LOCUS       sequence-307325-        8575 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 8575)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 8575)
  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-m4046494
FEATURES             Location/Qualifiers
     source          1..8575
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        18..425
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             4258..4275
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             4276..4293
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4338..4356)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             4707..5510
                     /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      5593..6575
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      6653..6682
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      6704..7291
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(7303..7330)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(7354..7545)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(7546..8337,8338..8406))
                     /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     8169..8188
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8407..8509)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(8510..8541)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 cagttagagc ggccgcccca cacaccatag cttcaaaatg tttctactcc ttttttactc
       61 ttccagattt tctcggactc cgcgcatcgc cgtaccactt caaaacaccc aagcacagca
      121 tactaaattt cccctctttc ttcctctagg gtgtcgttaa ttacccgtac taaaggtttg
      181 gaaaagaaaa aagagaccgc ctcgtttctt tttcttcgtc gaaaaaggca ataaaaattt
      241 ttatcacgtt tctttttctt gaaaattttt ttttttgatt tttttctctt tcgatgacct
      301 cccattgata tttaagttaa taaacggtct tcaatttctc aagtttcagt ttcatttttc
      361 ttgttctatt acaacttttt ttacttcttg ctcattagaa agaaagcata gcaatctaat
      421 ctaagtttta attacaaaat gttcgtcttt ttagttttgc ttccattggt gagctcccaa
      481 tgcgttaatc tgacaactcg cacgcagcta cctccagcat acactaactc tttcaccaga
      541 ggcgtctatt accccgacaa agtatttcgt tcttcggttt tacatagtac ccaagatttg
      601 ttcttgccat tcttttcaaa cgttacttgg tttcacgcca tacacgtttc tggtaccaat
      661 ggtactaaga ggttcgataa cccagtgttg ccattcaatg acggtgtcta cttcgcttcc
      721 acagaaaagt ctaacatcat tagaggttgg atctttggaa ccacgcttga ttccaagact
      781 caatctttgt taattgtcaa caacgctact aatgttgtaa ttaaagtctg tgagttccaa
      841 ttctgtaacg acccgttttt gggtgtttat taccataaga ataataaaag ctggatggaa
      901 tccgaatttc gggtttattc aagtgctaac aactgtacct tcgaatacgt cagtcaacct
      961 ttcctgatgg atttggaggg taagcaaggc aacttcaaga acttgagaga atttgtcttc
     1021 aaaaatatcg acggttactt taagatctac tctaagcaca caccaattaa cctagttaga
     1081 gatttacctc agggtttctc tgctttggaa ccactcgttg atttgccgat tggtatcaac
     1141 attacgagat ttcaaacttt gctagcctta caccgatcat atttaactcc tggtgactcc
     1201 tcgtctggat ggactgcggg tgctgctgcc tactatgtcg gttatttgca gccaagaacc
     1261 tttctgctca agtacaatga aaacgggaca attaccgatg ctgtagactg tgctttggat
     1321 ccattgagcg aaactaagtg caccctaaaa tctttcacag ttgaaaaggg tatataccaa
     1381 actagcaact ttagagttca accaactgaa tcaatagtca gattcccaaa tatcaccaac
     1441 ttgtgtccat tcggtgaggt ttttaatgcc acaaggtttg cttctgtgta cgcttggaac
     1501 cgtaagagaa tctccaattg tgtggcagat tactctgtac tatataacag tgcttccttc
     1561 tctactttca aatgttacgg ggtttcgcca accaagctta acgacttatg cttcactaat
     1621 gtttacgcag actcctttgt cattagaggc gatgaagtaa gacaaatcgc gccaggtcaa
     1681 accggtaaga ttgctgatta caactacaaa ttgccagacg acttcaccgg ttgcgtgatc
     1741 gcttggaata gcaacaactt ggattcgaag gttggtggga actataacta tttgtaccgc
     1801 ctatttcgta agtctaattt gaaacctttc gaaagagaca taagtaccga gatttatcaa
     1861 gctggttcta ctccttgtaa tggggtcgaa ggttttaact gttacttccc cttacaatct
     1921 tacggttttc agccaaccaa cggagttggt taccaaccat atagagttgt tgtcctgtct
     1981 tttgaactat tgcatgcccc agctacagtc tgcggtccca aaaagtccac gaacttggtt
     2041 aaaaataagt gtgtcaactt caatttcaat ggtcttactg gtactggcgt attgaccgaa
     2101 tccaacaaga agttcctgcc attccaacaa ttcggtagag atattgcaga tacgaccgat
     2161 gcggtccggg atcctcaaac tttggaaatt ttggacatca ctccatgttc ctttggtggt
     2221 gttagcgtta ttacccctgg aaccaacact tctaatcagg tcgccgtttt ataccaagac
     2281 gtcaactgta cggaagtgcc agttgctatc cacgctgacc aactaacccc tacttggaga
     2341 gtctactcta ctggttctaa tgttttccaa acaagagctg gttgtttgat aggtgctgag
     2401 catgtcaaca actcctatga atgtgacatt ccaattggtg ccggaatctg tgcatcctac
     2461 caaacacaga ctaattcacc aagaagagct agaagtgttg cttcacaaag tatcattgca
     2521 tacaccatga gcctaggtgc cgaaaactca gtcgcataca gtaacaattc tattgctata
     2581 ccaactaatt tcactatttc tgttactact gaaatcttac ctgtctctat gacaaagacc
     2641 tcagtggatt gtacaatgta tatctgtggt gactctactg agtgttctaa tttgttgtta
     2701 caatacggtt ccttctgcac acaattgaac cgtgctctta cgggtattgc tgtcgaacaa
     2761 gacaaaaaca cccaggaagt atttgcccaa gtcaagcaaa tctacaagac cccacccatt
     2821 aaggatttcg gcggtttcaa ctttagccaa atcttgccag atccatcaaa gccatccaag
     2881 agatcgttta tcgaagattt attatttaat aaagttacgt tagccgacgc tggattcatc
     2941 aagcaatatg gcgattgctt gggtgacatt gcagcccgag acttgatatg tgcgcagaag
     3001 ttcaatggtc ttaccgtgtt gcccccacta ttgactgatg aaatgatcgc tcaatatacc
     3061 tcagcgctct tggctgggac gattacttct ggttggacat ttggtgctgg tgctgcattg
     3121 caaatcccat ttgctatgca aatggcttac agattcaacg ggataggagt tactcaaaac
     3181 gttctgtatg aaaaccaaaa gctaatcgct aatcaattca actcggccat tggtaagatt
     3241 caagactctt tgtcctcgac tgcttcggcg cttggtaaac tacaagacgt tgttaaccag
     3301 aatgctcaag ctttgaacac cctcgtaaaa cagttgtcct ctaacttcgg ggccatttct
     3361 tccgtactca acgatatctt atctaggttg gataaggttg aggccgaagt acaaatcgat
     3421 agattgatca ccggaagatt gcaatcgcta caaacctacg ttactcaaca acttatccgc
     3481 gctgctgaaa ttagagctag tgctaattta gccgcgacca agatgagtga atgtgtccta
     3541 ggtcagtcta agcgtgttga cttctgtggt aaaggttatc atttgatgtc ctttccccaa
     3601 tctgccccgc acggtgtcgt ctttttgcat gttacttacg tgccggctca ggaaaagaac
     3661 ttcactacag ctcctgccat ctgtcacgat ggtaaagctc acttcccaag agaaggtgtt
     3721 ttcgtttcta atggtactca ctggttcgtc acacaaagaa acttctacga gccccaaatt
     3781 attactactg acaacacctt cgtatccggt aattgtgacg ttgtcatcgg tattgtcaac
     3841 aacaccgttt atgatccact ccaaccagaa ttagattcct tcaaggaaga attggataaa
     3901 tactttaaga atcacacctc ccctgacgtt gatctaggtg acatatctgg cattaacgct
     3961 tccgtggtta acattcaaaa ggaaatagac aggttaaatg aggttgccaa gaacttgaac
     4021 gagtcattga ttgatctcca agaactaggt aagtacgaac aatacattaa atggccgtgg
     4081 tacatctggt tgggttttat cgcagggctg attgctattg ttatggtgac catcatgttg
     4141 tgctgtatga cctcatgttg ttcttgttta aagggttgtt gcagctgcgg ttcttgttgc
     4201 aagttcgatg aagacgactc tgaacctgta ttgaagggtg ttaaattgca ctacacgctc
     4261 gtgccgagag gttctcacca tcatcaccat cattaaacag gccccttttc ctttgtcgat
     4321 atcatgtaat tagttatgtc acgcttacat tcacgccctc ctcccacatc cgctctaacc
     4381 gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt ttaatagtta
     4441 tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg tacaaagcgg
     4501 ccgccgaacc actagtgcac tgcagtacac ggtttccttg aaattttttt gattcggtaa
     4561 tctccgaaca gaaggaagaa cgaaggaagg agcacagact tagattggta tatatacgca
     4621 tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt aacccaactg cacagaacaa
     4681 aaacgtgcag gaaacgaaga taaatcatgt cgaaagctac atataaggaa cgtgctgcta
     4741 ctcatcctag tcctgttgct gccaagctat ttaatatcat gcacgaaaag caaacaaact
     4801 tgtgtgcttc attggatgtt cgtaccacca aggaattact ggagttagtt gaagcattag
     4861 gtcccaaaat ttgtttacta aaaacacatg tggatatctt gactgatttt tccatggagg
     4921 gcacagttaa gccgctaaag gcattatccg ccaagtacaa ttttttactc ttcgaggaca
     4981 gaaaatttgc tgacattggt aatacagtca aattgcagta ctctgcgggt gtatacagaa
     5041 tagcagaatg ggcagacatt acgaatgcac acggtgtggt gggcccaggt attgttagcg
     5101 gtttgaagca ggcggcagaa gaagtaacaa aggaacctag aggccttttg atgttagcag
     5161 aattgtcatg caagggctcc ctatctactg gagaatatac taagggtact gttgacattg
     5221 cgaagagcga caaagatttt gttatcggct ttattgctca aagagacatg ggtggaagag
     5281 atgaaggtta cgattggttg attatgacac ccggtgtggg tttagatgac aagggagatg
     5341 cattgggtca acagtataga accgtggatg atgtggtttc tacaggatct gacattatta
     5401 ttgttggaag aggactattt gcaaagggaa gggatgctaa ggtagagggt gaacgttaca
     5461 gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa aaaactgtat
     5521 tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa ttatatcagt
     5581 tattacccga gtaacgaagc atctgtgctt cattttgtag aacaaaaatg caacgcgaga
     5641 gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg
     5701 aaagcgctat tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa aaatgcaacg
     5761 cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa cagaaatgca
     5821 acgcgagagc gctattttac caacaaagaa tctatacttc ttttttgttc tacaaaaatg
     5881 catcccgaga gcgctatttt tctaacaaag catcttagat tacttttttt ctcctttgtg
     5941 cgctctataa tgcagtctct tgataacttt ttgcactgta ggtccgttaa ggttagaaga
     6001 aggctacttt ggtgtctatt ttctcttcca taaaaaaagc ctgactccac ttcccgcgtt
     6061 tactgattac tagcgaagct gcgggtgcat tttttcaaga taaaggcatc cccgattata
     6121 ttctataccg atgtggattg cgcatacttt gtgaacagaa agtgatagcg ttgatgattc
     6181 ttcattggtc agaaaattat gaacggtttc ttctattttg tctctatata ctacgtatag
     6241 gaaatgttta cattttcgta ttgttttcga ttcactctat gaatagttct tactacaatt
     6301 tttttgtcta aagagtaata ctagagataa acataaaaaa tgtagaggtc gagtttagat
     6361 gcaagttcaa ggagcgaaag gtggatgggt aggttatata gggatatagc acagagatat
     6421 atagcaaaga gatacttttg agcaatgttt gtggaagcgg tattcgcaat attttagtag
     6481 ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc gtcatcagag cgcttttggt
     6541 tttcaaaagc gctctgaagt tcctatactt tctagctaga gaataggaac ttcccgagcg
     6601 gccgcgtgtt acaagcttga ctccagcgta actggactgc aatcaactca ctggctcacc
     6661 ttcacgggtg ggcctttctt cggtagaaaa tcaaaggatc ttcttgagat cctttttttc
     6721 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     6781 cggatcaaga gctaccaact ctttttccga ggtaactggc ttcagcagag cgcagatacc
     6841 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     6901 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     6961 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     7021 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     7081 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     7141 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag gggtaaacgc
     7201 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcatc gatttttgtg
     7261 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcagaaa ggcccacccg
     7321 aaggtgagcc aggtgattac atttgggccc tcatcagagg ttttcaccgt catcaccgaa
     7381 acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc
     7441 tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct
     7501 gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcacttacc aatgcttaat
     7561 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     7621 cgtcgtgtag ataactacga tgcgggaggg cttaccatct ggccccagtg ctgcaatgat
     7681 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccgggag
     7741 ggccgagcgc agaagtgatc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7801 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7861 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7921 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7981 tcctccgatc gttgccagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     8041 actgcataat tctcttactg tcatgccatc cgtgagatgc ttttctgtga ctggtgagta
     8101 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     8161 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     8221 ttcttcgggg cgtaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     8281 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     8341 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     8401 actcatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag
     8461 tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa gtcaaaagcc
     8521 tccggtcgga ggcttttgac tttctgctat ggaggtcagg tatgatttaa atggt
//
