LOCUS       sequence-291550-        8581 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 8581)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 8581)
  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-m4046565
FEATURES             Location/Qualifiers
     source          1..8581
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             4250..4267
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             4268..4291
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(4336..4354)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             4705..5508
                     /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      5591..6573
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      6651..6680
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      6702..7289
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(7301..7328)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(7352..7543)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(7544..8335,8336..8404))
                     /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     8167..8186
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8405..8507)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(8508..8539)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgccc cacacaccat agcttcaaaa tgtttctact ccttttttac tcttccagat
       61 tttctcggac tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag catactaaat
      121 ttcccctctt tcttcctcta gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa
      181 aaaagagacc gcctcgtttc tttttcttcg tcgaaaaagg caataaaaat ttttatcacg
      241 tttctttttc ttgaaaattt ttttttttga tttttttctc tttcgatgac ctcccattga
      301 tatttaagtt aataaacggt cttcaatttc tcaagtttca gtttcatttt tcttgttcta
      361 ttacaacttt ttttacttct tgctcattag aaagaaagca tagcaatcta atctaagttt
      421 taattacaaa atgttcgtct ttttagtttt gcttccattg gtgagctccc aatgcgttaa
      481 tctgacaact cgcacgcagc tacctccagc atacactaac tctttcacca gaggcgtcta
      541 ttaccccgac aaagtatttc gttcttcggt tttacatagt acccaagatt tgttcttgcc
      601 attcttttca aacgttactt ggtttcacgc catacacgtt tctggtacca atggtactaa
      661 gaggttcgat aacccagtgt tgccattcaa tgacggtgtc tacttcgctt ccacagaaaa
      721 gtctaacatc attagaggtt ggatctttgg aaccacgctt gattccaaga ctcaatcttt
      781 gttaattgtc aacaacgcta ctaatgttgt aattaaagtc tgtgagttcc aattctgtaa
      841 cgacccgttt ttgggtgttt attaccataa gaataataaa agctggatgg aatccgaatt
      901 tcgggtttat tcaagtgcta acaactgtac cttcgaatac gtcagtcaac ctttcctgat
      961 ggatttggag ggtaagcaag gcaacttcaa gaacttgaga gaatttgtct tcaaaaatat
     1021 cgacggttac tttaagatct actctaagca cacaccaatt aacctagtta gagatttacc
     1081 tcagggtttc tctgctttgg aaccactcgt tgatttgccg attggtatca acattacgag
     1141 atttcaaact ttgctagcct tacaccgatc atatttaact cctggtgact cctcgtctgg
     1201 atggactgcg ggtgctgctg cctactatgt cggttatttg cagccaagaa cctttctgct
     1261 caagtacaat gaaaacggga caattaccga tgctgtagac tgtgctttgg atccattgag
     1321 cgaaactaag tgcaccctaa aatctttcac agttgaaaag ggtatatacc aaactagcaa
     1381 ctttagagtt caaccaactg aatcaatagt cagattccca aatatcacca acttgtgtcc
     1441 attcggtgag gtttttaatg ccacaaggtt tgcttctgtg tacgcttgga accgtaagag
     1501 aatctccaat tgtgtggcag attactctgt actatataac agtgcttcct tctctacttt
     1561 caaatgttac ggggtttcgc caaccaagct taacgactta tgcttcacta atgtttacgc
     1621 agactccttt gtcattagag gcgatgaagt aagacaaatc gcgccaggtc aaaccggtaa
     1681 gattgctgat tacaactaca aattgccaga cgacttcacc ggttgcgtga tcgcttggaa
     1741 tagcaacaac ttggattcga aggttggtgg gaactataac tatttgtacc gcctatttcg
     1801 taagtctaat ttgaaacctt tcgaaagaga cataagtacc gagatttatc aagctggttc
     1861 tactccttgt aatggggtcg aaggttttaa ctgttacttc cccttacaat cttacggttt
     1921 tcagccaacc aacggagttg gttaccaacc atatagagtt gttgtcctgt cttttgaact
     1981 attgcatgcc ccagctacag tctgcggtcc caaaaagtcc acgaacttgg ttaaaaataa
     2041 gtgtgtcaac ttcaatttca atggtcttac tggtactggc gtattgaccg aatccaacaa
     2101 gaagttcctg ccattccaac aattcggtag agatattgca gatacgaccg atgcggtccg
     2161 ggatcctcaa actttggaaa ttttggacat cactccatgt tcctttggtg gtgttagcgt
     2221 tattacccct ggaaccaaca cttctaatca ggtcgccgtt ttataccaag acgtcaactg
     2281 tacggaagtg ccagttgcta tccacgctga ccaactaacc cctacttgga gagtctactc
     2341 tactggttct aatgttttcc aaacaagagc tggttgtttg ataggtgctg agcatgtcaa
     2401 caactcctat gaatgtgaca ttccaattgg tgccggaatc tgtgcatcct accaaacaca
     2461 gactaattca ccaagaagag ctagaagtgt tgcttcacaa agtatcattg catacaccat
     2521 gagcctaggt gccgaaaact cagtcgcata cagtaacaat tctattgcta taccaactaa
     2581 tttcactatt tctgttacta ctgaaatctt acctgtctct atgacaaaga cctcagtgga
     2641 ttgtacaatg tatatctgtg gtgactctac tgagtgttct aatttgttgt tacaatacgg
     2701 ttccttctgc acacaattga accgtgctct tacgggtatt gctgtcgaac aagacaaaaa
     2761 cacccaggaa gtatttgccc aagtcaagca aatctacaag accccaccca ttaaggattt
     2821 cggcggtttc aactttagcc aaatcttgcc agatccatca aagccatcca agagatcgtt
     2881 tatcgaagat ttattattta ataaagttac gttagccgac gctggattca tcaagcaata
     2941 tggcgattgc ttgggtgaca ttgcagcccg agacttgata tgtgcgcaga agttcaatgg
     3001 tcttaccgtg ttgcccccac tattgactga tgaaatgatc gctcaatata cctcagcgct
     3061 cttggctggg acgattactt ctggttggac atttggtgct ggtgctgcat tgcaaatccc
     3121 atttgctatg caaatggctt acagattcaa cgggatagga gttactcaaa acgttctgta
     3181 tgaaaaccaa aagctaatcg ctaatcaatt caactcggcc attggtaaga ttcaagactc
     3241 tttgtcctcg actgcttcgg cgcttggtaa actacaagac gttgttaacc agaatgctca
     3301 agctttgaac accctcgtaa aacagttgtc ctctaacttc ggggccattt cttccgtact
     3361 caacgatatc ttatctaggt tggataaggt tgaggccgaa gtacaaatcg atagattgat
     3421 caccggaaga ttgcaatcgc tacaaaccta cgttactcaa caacttatcc gcgctgctga
     3481 aattagagct agtgctaatt tagccgcgac caagatgagt gaatgtgtcc taggtcagtc
     3541 taagcgtgtt gacttctgtg gtaaaggtta tcatttgatg tcctttcccc aatctgcccc
     3601 gcacggtgtc gtctttttgc atgttactta cgtgccggct caggaaaaga acttcactac
     3661 agctcctgcc atctgtcacg atggtaaagc tcacttccca agagaaggtg ttttcgtttc
     3721 taatggtact cactggttcg tcacacaaag aaacttctac gagccccaaa ttattactac
     3781 tgacaacacc ttcgtatccg gtaattgtga cgttgtcatc ggtattgtca acaacaccgt
     3841 ttatgatcca ctccaaccag aattagattc cttcaaggaa gaattggata aatactttaa
     3901 gaatcacacc tcccctgacg ttgatctagg tgacatatct ggcattaacg cttccgtggt
     3961 taacattcaa aaggaaatag acaggttaaa tgaggttgcc aagaacttga acgagtcatt
     4021 gattgatctc caagaactag gtaagtacga acaatacatt aaatggccgt ggtacatctg
     4081 gttgggtttt atcgcagggc tgattgctat tgttatggtg accatcatgt tgtgctgtat
     4141 gacctcatgt tgttcttgtt taaagggttg ttgcagctgc ggttcttgtt gcaagttcga
     4201 tgaagacgac tctgaacctg tattgaaggg tgttaaattg cactacacgc tcgtgccgag
     4261 aggttcttgg agccatccgc agtttgaaaa ataaacaggc cccttttcct ttgtcgatat
     4321 catgtaatta gttatgtcac gcttacattc acgccctcct cccacatccg ctctaaccga
     4381 aaaggaagga gttagacaac ctgaagtcta ggtccctatt tatttttttt aatagttatg
     4441 ttagtattaa gaacgttatt tatatttcaa atttttcttt tttttctgta caaagcggcc
     4501 gccgaaccac tagtgcactg cagtacacgg tttccttgaa atttttttga ttcggtaatc
     4561 tccgaacaga aggaagaacg aaggaaggag cacagactta gattggtata tatacgcata
     4621 tgtagtgttg aagaaacatg aaattgccca gtattcttaa cccaactgca cagaacaaaa
     4681 acgtgcagga aacgaagata aatcatgtcg aaagctacat ataaggaacg tgctgctact
     4741 catcctagtc ctgttgctgc caagctattt aatatcatgc acgaaaagca aacaaacttg
     4801 tgtgcttcat tggatgttcg taccaccaag gaattactgg agttagttga agcattaggt
     4861 cccaaaattt gtttactaaa aacacatgtg gatatcttga ctgatttttc catggagggc
     4921 acagttaagc cgctaaaggc attatccgcc aagtacaatt ttttactctt cgaggacaga
     4981 aaatttgctg acattggtaa tacagtcaaa ttgcagtact ctgcgggtgt atacagaata
     5041 gcagaatggg cagacattac gaatgcacac ggtgtggtgg gcccaggtat tgttagcggt
     5101 ttgaagcagg cggcagaaga agtaacaaag gaacctagag gccttttgat gttagcagaa
     5161 ttgtcatgca agggctccct atctactgga gaatatacta agggtactgt tgacattgcg
     5221 aagagcgaca aagattttgt tatcggcttt attgctcaaa gagacatggg tggaagagat
     5281 gaaggttacg attggttgat tatgacaccc ggtgtgggtt tagatgacaa gggagatgca
     5341 ttgggtcaac agtatagaac cgtggatgat gtggtttcta caggatctga cattattatt
     5401 gttggaagag gactatttgc aaagggaagg gatgctaagg tagagggtga acgttacaga
     5461 aaagcaggct gggaagcata tttgagaaga tgcggccagc aaaactaaaa aactgtatta
     5521 taagtaaatg catgtatact aaactcacaa attagagctt caatttaatt atatcagtta
     5581 ttacccgagt aacgaagcat ctgtgcttca ttttgtagaa caaaaatgca acgcgagagc
     5641 gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgaa
     5701 agcgctattt taccaacgaa gaatctgtgc ttcatttttg taaaacaaaa atgcaacgcg
     5761 agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac
     5821 gcgagagcgc tattttacca acaaagaatc tatacttctt ttttgttcta caaaaatgca
     5881 tcccgagagc gctatttttc taacaaagca tcttagatta ctttttttct cctttgtgcg
     5941 ctctataatg cagtctcttg ataacttttt gcactgtagg tccgttaagg ttagaagaag
     6001 gctactttgg tgtctatttt ctcttccata aaaaaagcct gactccactt cccgcgttta
     6061 ctgattacta gcgaagctgc gggtgcattt tttcaagata aaggcatccc cgattatatt
     6121 ctataccgat gtggattgcg catactttgt gaacagaaag tgatagcgtt gatgattctt
     6181 cattggtcag aaaattatga acggtttctt ctattttgtc tctatatact acgtatagga
     6241 aatgtttaca ttttcgtatt gttttcgatt cactctatga atagttctta ctacaatttt
     6301 tttgtctaaa gagtaatact agagataaac ataaaaaatg tagaggtcga gtttagatgc
     6361 aagttcaagg agcgaaaggt ggatgggtag gttatatagg gatatagcac agagatatat
     6421 agcaaagaga tacttttgag caatgtttgt ggaagcggta ttcgcaatat tttagtagct
     6481 cgttacagtc cggtgcgttt ttggtttttt gaaagtgcgt catcagagcg cttttggttt
     6541 tcaaaagcgc tctgaagttc ctatactttc tagctagaga ataggaactt cccgagcggc
     6601 cgcgtgttac aagcttgact ccagcgtaac tggactgcaa tcaactcact ggctcacctt
     6661 cacgggtggg cctttcttcg gtagaaaatc aaaggatctt cttgagatcc tttttttctg
     6721 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     6781 gatcaagagc taccaactct ttttccgagg taactggctt cagcagagcg cagataccaa
     6841 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     6901 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     6961 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     7021 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     7081 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     7141 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg gtaaacgcct
     7201 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcatcga tttttgtgat
     7261 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcagaaagg cccacccgaa
     7321 ggtgagccag gtgattacat ttgggccctc atcagaggtt ttcaccgtca tcaccgaaac
     7381 gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg
     7441 cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga
     7501 taaagcgggc catgttaagg gcggtttttt cctgtttggt cacttaccaa tgcttaatca
     7561 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     7621 tcgtgtagat aactacgatg cgggagggct taccatctgg ccccagtgct gcaatgatac
     7681 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccgggaggg
     7741 ccgagcgcag aagtgatcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     7801 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     7861 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7921 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7981 ctccgatcgt tgccagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     8041 tgcataattc tcttactgtc atgccatccg tgagatgctt ttctgtgact ggtgagtact
     8101 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     8161 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     8221 cttcggggcg taaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     8281 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     8341 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     8401 tcattttagc ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg
     8461 atcttatttc attatggtga aagttggaac ctcttacgtg ccgatcaagt caaaagcctc
     8521 cggtcggagg cttttgactt tctgctatgg aggtcaggta tgatttaaat ggtcagttag
     8581 a
//
