa aatgcatccc gagagcgcta
      241 tttttctaac aaagcatctt agattacttt ttttctcctt tgtgcgctct ataatgcagt
      301 ctcttgataa ctttttgcac tgtaggtccg ttaaggttag aagaaggcta ctttggtgtc
      361 tattttctct tccataaaaa aagcctgact ccacttcccg cgtttactga ttactagcga
      421 agctgcgggt gcattttttc aagataaagg catccccgat tatattctat accgatgtgg
      481 attgcgcata ctttgtgaac agaaagtgat agcgttgatg attcttcatt ggtcagaaaa
      541 ttatgaacgg tttcttctat tttgtctcta tatactacgt ataggaaatg tttacatttt
      601 cgtattgttt tcgattcact ctatgaatag ttcttactac aatttttttg tctaaagagt
      661 aatactagag ataaacataa aaaatgtaga ggtcgagttt agatgcaagt tcaaggagcg
      721 aaaggtggat gggtaggtta tatagggata tagcacagag atatatagca aagagatact
      781 tttgagcaat gtttgtggaa gcggtattcg caatatttta gtagctcgtt acagtccggt
      841 gcgtttttgg ttttttgaaa gtgcgtcatc agagcgcttt tggttttcaa aagcgctctg
      901 aagttcctat actttctagc tagagaatag gaacttcccg agcggccgcg tgttacaagc
      961 ttgactccag cgtaactgga ctgcaatcaa ctcactggct caccttcacg ggtgggcctt
     1021 tcttcggtag aaaatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     1081 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     1141 aactcttttt ccgaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     1201 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     1261 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     1321 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     1381 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     1441 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     1501 gtcggaacag gagagcgcac gagggagctt ccaggggtaa acgcctggta tctttatagt
     1561 cctgtcgggt ttcgccacct ctgacttgag catcgatttt tgtgatgctc gtcagggggg
     1621 cggagcctat ggaaaaacgc cagcaacgca gaaaggccca cccgaaggtg agccaggtga
     1681 ttacatttgg gccctcatca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg
     1741 cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg
     1801 tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg
     1861 ttaagggcgg ttttttcctg tttggtcact taccaatgct taatcagtga ggcacctatc
     1921 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     1981 acgatgcggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     2041 tcaccggctc cagatttatc agcaataaac cagccagccg ggagggccga gcgcagaagt
     2101 gatcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     2161 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     2221 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     2281 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgcc
     2341 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     2401 actgtcatgc catccgtgag atgcttttct gtgactggtg agtactcaac caagtcattc
     2461 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     2521 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgtaaa
     2581 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     2641 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     2701 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat tttagcttcc
     2761 ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct tatttcatta
     2821 tggtgaaagt tggaacctct tacgtgccga tcaagtcaaa agcctccggt cggaggcttt
     2881 tgactttctg ctatggaggt caggtatgat ttaaatggtc agttagagcg gccgcttgct
     2941 atattgaagt acggattaga agccgccgag cgggcgacag ccctccgacg gaagactctc
     3001 ctccgtgcgt cctcgtcttc accggtcgcg ttcctgaaac gcagatgtgc ctcgcgccgc
     3061 actgctccga acaataaaga ttctacaata ctagctttta tggttatgaa gaggaaaaat
     3121 tggcagtaac ctggccccac aaaccttcaa attaacgaat caaattaaca accataggat
     3181 gataatgcga ttagtttttt agccttattt ctggggtaat taatcagcga agcgatgatt
     3241 tttgatctat taacagatat ataaatggaa aagctgcata accactttaa ctaatacttt
     3301 caacattttc agtttgtatt acttcttatt caaatgtcat aaaagtatca acaaaaaatt
     3361 gttaatatac ctctatactt taacgtcaag gagaaaaaac tataatgttc gtctttttag
     3421 ttttgcttcc attggtgagc tcccaatgcg ttaatctgac aactcgcacg cagctacctc
     3481 cagcatacac taactctttc accagaggcg tctattaccc cgacaaagta tttcgttctt
     3541 cggttttaca tagtacccaa gatttgttct tgccattctt ttcaaacgtt acttggtttc
     3601 acgccataca cgtttctggt accaatggta ctaagaggtt cgataaccca gtgttgccat
     3661 tcaatgacgg tgtctacttc gcttccacag aaaagtctaa catcattaga ggttggatct
     3721 ttggaaccac gcttgattcc aagactcaat ctttgttaat tgtcaacaac gctactaatg
     3781 ttgtaattaa agtctgtgag ttccaattct gtaacgaccc gtttttgggt gtttattacc
     3841 ataagaataa taaaagctgg atggaatccg aatttcgggt ttattcaagt gctaacaact
     3901 gtaccttcga atacgtcagt caacctttcc tgatggattt ggagggtaag caaggcaact
     3961 tcaagaactt gagagaattt gtcttcaaaa atatcgacgg ttactttaag atctactcta
     4021 agcacacacc aattaaccta gttagagatt tacctcaggg tttctctgct ttggaaccac
     4081 tcgttgattt gccgattggt atcaacatta cgagatttca aactttgcta gccttacacc
     4141 gatcatattt aactcctggt gactcctcgt ctggatggac tgcgggtgct gctgcctact
     4201 atgtcggtta tttgcagcca agaacctttc tgctcaagta caatgaaaac gggacaatta
     4261 ccgatgctgt agactgtgct ttggatccat tgagcgaaac taagtgcacc ctaaaatctt
     4321 tcacagttga aaagggtata taccaaacta gcaactttag agttcaacca actgaatcaa
     4381 tagtcagatt cccaaatatc accaacttgt gtccattcgg tgaggttttt aatgccacaa
     4441 ggtttgcttc tgtgtacgct tggaaccgta agagaatctc caattgtgtg gcagattact
     4501 ctgtactata taacagtgct tccttctcta ctttcaaatg ttacggggtt tcgccaacca
     4561 agcttaacga cttatgcttc actaatgttt acgcagactc ctttgtcatt agaggcgatg
     4621 aagtaagaca aatcgcgcca ggtcaaaccg gtaagattgc tgattacaac tacaaattgc
     4681 cagacgactt caccggttgc gtgatcgctt ggaatagcaa caacttggat tcgaaggttg
     4741 gtgggaacta taactatttg taccgcctat ttcgtaagtc taatttgaaa cctttcgaaa
     4801 gagacataag taccgagatt tatcaagctg gttctactcc ttgtaatggg gtcgaaggtt
     4861 ttaactgtta cttcccctta caatcttacg gttttcagcc aaccaacgga gttggttacc
     4921 aaccatatag agttgttgtc ctgtcttttg aactattgca tgccccagct acagtctgcg
     4981 gtcccaaaaa gtccacgaac ttggttaaaa ataagtgtgt caacttcaat ttcaatggtc
     5041 ttactggtac tggcgtattg accgaatcca acaagaagtt cctgccattc caacaattcg
     5101 gtagagatat tgcagatacg accgatgcgg tccgggatcc tcaaactttg gaaattttgg
     5161 acatcactcc atgttccttt ggtggtgtta gcgttattac ccctggaacc aacacttcta
     5221 atcaggtcgc cgttttatac caagacgtca actgtacgga agtgccagtt gctatccacg
     5281 ctgaccaact aacccctact tggagagtct actctactgg ttctaatgtt ttccaaacaa
     5341 gagctggttg tttgataggt gctgagcatg tcaacaactc ctatgaatgt gacattccaa
     5401 ttggtgccgg aatctgtgca tcctaccaaa cacagactaa ttcaccaaga agagctagaa
     5461 gtgttgcttc acaaagtatc attgcataca ccatgagcct aggtgccgaa aactcagtcg
     5521 catacagtaa caattctatt gctataccaa ctaatttcac tatttctgtt actactgaaa
     5581 tcttacctgt ctctatgaca aagacctcag tggattgtac aatgtatatc tgtggtgact
     5641 ctactgagtg ttctaatttg ttgttacaat acggttcctt ctgcacacaa ttgaaccgtg
     5701 ctcttacggg tattgctgtc gaacaagaca aaaacaccca ggaagtattt gcccaagtca
     5761 agcaaatcta caagacccca cccattaagg atttcggcgg tttcaacttt agccaaatct
     5821 tgccagatcc atcaaagcca tccaagagat cgtttatcga agatttatta tttaataaag
     5881 ttacgttagc cgacgctgga ttcatcaagc aatatggcga ttgcttgggt gacattgcag
     5941 cccgagactt gatatgtgcg cagaagttca atggtcttac cgtgttgccc ccactattga
     6001 ctgatgaaat gatcgctcaa tatacctcag cgctcttggc tgggacgatt acttctggtt
     6061 ggacatttgg tgctggtgct gcattgcaaa tcccatttgc tatgcaaatg gcttacagat
     6121 tcaacgggat aggagttact caaaacgttc tgtatgaaaa ccaaaagcta atcgctaatc
     6181 aattcaactc ggccattggt aagattcaag actctttgtc ctcgactgct tcggcgcttg
     6241 gtaaactaca agacgttgtt aaccagaatg ctcaagcttt gaacaccctc gtaaaacagt
     6301 tgtcctctaa cttcggggcc atttcttccg tactcaacga tatcttatct aggttggata
     6361 aggttgaggc cgaagtacaa atcgatagat tgatcaccgg aagattgcaa tcgctacaaa
     6421 cctacgttac tcaacaactt atccgcgctg ctgaaattag agctagtgct aatttagccg
     6481 cgaccaagat gagtgaatgt gtcctaggtc agtctaagcg tgttgacttc tgtggtaaag
     6541 gttatcattt gatgtccttt ccccaatctg ccccgcacgg tgtcgtcttt ttgcatgtta
     6601 cttacgtgcc ggctcaggaa aagaacttca ctacagctcc tgccatctgt cacgatggta
     6661 aagctcactt cccaagagaa ggtgttttcg tttctaatgg tactcactgg ttcgtcacac
     6721 aaagaaactt ctacgagccc caaattatta ctactgacaa caccttcgta tccggtaatt
     6781 gtgacgttgt catcggtatt gtcaacaaca ccgtttatga tccactccaa ccagaattag
     6841 attccttcaa ggaagaattg gataaatact ttaagaatca cacctcccct gacgttgatc
     6901 taggtgacat atctggcatt aacgcttccg tggttaacat tcaaaaggaa atagacaggt
     6961 taaatgaggt tgccaagaac ttgaacgagt cattgattga tctccaagaa ctaggtaagt
     7021 acgaacaata cattaaatgg ccgtggtaca tctggttggg ttttatcgca gggctgattg
     7081 ctattgttat ggtgaccatc atgttgtgct gtatgacctc atgttgttct tgtttaaagg
     7141 gttgttgcag ctgcggttct tgttgcaagt tcgatgaaga cgactctgaa cctgtattga
     7201 agggtgttaa attgcactac acgctcgtgc cgagaggttc ttggagccat ccgcagtttg
     7261 aaaaataaac aggccccttt tcctttgtcg atatcatgta attagttatg tcacgcttac
     7321 attcacgccc tcctcccaca tccgctctaa ccgaaaagga aggagttaga caacctgaag
     7381 tctaggtccc tatttatttt ttttaatagt tatgttagta ttaagaacgt tatttatatt
     7441 tcaaattttt cttttttttc tgtacaaagc ggccgccgaa ccactagtgc actgcagtac
     7501 acggtttcct tgaaattttt ttgattcggt aatctccgaa cagaaggaag aacgaaggaa
     7561 ggagcacaga cttagattgg tatatatacg catatgtagt gttgaagaaa catgaaattg
     7621 cccagtattc ttaacccaac tgcacagaac aaaaacgtgc aggaaacgaa gataaatcat
     7681 gtcgaaagct acatataagg aacgtgctgc tactcatcct agtcctgttg ctgccaagct
     7741 atttaatatc atgcacgaaa agcaaacaaa cttgtgtgct tcattggatg ttcgtaccac
     7801 caaggaatta ctggagttag ttgaagcatt aggtcccaaa atttgtttac taaaaacaca
     7861 tgtggatatc ttgactgatt tttccatgga gggcacagtt aagccgctaa aggcattatc
     7921 cgccaagtac aattttttac tcttcgagga cagaaaattt gctgacattg gtaatacagt
     7981 caaattgcag tactctgcgg gtgtatacag aatagcagaa tgggcagaca ttacgaatgc
     8041 acacggtgtg gtgggcccag gtattgttag cggtttgaag caggcggcag aagaagtaac
     8101 aaaggaacct agaggccttt tgatgttagc agaattgtca tgcaagggct ccctatctac
     8161 tggagaatat actaagggta ctgttgacat tgcgaagagc gacaaagatt ttgttatcgg
     8221 ctttattgct caaagagaca tgggtggaag agatgaaggt tacgattggt tgattatgac
     8281 acccggtgtg ggtttagatg acaagggaga tgcattgggt caacagtata gaaccgtgga
     8341 tgatgtggtt tctacaggat ctgacattat tattgttgga agaggactat ttgcaaaggg
     8401 aagggatgct aaggtagagg gtgaacgtta cagaaaagca ggctgggaag catatttgag
     8461 aagatgcggc cagcaaaact aaaaaactgt attataagta aatgcatgta tactaaactc
     8521 acaaattaga gcttcaattt aattatatca gttattaccc gagtaacgaa gcatctgtgc
     8581 ttcattttgt agaacaaaaa tgcaacgcga gagcgctaat ttttcaaa
//
