LOCUS       sequence-300422-        8622 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 8622)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 8622)
  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-m4046299
FEATURES             Location/Qualifiers
     source          1..8622
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(331..1134)
                     /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     1485..1503
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(1548..1565)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(1566..1583)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     promoter        complement(5414..5855)
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     primer_bind     complement(5420..5443)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    5738..5855
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     terminator      5922..5953
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        5954..6056
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(6057..6125,6126..6917)
                     /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(6275..6294)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             6918..7109
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      7133..7160
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(7172..7759)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(7781..7810)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      join(7888..8622,1..248)
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 gtaaaatagc gctctcgcgt tgcatttctg ttctgtaaaa atgcagctca gattctttgt
       61 ttgaaaaatt agcgctctcg cgttgcattt ttgttttaca aaaatgaagc acagattctt
      121 cgttggtaaa atagcgcttt cgcgttgcat ttctgttctg taaaaatgca gctcagattc
      181 tttgtttgaa aaattagcgc tctcgcgttg catttttgtt ctacaaaatg aagcacagat
      241 gcttcgttac tcgggtaata actgatataa ttaaattgaa gctctaattt gtgagtttag
      301 tatacatgca tttacttata atacagtttt ttagttttgc tggccgcatc ttctcaaata
      361 tgcttcccag cctgcttttc tgtaacgttc accctctacc ttagcatccc ttccctttgc
      421 aaatagtcct cttccaacaa taataatgtc agatcctgta gaaaccacat catccacggt
      481 tctatactgt tgacccaatg catctccctt gtcatctaaa cccacaccgg gtgtcataat
      541 caaccaatcg taaccttcat ctcttccacc catgtctctt tgagcaataa agccgataac
      601 aaaatctttg tcgctcttcg caatgtcaac agtaccctta gtatattctc cagtagatag
      661 ggagcccttg catgacaatt ctgctaacat caaaaggcct ctaggttcct ttgttacttc
      721 ttctgccgcc tgcttcaaac cgctaacaat acctgggccc accacaccgt gtgcattcgt
      781 aatgtctgcc cattctgcta ttctgtatac acccgcagag tactgcaatt tgactgtatt
      841 accaatgtca gcaaattttc tgtcctcgaa gagtaaaaaa ttgtacttgg cggataatgc
      901 ctttagcggc ttaactgtgc cctccatgga aaaatcagtc aagatatcca catgtgtttt
      961 tagtaaacaa attttgggac ctaatgcttc aactaactcc agtaattcct tggtggtacg
     1021 aacatccaat gaagcacaca agtttgtttg cttttcgtgc atgatattaa atagcttggc
     1081 agcaacagga ctaggatgag tagcagcacg ttccttatat gtagctttcg acatgattta
     1141 tcttcgtttc ctgcacgttt ttgttctgtg cagttgggtt aagaatactg ggcaatttca
     1201 tgtttcttca acactacata tgcgtatata taccaatcta agtctgtgct ccttccttcg
     1261 ttcttccttc tgttcggaga ttaccgaatc aaaaaaattt caaggaaacc gtgtactgca
     1321 gtgcactagt ggttcggcgg ccgctttgta cagaaaaaaa agaaaaattt gaaatataaa
     1381 taacgttctt aatactaaca taactattaa aaaaaataaa tagggaccta gacttcaggt
     1441 tgtctaactc cttccttttc ggttagagcg gatgtgggag gagggcgtga atgtaagcgt
     1501 gacataacta attacatgat atcgacaaag gaaaaggggc ctgtttaatg atggtgatga
     1561 tggtgagaac ctctcggcac gagcgtgtag tgcaatttaa cacccttcaa tacaggttca
     1621 gagtcgtctt catcgaactt gcaacaagaa ccgcagctgc aacaaccctt taaacaagaa
     1681 caacatgagg tcatacagca caacatgatg gtcaccataa caatagcaat cagccctgcg
     1741 ataaaaccca accagatgta ccacggccat ttaatgtatt gttcgtactt acctagttct
     1801 tggagatcaa tcaatgactc gttcaagttc ttggcaacct catttaacct gtctatttcc
     1861 ttttgaatgt taaccacgga agcgttaatg ccagatatgt cacctagatc aacgtcaggg
     1921 gaggtgtgat tcttaaagta tttatccaat tcttccttga aggaatctaa ttctggttgg
     1981 agtggatcat aaacggtgtt gttgacaata ccgatgacaa cgtcacaatt accggatacg
     2041 aaggtgttgt cagtagtaat aatttggggc tcgtagaagt ttctttgtgt gacgaaccag
     2101 tgagtaccat tagaaacgaa aacaccttct cttgggaagt gagctttacc atcgtgacag
     2161 atggcaggag ctgtagtgaa gttcttttcc tgagccggca cgtaagtaac atgcaaaaag
     2221 acgacaccgt gcggggcaga ttggggaaag gacatcaaat gataaccttt accacagaag
     2281 tcaacacgct tagactgacc taggacacat tcactcatct tggtcgcggc taaattagca
     2341 ctagctctaa tttcagcagc gcggataagt tgttgagtaa cgtaggtttg tagcgattgc
     2401 aatcttccgg tgatcaatct atcgatttgt acttcggcct caaccttatc caacctagat
     2461 aagatatcgt tgagtacgga agaaatggcc ccgaagttag aggacaactg ttttacgagg
     2521 gtgttcaaag cttgagcatt ctggttaaca acgtcttgta gtttaccaag cgccgaagca
     2581 gtcgaggaca aagagtcttg aatcttacca atggccgagt tgaattgatt agcgattagc
     2641 ttttggtttt catacagaac gttttgagta actcctatcc cgttgaatct gtaagccatt
     2701 tgcatagcaa atgggatttg caatgcagca ccagcaccaa atgtccaacc agaagtaatc
     2761 gtcccagcca agagcgctga ggtatattga gcgatcattt catcagtcaa tagtgggggc
     2821 aacacggtaa gaccattgaa cttctgcgca catatcaagt ctcgggctgc aatgtcaccc
     2881 aagcaatcgc catattgctt gatgaatcca gcgtcggcta acgtaacttt attaaataat
     2941 aaatcttcga taaacgatct cttggatggc tttgatggat ctggcaagat ttggctaaag
     3001 ttgaaaccgc cgaaatcctt aatgggtggg gtcttgtaga tttgcttgac ttgggcaaat
     3061 acttcctggg tgtttttgtc ttgttcgaca gcaatacccg taagagcacg gttcaattgt
     3121 gtgcagaagg aaccgtattg taacaacaaa ttagaacact cagtagagtc accacagata
     3181 tacattgtac aatccactga ggtctttgtc atagagacag gtaagatttc agtagtaaca
     3241 gaaatagtga aattagttgg tatagcaata gaattgttac tgtatgcgac tgagttttcg
     3301 gcacctaggc tcatggtgta tgcaatgata ctttgtgaag caacacttct agctcttctt
     3361 ggtgaattag tctgtgtttg gtaggatgca cagattccgg caccaattgg aatgtcacat
     3421 tcataggagt tgttgacatg ctcagcacct atcaaacaac cagctcttgt ttggaaaaca
     3481 ttagaaccag tagagtagac tctccaagta ggggttagtt ggtcagcgtg gatagcaact
     3541 ggcacttccg tacagttgac gtcttggtat aaaacggcga cctgattaga agtgttggtt
     3601 ccaggggtaa taacgctaac accaccaaag gaacatggag tgatgtccaa aatttccaaa
     3661 gtttgaggat cccggaccgc atcggtcgta tctgcaatat ctctaccgaa ttgttggaat
     3721 ggcaggaact tcttgttgga ttcggtcaat acgccagtac cagtaagacc attgaaattg
     3781 aagttgacac acttattttt aaccaagttc gtggactttt tgggaccgca gactgtagct
     3841 ggggcatgca atagttcaaa agacaggaca acaactctat atggttggta accaactccg
     3901 ttggttggct gaaaaccgta agattgtaag gggaagtaac agttaaaacc ttcgacccca
     3961 ttacaaggag tagaaccagc ttgataaatc tcggtactta tgtctctttc gaaaggtttc
     4021 aaattagact tacgaaatag gcggtacaaa tagttatagt tcccaccaac cttcgaatcc
     4081 aagttgttgc tattccaagc gatcacgcaa ccggtgaagt cgtctggcaa tttgtagttg
     4141 taatcagcaa tcttaccggt ttgacctggc gcgatttgtc ttacttcatc gcctctaatg
     4201 acaaaggagt ctgcgtaaac attagtgaag cataagtcgt taagcttggt tggcgaaacc
     4261 ccgtaacatt tgaaagtaga gaaggaagca ctgttatata gtacagagta atctgccaca
     4321 caattggaga ttctcttacg gttccaagcg tacacagaag caaaccttgt ggcattaaaa
     4381 acctcaccga atggacacaa gttggtgata tttgggaatc tgactattga ttcagttggt
     4441 tgaactctaa agttgctagt ttggtatata cccttttcaa ctgtgaaaga ttttagggtg
     4501 cacttagttt cgctcaatgg atccaaagca cagtctacag catcggtaat tgtcccgttt
     4561 tcattgtact tgagcagaaa ggttcttggc tgcaaataac cgacatagta ggcagcagca
     4621 cccgcagtcc atccagacga ggagtcacca ggagttaaat atgatcggtg taaggctagc
     4681 aaagtttgaa atctcgtaat gttgatacca atcggcaaat caacgagtgg ttccaaagca
     4741 gagaaaccct gaggtaaatc tctaactagg ttaattggtg tgtgcttaga gtagatctta
     4801 aagtaaccgt cgatattttt gaagacaaat tctctcaagt tcttgaagtt gccttgctta
     4861 ccctccaaat ccatcaggaa aggttgactg acgtattcga aggtacagtt gttagcactt
     4921 gaataaaccc gaaattcgga ttccatccag cttttattat tcttatggta ataaacaccc
     4981 aaaaacgggt cgttacagaa ttggaactca cagactttaa ttacaacatt agtagcgttg
     5041 ttgacaatta acaaagattg agtcttggaa tcaagcgtgg ttccaaagat ccaacctcta
     5101 atgatgttag acttttctgt ggaagcgaag tagacaccgt cattgaatgg caacactggg
     5161 ttatcgaacc tcttagtacc attggtacca gaaacgtgta tggcgtgaaa ccaagtaacg
     5221 tttgaaaaga atggcaagaa caaatcttgg gtactatgta aaaccgaaga acgaaatact
     5281 ttgtcggggt aatagacgcc tctggtgaaa gagttagtgt atgctggagg tagctgcgtg
     5341 cgagttgtca gattaacgca ttgggagctc accaatggaa gcaaaactaa aaagacgaac
     5401 attatagttt tttctccttg acgttaaagt atagaggtat attaacaatt ttttgttgat
     5461 acttttatga catttgaata agaagtaata caaactgaaa atgttgaaag tattagttaa
     5521 agtggttatg cagcttttcc atttatatat ctgttaatag atcaaaaatc atcgcttcgc
     5581 tgattaatta ccccagaaat aaggctaaaa aactaatcgc attatcatcc tatggttgtt
     5641 aatttgattc gttaatttga aggtttgtgg ggccaggtta ctgccaattt ttcctcttca
     5701 taaccataaa agctagtatt gtagaatctt tattgttcgg agcagtgcgg cgcgaggcac
     5761 atctgcgttt caggaacgcg accggtgaag acgaggacgc acggaggaga gtcttccgtc
     5821 ggagggctgt cgcccgctcg gcggcttcta atccgtactt caatatagca agcggccgct
     5881 ctaactgacc atttaaatca tacctgacct ccatagcaga aagtcaaaag cctccgaccg
     5941 gaggcttttg acttgatcgg cacgtaagag gttccaactt tcaccataat gaaataagat
     6001 cactaccggg cgtatttttt gagttatcga gattttcagg agctaaggaa gctaaaatga
     6061 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     6121 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     6181 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagttta cgccccgaag
     6241 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     6301 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     6361 agtactcacc agtcacagaa aagcatctca cggatggcat gacagtaaga gaattatgca
     6421 gtgctgccat aaccatgagt gataacactg cggccaactt acttctggca acgatcggag
     6481 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     6541 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     6601 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     6661 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggatcactt ctgcgctcgg
     6721 ccctcccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     6781 gtatcattgc agcactgggg ccagatggta agccctcccg catcgtagtt atctacacga
     6841 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     6901 tgattaagca ttggtaagtg accaaacagg aaaaaaccgc ccttaacatg gcccgcttta
     6961 tcagaagcca gacattaacg cttctggaga aactcaacga gctggacgcg gatgaacagg
     7021 cagacatctg tgaatcgctt cacgaccacg ctgatgagct ttaccgcagc tgcctcgcgc
     7081 gtttcggtga tgacggtgaa aacctctgat gagggcccaa atgtaatcac ctggctcacc
     7141 ttcgggtggg cctttctgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     7201 atcacaaaaa tcgatgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     7261 aggcgtttac ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     7321 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     7381 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     7441 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     7501 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     7561 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     7621 ttggtatctg cgctctgctg aagccagtta cctcggaaaa agagttggta gctcttgatc
     7681 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     7741 cagaaaaaaa ggatctcaag aagatccttt gattttctac cgaagaaagg cccacccgtg
     7801 aaggtgagcc agtgagttga ttgcagtcca gttacgctgg agtcaagctt gtaacacgcg
     7861 gccgctcggg aagttcctat tctctagcta gaaagtatag gaacttcaga gcgcttttga
     7921 aaaccaaaag cgctctgatg acgcactttc aaaaaaccaa aaacgcaccg gactgtaacg
     7981 agctactaaa atattgcgaa taccgcttcc acaaacattg ctcaaaagta tctctttgct
     8041 atatatctct gtgctatatc cctatataac ctacccatcc acctttcgct ccttgaactt
     8101 gcatctaaac tcgacctcta cattttttat gtttatctct agtattactc tttagacaaa
     8161 aaaattgtag taagaactat tcatagagtg aatcgaaaac aatacgaaaa tgtaaacatt
     8221 tcctatacgt agtatataga gacaaaatag aagaaaccgt tcataatttt ctgaccaatg
     8281 aagaatcatc aacgctatca ctttctgttc acaaagtatg cgcaatccac atcggtatag
     8341 aatataatcg gggatgcctt tatcttgaaa aaatgcaccc gcagcttcgc tagtaatcag
     8401 taaacgcggg aagtggagtc aggctttttt tatggaagag aaaatagaca ccaaagtagc
     8461 cttcttctaa ccttaacgga cctacagtgc aaaaagttat caagagactg cattatagag
     8521 cgcacaaagg agaaaaaaag taatctaaga tgctttgtta gaaaaatagc gctctcggga
     8581 tgcatttttg tagaacaaaa aagaagtata gattctttgt tg
//
