LOCUS       sequence-300446-        8539 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 8539)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 8539)
  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-m4046640
FEATURES             Location/Qualifiers
     source          1..8539
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(325..1128)
                     /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"
     primer_bind     1479..1497
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        complement(5374..5787)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      5833..5864
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        5865..5967
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(5968..6036,6037..6828)
                     /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     complement(6186..6205)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             6829..7020
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      7044..7071
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(7083..7670)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(7692..7721)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      join(7799..8539,1..242)
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 tagcgctctc gcgttgcatt tctgttctgt aaaaatgcag ctcagattct ttgtttgaaa
       61 aattagcgct ctcgcgttgc atttttgttt tacaaaaatg aagcacagat tcttcgttgg
      121 taaaatagcg ctttcgcgtt gcatttctgt tctgtaaaaa tgcagctcag attctttgtt
      181 tgaaaaatta gcgctctcgc gttgcatttt tgttctacaa aatgaagcac agatgcttcg
      241 ttactcgggt aataactgat ataattaaat tgaagctcta atttgtgagt ttagtataca
      301 tgcatttact tataatacag ttttttagtt ttgctggccg catcttctca aatatgcttc
      361 ccagcctgct tttctgtaac gttcaccctc taccttagca tcccttccct ttgcaaatag
      421 tcctcttcca acaataataa tgtcagatcc tgtagaaacc acatcatcca cggttctata
      481 ctgttgaccc aatgcatctc ccttgtcatc taaacccaca ccgggtgtca taatcaacca
      541 atcgtaacct tcatctcttc cacccatgtc tctttgagca ataaagccga taacaaaatc
      601 tttgtcgctc ttcgcaatgt caacagtacc cttagtatat tctccagtag atagggagcc
      661 cttgcatgac aattctgcta acatcaaaag gcctctaggt tcctttgtta cttcttctgc
      721 cgcctgcttc aaaccgctaa caatacctgg gcccaccaca ccgtgtgcat tcgtaatgtc
      781 tgcccattct gctattctgt atacacccgc agagtactgc aatttgactg tattaccaat
      841 gtcagcaaat tttctgtcct cgaagagtaa aaaattgtac ttggcggata atgcctttag
      901 cggcttaact gtgccctcca tggaaaaatc agtcaagata tccacatgtg tttttagtaa
      961 acaaattttg ggacctaatg cttcaactaa ctccagtaat tccttggtgg tacgaacatc
     1021 caatgaagca cacaagtttg tttgcttttc gtgcatgata ttaaatagct tggcagcaac
     1081 aggactagga tgagtagcag cacgttcctt atatgtagct ttcgacatga tttatcttcg
     1141 tttcctgcac gtttttgttc tgtgcagttg ggttaagaat actgggcaat ttcatgtttc
     1201 ttcaacacta catatgcgta tatataccaa tctaagtctg tgctccttcc ttcgttcttc
     1261 cttctgttcg gagattaccg aatcaaaaaa atttcaagga aaccgtgtac tgcagtgcac
     1321 tagtggttcg gcggccgctt tgtacagaaa aaaaagaaaa atttgaaata taaataacgt
     1381 tcttaatact aacataacta ttaaaaaaaa taaataggga cctagacttc aggttgtcta
     1441 actccttcct tttcggttag agcggatgtg ggaggagggc gtgaatgtaa gcgtgacata
     1501 actaattaca tgatatcgac aaaggaaaag gggcctgttt acgtgtagtg caatttaaca
     1561 cccttcaata caggttcaga gtcgtcttca tcgaacttgc aacaagaacc gcagctgcaa
     1621 caacccttta aacaagaaca acatgaggtc atacagcaca acatgatggt caccataaca
     1681 atagcaatca gccctgcgat aaaacccaac cagatgtacc acggccattt aatgtattgt
     1741 tcgtacttac ctagttcttg gagatcaatc aatgactcgt tcaagttctt ggcaacctca
     1801 tttaacctgt ctatttcctt ttgaatgtta accacggaag cgttaatgcc agatatgtca
     1861 cctagatcaa cgtcagggga ggtgtgattc ttaaagtatt tatccaattc ttccttgaag
     1921 gaatctaatt ctggttggag tggatcataa acggtgttgt tgacaatacc gatgacaacg
     1981 tcacaattac cggatacgaa ggtgttgtca gtagtaataa tttggggctc gtagaagttt
     2041 ctttgtgtga cgaaccagtg agtaccatta gaaacgaaaa caccttctct tgggaagtga
     2101 gctttaccat cgtgacagat ggcaggagct gtagtgaagt tcttttcctg agccggcacg
     2161 taagtaacat gcaaaaagac gacaccgtgc ggggcagatt ggggaaagga catcaaatga
     2221 taacctttac cacagaagtc aacacgctta gactgaccta ggacacattc actcatcttg
     2281 gtcgcggcta aattagcact agctctaatt tcagcagcgc ggataagttg ttgagtaacg
     2341 taggtttgta gcgattgcaa tcttccggtg atcaatctat cgatttgtac ttcggcctca
     2401 accttatcca acctagataa gatatcgttg agtacggaag aaatggcccc gaagttagag
     2461 gacaactgtt ttacgagggt gttcaaagct tgagcattct ggttaacaac gtcttgtagt
     2521 ttaccaagcg ccgaagcagt cgaggacaaa gagtcttgaa tcttaccaat ggccgagttg
     2581 aattgattag cgattagctt ttggttttca tacagaacgt tttgagtaac tcctatcccg
     2641 ttgaatctgt aagccatttg catagcaaat gggatttgca atgcagcacc agcaccaaat
     2701 gtccaaccag aagtaatcgt cccagccaag agcgctgagg tatattgagc gatcatttca
     2761 tcagtcaata gtgggggcaa cacggtaaga ccattgaact tctgcgcaca tatcaagtct
     2821 cgggctgcaa tgtcacccaa gcaatcgcca tattgcttga tgaatccagc gtcggctaac
     2881 gtaactttat taaataataa atcttcgata aacgatctct tggatggctt tgatggatct
     2941 ggcaagattt ggctaaagtt gaaaccgccg aaatccttaa tgggtggggt cttgtagatt
     3001 tgcttgactt gggcaaatac ttcctgggtg tttttgtctt gttcgacagc aatacccgta
     3061 agagcacggt tcaattgtgt gcagaaggaa ccgtattgta acaacaaatt agaacactca
     3121 gtagagtcac cacagatata cattgtacaa tccactgagg tctttgtcat agagacaggt
     3181 aagatttcag tagtaacaga aatagtgaaa ttagttggta tagcaataga attgttactg
     3241 tatgcgactg agttttcggc acctaggctc atggtgtatg caatgatact ttgtgaagca
     3301 acacttctag ctcttcttgg tgaattagtc tgtgtttggt aggatgcaca gattccggca
     3361 ccaattggaa tgtcacattc ataggagttg ttgacatgct cagcacctat caaacaacca
     3421 gctcttgttt ggaaaacatt agaaccagta gagtagactc tccaagtagg ggttagttgg
     3481 tcagcgtgga tagcaactgg cacttccgta cagttgacgt cttggtataa aacggcgacc
     3541 tgattagaag tgttggttcc aggggtaata acgctaacac caccaaagga acatggagtg
     3601 atgtccaaaa tttccaaagt ttgaggatcc cggaccgcat cggtcgtatc tgcaatatct
     3661 ctaccgaatt gttggaatgg caggaacttc ttgttggatt cggtcaatac gccagtacca
     3721 gtaagaccat tgaaattgaa gttgacacac ttatttttaa ccaagttcgt ggactttttg
     3781 ggaccgcaga ctgtagctgg ggcatgcaat agttcaaaag acaggacaac aactctatat
     3841 ggttggtaac caactccgtt ggttggctga aaaccgtaag attgtaaggg gaagtaacag
     3901 ttaaaacctt cgaccccatt acaaggagta gaaccagctt gataaatctc ggtacttatg
     3961 tctctttcga aaggtttcaa attagactta cgaaataggc ggtacaaata gttatagttc
     4021 ccaccaacct tcgaatccaa gttgttgcta ttccaagcga tcacgcaacc ggtgaagtcg
     4081 tctggcaatt tgtagttgta atcagcaatc ttaccggttt gacctggcgc gatttgtctt
     4141 acttcatcgc ctctaatgac aaaggagtct gcgtaaacat tagtgaagca taagtcgtta
     4201 agcttggttg gcgaaacccc gtaacatttg aaagtagaga aggaagcact gttatatagt
     4261 acagagtaat ctgccacaca attggagatt ctcttacggt tccaagcgta cacagaagca
     4321 aaccttgtgg cattaaaaac ctcaccgaat ggacacaagt tggtgatatt tgggaatctg
     4381 actattgatt cagttggttg aactctaaag ttgctagttt ggtatatacc cttttcaact
     4441 gtgaaagatt ttagggtgca cttagtttcg ctcaatggat ccaaagcaca gtctacagca
     4501 tcggtaattg tcccgttttc attgtacttg agcagaaagg ttcttggctg caaataaccg
     4561 acatagtagg cagcagcacc cgcagtccat ccagacgagg agtcaccagg agttaaatat
     4621 gatcggtgta aggctagcaa agtttgaaat ctcgtaatgt tgataccaat cggcaaatca
     4681 acgagtggtt ccaaagcaga gaaaccctga ggtaaatctc taactaggtt aattggtgtg
     4741 tgcttagagt agatcttaaa gtaaccgtcg atatttttga agacaaattc tctcaagttc
     4801 ttgaagttgc cttgcttacc ctccaaatcc atcaggaaag gttgactgac gtattcgaag
     4861 gtacagttgt tagcacttga ataaacccga aattcggatt ccatccagct tttattattc
     4921 ttatggtaat aaacacccaa aaacgggtcg ttacagaatt ggaactcaca gactttaatt
     4981 acaacattag tagcgttgtt gacaattaac aaagattgag tcttggaatc aagcgtggtt
     5041 ccaaagatcc aacctctaat gatgttagac ttttctgtgg aagcgaagta gacaccgtca
     5101 ttgaatggca acactgggtt atcgaacctc ttagtaccat tggtaccaga aacgtgtatg
     5161 gcgtgaaacc aagtaacgtt tgaaaagaat ggcaagaaca aatcttgggt actatgtaaa
     5221 accgaagaac gaaatacttt gtcggggtaa tagacgcctc tggtgaaaga gttagtgtat
     5281 gctggaggta gctgcgtgcg agttgtcaga ttaacgcatt gggagctcac caatggaagc
     5341 aaaactaaaa agacgaacat tttgtaatta aaacttagat tagattgcta tgctttcttt
     5401 ctaatgagca agaagtaaaa aaagttgtaa tagaacaaga aaaatgaaac tgaaacttga
     5461 gaaattgaag accgtttatt aacttaaata tcaatgggag gtcatcgaaa gagaaaaaaa
     5521 tcaaaaaaaa aaattttcaa gaaaaagaaa cgtgataaaa atttttattg cctttttcga
     5581 cgaagaaaaa gaaacgaggc ggtctctttt ttcttttcca aacctttagt acgggtaatt
     5641 aacgacaccc tagaggaaga aagaggggaa atttagtatg ctgtgcttgg gtgttttgaa
     5701 gtggtacggc gatgcgcgga gtccgagaaa atctggaaga gtaaaaaagg agtagaaaca
     5761 ttttgaagct atggtgtgtg gggcggccgc tctaactgac catttaaatc atacctgacc
     5821 tccatagcag aaagtcaaaa gcctccgacc ggaggctttt gacttgatcg gcacgtaaga
     5881 ggttccaact ttcaccataa tgaaataaga tcactaccgg gcgtattttt tgagttatcg
     5941 agattttcag gagctaagga agctaaaatg agtattcaac atttccgtgt cgcccttatt
     6001 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     6061 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     6121 ggtaagatcc ttgagagttt acgccccgaa gaacgttttc caatgatgag cacttttaaa
     6181 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     6241 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctc
     6301 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     6361 gcggccaact tacttctggc aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     6421 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     6481 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     6541 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     6601 gataaagttg caggatcact tctgcgctcg gccctcccgg ctggctggtt tattgctgat
     6661 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     6721 aagccctccc gcatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     6781 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaagt gaccaaacag
     6841 gaaaaaaccg cccttaacat ggcccgcttt atcagaagcc agacattaac gcttctggag
     6901 aaactcaacg agctggacgc ggatgaacag gcagacatct gtgaatcgct tcacgaccac
     6961 gctgatgagc tttaccgcag ctgcctcgcg cgtttcggtg atgacggtga aaacctctga
     7021 tgagggccca aatgtaatca cctggctcac cttcgggtgg gcctttctgc gttgctggcg
     7081 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgatgctc aagtcagagg
     7141 tggcgaaacc cgacaggact ataaagatac caggcgttta cccctggaag ctccctcgtg
     7201 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     7261 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     7321 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     7381 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     7441 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     7501 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     7561 acctcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     7621 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     7681 tgattttcta ccgaagaaag gcccacccgt gaaggtgagc cagtgagttg attgcagtcc
     7741 agttacgctg gagtcaagct tgtaacacgc ggccgctcgg gaagttccta ttctctagct
     7801 agaaagtata ggaacttcag agcgcttttg aaaaccaaaa gcgctctgat gacgcacttt
     7861 caaaaaacca aaaacgcacc ggactgtaac gagctactaa aatattgcga ataccgcttc
     7921 cacaaacatt gctcaaaagt atctctttgc tatatatctc tgtgctatat ccctatataa
     7981 cctacccatc cacctttcgc tccttgaact tgcatctaaa ctcgacctct acatttttta
     8041 tgtttatctc tagtattact ctttagacaa aaaaattgta gtaagaacta ttcatagagt
     8101 gaatcgaaaa caatacgaaa atgtaaacat ttcctatacg tagtatatag agacaaaata
     8161 gaagaaaccg ttcataattt tctgaccaat gaagaatcat caacgctatc actttctgtt
     8221 cacaaagtat gcgcaatcca catcggtata gaatataatc ggggatgcct ttatcttgaa
     8281 aaaatgcacc cgcagcttcg ctagtaatca gtaaacgcgg gaagtggagt caggcttttt
     8341 ttatggaaga gaaaatagac accaaagtag ccttcttcta accttaacgg acctacagtg
     8401 caaaaagtta tcaagagact gcattataga gcgcacaaag gagaaaaaaa gtaatctaag
     8461 atgctttgtt agaaaaatag cgctctcggg atgcattttt gtagaacaaa aaagaagtat
     8521 agattctttg ttggtaaaa
//
