LOCUS       sequence-291429-        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-m4046574
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 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 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
//
