LOCUS       sequence-297202-        8670 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 surface glycoprotein (Spike 
            protein).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8670)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 8670)
  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-m4046464
FEATURES             Location/Qualifiers
     source          1..8670
                     /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..4380
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(4425..4443)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             4794..5597
                     /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      5680..6662
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      6740..6769
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      6791..7378
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(7390..7417)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(7441..7632)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(7633..8424,8425..8493))
                     /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     8256..8275
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8494..8596)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(8597..8628)
                     /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 ttctgattat
     4321 aaagatcatg atggtgatta taaagatcat gatattgatt ataaagatga tgatgataaa
     4381 taaacaggcc ccttttcctt tgtcgatatc atgtaattag ttatgtcacg cttacattca
     4441 cgccctcctc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc tgaagtctag
     4501 gtccctattt atttttttta atagttatgt tagtattaag aacgttattt atatttcaaa
     4561 tttttctttt ttttctgtac aaagcggccg ccgaaccact agtgcactgc agtacacggt
     4621 ttccttgaaa tttttttgat tcggtaatct ccgaacagaa ggaagaacga aggaaggagc
     4681 acagacttag attggtatat atacgcatat gtagtgttga agaaacatga aattgcccag
     4741 tattcttaac ccaactgcac agaacaaaaa cgtgcaggaa acgaagataa atcatgtcga
     4801 aagctacata taaggaacgt gctgctactc atcctagtcc tgttgctgcc aagctattta
     4861 atatcatgca cgaaaagcaa acaaacttgt gtgcttcatt ggatgttcgt accaccaagg
     4921 aattactgga gttagttgaa gcattaggtc ccaaaatttg tttactaaaa acacatgtgg
     4981 atatcttgac tgatttttcc atggagggca cagttaagcc gctaaaggca ttatccgcca
     5041 agtacaattt tttactcttc gaggacagaa aatttgctga cattggtaat acagtcaaat
     5101 tgcagtactc tgcgggtgta tacagaatag cagaatgggc agacattacg aatgcacacg
     5161 gtgtggtggg cccaggtatt gttagcggtt tgaagcaggc ggcagaagaa gtaacaaagg
     5221 aacctagagg ccttttgatg ttagcagaat tgtcatgcaa gggctcccta tctactggag
     5281 aatatactaa gggtactgtt gacattgcga agagcgacaa agattttgtt atcggcttta
     5341 ttgctcaaag agacatgggt ggaagagatg aaggttacga ttggttgatt atgacacccg
     5401 gtgtgggttt agatgacaag ggagatgcat tgggtcaaca gtatagaacc gtggatgatg
     5461 tggtttctac aggatctgac attattattg ttggaagagg actatttgca aagggaaggg
     5521 atgctaaggt agagggtgaa cgttacagaa aagcaggctg ggaagcatat ttgagaagat
     5581 gcggccagca aaactaaaaa actgtattat aagtaaatgc atgtatacta aactcacaaa
     5641 ttagagcttc aatttaatta tatcagttat tacccgagta acgaagcatc tgtgcttcat
     5701 tttgtagaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc
     5761 atttttacag aacagaaatg caacgcgaaa gcgctatttt accaacgaag aatctgtgct
     5821 tcatttttgt aaaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca aagaatctga
     5881 gctgcatttt tacagaacag aaatgcaacg cgagagcgct attttaccaa caaagaatct
     5941 atacttcttt tttgttctac aaaaatgcat cccgagagcg ctatttttct aacaaagcat
     6001 cttagattac tttttttctc ctttgtgcgc tctataatgc agtctcttga taactttttg
     6061 cactgtaggt ccgttaaggt tagaagaagg ctactttggt gtctattttc tcttccataa
     6121 aaaaagcctg actccacttc ccgcgtttac tgattactag cgaagctgcg ggtgcatttt
     6181 ttcaagataa aggcatcccc gattatattc tataccgatg tggattgcgc atactttgtg
     6241 aacagaaagt gatagcgttg atgattcttc attggtcaga aaattatgaa cggtttcttc
     6301 tattttgtct ctatatacta cgtataggaa atgtttacat tttcgtattg ttttcgattc
     6361 actctatgaa tagttcttac tacaattttt ttgtctaaag agtaatacta gagataaaca
     6421 taaaaaatgt agaggtcgag tttagatgca agttcaagga gcgaaaggtg gatgggtagg
     6481 ttatataggg atatagcaca gagatatata gcaaagagat acttttgagc aatgtttgtg
     6541 gaagcggtat tcgcaatatt ttagtagctc gttacagtcc ggtgcgtttt tggttttttg
     6601 aaagtgcgtc atcagagcgc ttttggtttt caaaagcgct ctgaagttcc tatactttct
     6661 agctagagaa taggaacttc ccgagcggcc gcgtgttaca agcttgactc cagcgtaact
     6721 ggactgcaat caactcactg gctcaccttc acgggtgggc ctttcttcgg tagaaaatca
     6781 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     6841 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaggt
     6901 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     6961 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     7021 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     7081 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     7141 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     7201 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     7261 cacgagggag cttccagggg taaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     7321 cctctgactt gagcatcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     7381 cgccagcaac gcagaaaggc ccacccgaag gtgagccagg tgattacatt tgggccctca
     7441 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc
     7501 gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt
     7561 ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc
     7621 ctgtttggtc acttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     7681 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatgc gggagggctt
     7741 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     7801 atcagcaata aaccagccag ccgggagggc cgagcgcaga agtgatcctg caactttatc
     7861 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     7921 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     7981 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     8041 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gccagaagta agttggccgc
     8101 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     8161 gagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     8221 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     8281 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcgt aaactctcaa ggatcttacc
     8341 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     8401 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     8461 aataagggcg acacggaaat gttgaatact cattttagct tccttagctc ctgaaaatct
     8521 cgataactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa agttggaacc
     8581 tcttacgtgc cgatcaagtc aaaagcctcc ggtcggaggc ttttgacttt ctgctatgga
     8641 ggtcaggtat gatttaaatg gtcagttaga
//
