LOCUS       sequence-297544-        8623 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 8623)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 8623)
  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-m4046501
FEATURES             Location/Qualifiers
     source          1..8623
                     /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..4333
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(4378..4396)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             4747..5550
                     /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      5633..6615
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      6693..6722
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      6744..7331
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(7343..7370)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(7394..7585)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(7586..8377,8378..8446))
                     /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     8209..8228
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8447..8549)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(8550..8581)
                     /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 atgtttgttt ttcttgtttt attgccacta gtctctagtc agtgtgttaa
      481 tcttacaacc agaactcaat taccccctgc atacactaat tctttcacac gtggtgttta
      541 ttaccctgac aaagttttca gatcctcagt tttacattca actcaggact tgttcttacc
      601 tttcttttcc aatgttactt ggttccatgc tatacatgtc tctgggacca atggtactaa
      661 gaggtttgat aaccctgtcc taccatttaa tgatggtgtt tattttgctt ccactgagaa
      721 gtctaacata ataagaggct ggatttttgg tactacttta gattcgaaga cccagtccct
      781 acttattgtt aataacgcta ctaatgttgt tattaaagtc tgtgaatttc aattttgtaa
      841 tgatccattt ttgggtgttt attaccacaa aaacaacaaa agttggatgg aaagtgagtt
      901 cagagtttat tctagtgcga ataattgcac ttttgaatat gtctctcagc cttttcttat
      961 ggaccttgaa ggaaaacagg gtaatttcaa aaatcttagg gaatttgtgt ttaagaatat
     1021 tgatggttat tttaaaatat attctaagca cacgcctatt aatttagtgc gtgatctccc
     1081 tcagggtttt tcggctttag aaccattggt agatttgcca ataggtatta acatcactag
     1141 gtttcaaact ttacttgctt tacatagaag ttatttgact cctggtgatt cttcttcagg
     1201 ttggacagct ggtgctgcag cttattatgt gggttatctt caacctagga cttttctatt
     1261 aaaatataat gaaaatggaa ccattacaga tgctgtagac tgtgcacttg accctctctc
     1321 agaaacaaag tgtacgttga aatccttcac tgtagaaaaa ggaatctatc aaacttctaa
     1381 ctttagagtc caaccaacag aatctattgt tagatttcct aatattacaa acttgtgccc
     1441 ttttggtgaa gtttttaacg ccaccagatt tgcatctgtt tatgcttgga acaggaagag
     1501 aatcagcaac tgtgttgctg attattctgt cctatataat tccgcatcat tttccacttt
     1561 taagtgttat ggagtgtctc ctactaaatt aaatgatctc tgctttacta atgtctatgc
     1621 agattcattt gtaattagag gtgatgaagt cagacaaatc gctccagggc aaactggaaa
     1681 gattgctgat tataattata aattaccaga tgattttaca ggctgcgtta tagcttggaa
     1741 ttctaacaat cttgattcta aggttggtgg taattataat tacctgtata gattgtttag
     1801 gaagtctaat ctcaaacctt ttgagagaga tatttcaact gaaatctatc aggccggtag
     1861 cacaccttgt aatggtgttg aaggttttaa ttgttacttt cctttacaat catatggttt
     1921 ccaacccact aatggtgttg gttaccaacc atacagagta gtagtacttt cttttgaact
     1981 tctacatgca ccagcaactg tttgtggacc taaaaagtct actaatttgg ttaaaaacaa
     2041 atgtgtcaat ttcaacttca atggtttaac aggcacaggt gttcttactg agtctaacaa
     2101 aaagtttctg cctttccaac aatttggcag agacattgct gacactactg atgctgtccg
     2161 tgatccacag acacttgaga ttcttgacat tacaccatgt tcttttggtg gtgtcagtgt
     2221 tataacacca ggaacaaata cttctaacca ggttgctgtt ctttatcagg atgttaactg
     2281 cacagaagtc cctgttgcta ttcatgcaga tcaacttact cctacttggc gtgtttattc
     2341 tacaggttct aatgtttttc aaacacgtgc aggctgttta ataggggctg aacatgtcaa
     2401 caactcatat gagtgtgaca tacccattgg tgcaggtata tgcgctagtt atcagactca
     2461 gactaattct cctcggcggg cacgtagtgt agctagtcaa tccatcattg cctacactat
     2521 gtcacttggt gcagaaaatt cagttgctta ctctaataac tctattgcca tacccacaaa
     2581 ttttactatt agtgttacca cagaaattct accagtgtct atgaccaaga catcagtaga
     2641 ttgtacaatg tacatttgtg gtgattcaac tgaatgcagc aatcttttgt tgcaatatgg
     2701 cagtttttgt acacaattaa accgtgcttt aactggaata gctgttgaac aagacaaaaa
     2761 cacccaagaa gtttttgcac aagtcaaaca aatttacaaa acaccaccaa ttaaagattt
     2821 tggtggtttt aatttttcac aaatattacc agatccatca aaaccaagca agaggtcatt
     2881 tattgaagat ctacttttca acaaagtgac acttgcagat gctggcttca tcaaacaata
     2941 tggtgattgc cttggtgata ttgctgctag agacctcatt tgtgcacaaa agtttaacgg
     3001 ccttactgtt ttgccacctt tgctcacaga tgaaatgatt gctcaataca cttctgcact
     3061 gttagcgggt acaatcactt ctggttggac ctttggtgca ggtgctgcat tacaaatacc
     3121 atttgctatg caaatggctt ataggtttaa tggtattgga gttacacaga atgttctcta
     3181 tgagaaccaa aaattgattg ccaaccaatt taatagtgct attggcaaaa ttcaagactc
     3241 actttcttcc acagcaagtg cacttggaaa acttcaagat gtggtcaacc aaaatgcaca
     3301 agctttaaac acgcttgtta aacaacttag ctccaatttt ggtgcaattt caagtgtttt
     3361 aaatgatatc ctttcacgtc ttgacaaagt tgaggctgaa gtgcaaattg ataggttgat
     3421 cacaggcaga cttcaaagtt tgcagacata tgtgactcaa caattaatta gagctgcaga
     3481 aatcagagct tctgctaatc ttgctgctac taaaatgtca gagtgtgtac ttggacaatc
     3541 aaaaagagtt gatttttgtg gaaagggcta tcatcttatg tccttccctc agtcagcacc
     3601 tcatggtgta gtcttcttgc atgtgactta tgtccctgca caagaaaaga acttcacaac
     3661 tgctcctgcc atttgtcatg atggaaaagc acactttcct cgtgaaggtg tctttgtttc
     3721 aaatggcaca cactggtttg taacacaaag gaatttttat gaaccacaaa tcattactac
     3781 agacaacaca tttgtgtctg gtaactgtga tgttgtaata ggaattgtca acaacacagt
     3841 ttatgatcct ttgcaacctg aattagactc attcaaggag gagttagata aatattttaa
     3901 gaatcataca tcaccagatg ttgatttagg tgacatctct ggcattaatg cttcagttgt
     3961 aaacattcaa aaagaaattg accgcctcaa tgaggttgcc aagaatttaa atgaatctct
     4021 catcgatctc caagaacttg gaaagtatga gcagtatata aaatggccat ggtacatttg
     4081 gctaggtttt atagctggct tgattgccat agtaatggtg acaattatgc tttgctgtat
     4141 gaccagttgc tgtagttgtc tcaagggctg ttgttcttgt ggatcctgct gcaaatttga
     4201 tgaagacgac tctgagccag tgctcaaagg agtcaaatta cattacacac tcgtgccgag
     4261 aggttctgat tataaagatc atgatggtga ttataaagat catgatattg attataaaga
     4321 tgatgatgat aaataaacag gccccttttc ctttgtcgat atcatgtaat tagttatgtc
     4381 acgcttacat tcacgccctc ctcccacatc cgctctaacc gaaaaggaag gagttagaca
     4441 acctgaagtc taggtcccta tttatttttt ttaatagtta tgttagtatt aagaacgtta
     4501 tttatatttc aaatttttct tttttttctg tacaaagcgg ccgccgaacc actagtgcac
     4561 tgcagtacac ggtttccttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa
     4621 cgaaggaagg agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca
     4681 tgaaattgcc cagtattctt aacccaactg cacagaacaa aaacgtgcag gaaacgaaga
     4741 taaatcatgt cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct
     4801 gccaagctat ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt
     4861 cgtaccacca aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta
     4921 aaaacacatg tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag
     4981 gcattatccg ccaagtacaa ttttttactc ttcgaggaca gaaaatttgc tgacattggt
     5041 aatacagtca aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt
     5101 acgaatgcac acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa
     5161 gaagtaacaa aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc
     5221 ctatctactg gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt
     5281 gttatcggct ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg
     5341 attatgacac ccggtgtggg tttagatgac aagggagatg cattgggtca acagtataga
     5401 accgtggatg atgtggtttc tacaggatct gacattatta ttgttggaag aggactattt
     5461 gcaaagggaa gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca
     5521 tatttgagaa gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata
     5581 ctaaactcac aaattagagc ttcaatttaa ttatatcagt tattacccga gtaacgaagc
     5641 atctgtgctt cattttgtag aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa
     5701 gaatctgagc tgcattttta cagaacagaa atgcaacgcg aaagcgctat tttaccaacg
     5761 aagaatctgt gcttcatttt tgtaaaacaa aaatgcaacg cgagagcgct aatttttcaa
     5821 acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgagagc gctattttac
     5881 caacaaagaa tctatacttc ttttttgttc tacaaaaatg catcccgaga gcgctatttt
     5941 tctaacaaag catcttagat tacttttttt ctcctttgtg cgctctataa tgcagtctct
     6001 tgataacttt ttgcactgta ggtccgttaa ggttagaaga aggctacttt ggtgtctatt
     6061 ttctcttcca taaaaaaagc ctgactccac ttcccgcgtt tactgattac tagcgaagct
     6121 gcgggtgcat tttttcaaga taaaggcatc cccgattata ttctataccg atgtggattg
     6181 cgcatacttt gtgaacagaa agtgatagcg ttgatgattc ttcattggtc agaaaattat
     6241 gaacggtttc ttctattttg tctctatata ctacgtatag gaaatgttta cattttcgta
     6301 ttgttttcga ttcactctat gaatagttct tactacaatt tttttgtcta aagagtaata
     6361 ctagagataa acataaaaaa tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag
     6421 gtggatgggt aggttatata gggatatagc acagagatat atagcaaaga gatacttttg
     6481 agcaatgttt gtggaagcgg tattcgcaat attttagtag ctcgttacag tccggtgcgt
     6541 ttttggtttt ttgaaagtgc gtcatcagag cgcttttggt tttcaaaagc gctctgaagt
     6601 tcctatactt tctagctaga gaataggaac ttcccgagcg gccgcgtgtt acaagcttga
     6661 ctccagcgta actggactgc aatcaactca ctggctcacc ttcacgggtg ggcctttctt
     6721 cggtagaaaa tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6781 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     6841 ctttttccga ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     6901 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     6961 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     7021 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     7081 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     7141 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     7201 gaacaggaga gcgcacgagg gagcttccag gggtaaacgc ctggtatctt tatagtcctg
     7261 tcgggtttcg ccacctctga cttgagcatc gatttttgtg atgctcgtca ggggggcgga
     7321 gcctatggaa aaacgccagc aacgcagaaa ggcccacccg aaggtgagcc aggtgattac
     7381 atttgggccc tcatcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt
     7441 aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca
     7501 gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa
     7561 gggcggtttt ttcctgtttg gtcacttacc aatgcttaat cagtgaggca cctatctcag
     7621 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     7681 tgcgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     7741 cggctccaga tttatcagca ataaaccagc cagccgggag ggccgagcgc agaagtgatc
     7801 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     7861 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     7921 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     7981 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgccagaa
     8041 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     8101 tcatgccatc cgtgagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     8161 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     8221 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgtaaactct
     8281 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     8341 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     8401 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatttta gcttccttag
     8461 ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt
     8521 gaaagttgga acctcttacg tgccgatcaa gtcaaaagcc tccggtcgga ggcttttgac
     8581 tttctgctat ggaggtcagg tatgatttaa atggtcagtt aga
//
