LOCUS       sequence-304469-        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-m4046406
FEATURES             Location/Qualifiers
     source          1..8622
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      137..166
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      188..775
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(787..814)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(838..1029)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(1030..1821,1822..1890))
                     /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     1653..1672
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1891..1993)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(1994..2025)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        2092..2533
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    2092..2209
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     2504..2527
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             6364..6381
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             6382..6399
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6444..6462)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             6813..7616
                     /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      join(7699..8622,1..59)
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 gtgcgtcatc agagcgcttt tggttttcaa aagcgctctg aagttcctat actttctagc
       61 tagagaatag gaacttcccg agcggccgcg tgttacaagc ttgactccag cgtaactgga
      121 ctgcaatcaa ctcactggct caccttcacg ggtgggcctt tcttcggtag aaaatcaaag
      181 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
      241 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaggtaac
      301 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
      361 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
      421 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
      481 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
      541 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
      601 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
      661 gagggagctt ccaggggtaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
      721 ctgacttgag catcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
      781 cagcaacgca gaaaggccca cccgaaggtg agccaggtga ttacatttgg gccctcatca
      841 gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg
      901 gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc
      961 cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg
     1021 tttggtcact taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     1081 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatgcggg agggcttacc
     1141 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     1201 agcaataaac cagccagccg ggagggccga gcgcagaagt gatcctgcaa ctttatccgc
     1261 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     1321 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     1381 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     1441 caaaaaagcg gttagctcct tcggtcctcc gatcgttgcc agaagtaagt tggccgcagt
     1501 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtgag
     1561 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     1621 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     1681 aaaagtgctc atcattggaa aacgttcttc ggggcgtaaa ctctcaagga tcttaccgct
     1741 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     1801 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     1861 aagggcgaca cggaaatgtt gaatactcat tttagcttcc ttagctcctg aaaatctcga
     1921 taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt tggaacctct
     1981 tacgtgccga tcaagtcaaa agcctccggt cggaggcttt tgactttctg ctatggaggt
     2041 caggtatgat ttaaatggtc agttagagcg gccgcttgct atattgaagt acggattaga
     2101 agccgccgag cgggcgacag ccctccgacg gaagactctc ctccgtgcgt cctcgtcttc
     2161 accggtcgcg ttcctgaaac gcagatgtgc ctcgcgccgc actgctccga acaataaaga
     2221 ttctacaata ctagctttta tggttatgaa gaggaaaaat tggcagtaac ctggccccac
     2281 aaaccttcaa attaacgaat caaattaaca accataggat gataatgcga ttagtttttt
     2341 agccttattt ctggggtaat taatcagcga agcgatgatt tttgatctat taacagatat
     2401 ataaatggaa aagctgcata accactttaa ctaatacttt caacattttc agtttgtatt
     2461 acttcttatt caaatgtcat aaaagtatca acaaaaaatt gttaatatac ctctatactt
     2521 taacgtcaag gagaaaaaac tataatgttt gtttttcttg ttttattgcc actagtctct
     2581 agtcagtgtg ttaatcttac aaccagaact caattacccc ctgcatacac taattctttc
     2641 acacgtggtg tttattaccc tgacaaagtt ttcagatcct cagttttaca ttcaactcag
     2701 gacttgttct tacctttctt ttccaatgtt acttggttcc atgctataca tgtctctggg
     2761 accaatggta ctaagaggtt tgataaccct gtcctaccat ttaatgatgg tgtttatttt
     2821 gcttccactg agaagtctaa cataataaga ggctggattt ttggtactac tttagattcg
     2881 aagacccagt ccctacttat tgttaataac gctactaatg ttgttattaa agtctgtgaa
     2941 tttcaatttt gtaatgatcc atttttgggt gtttattacc acaaaaacaa caaaagttgg
     3001 atggaaagtg agttcagagt ttattctagt gcgaataatt gcacttttga atatgtctct
     3061 cagccttttc ttatggacct tgaaggaaaa cagggtaatt tcaaaaatct tagggaattt
     3121 gtgtttaaga atattgatgg ttattttaaa atatattcta agcacacgcc tattaattta
     3181 gtgcgtgatc tccctcaggg tttttcggct ttagaaccat tggtagattt gccaataggt
     3241 attaacatca ctaggtttca aactttactt gctttacata gaagttattt gactcctggt
     3301 gattcttctt caggttggac agctggtgct gcagcttatt atgtgggtta tcttcaacct
     3361 aggacttttc tattaaaata taatgaaaat ggaaccatta cagatgctgt agactgtgca
     3421 cttgaccctc tctcagaaac aaagtgtacg ttgaaatcct tcactgtaga aaaaggaatc
     3481 tatcaaactt ctaactttag agtccaacca acagaatcta ttgttagatt tcctaatatt
     3541 acaaacttgt gcccttttgg tgaagttttt aacgccacca gatttgcatc tgtttatgct
     3601 tggaacagga agagaatcag caactgtgtt gctgattatt ctgtcctata taattccgca
     3661 tcattttcca cttttaagtg ttatggagtg tctcctacta aattaaatga tctctgcttt
     3721 actaatgtct atgcagattc atttgtaatt agaggtgatg aagtcagaca aatcgctcca
     3781 gggcaaactg gaaagattgc tgattataat tataaattac cagatgattt tacaggctgc
     3841 gttatagctt ggaattctaa caatcttgat tctaaggttg gtggtaatta taattacctg
     3901 tatagattgt ttaggaagtc taatctcaaa ccttttgaga gagatatttc aactgaaatc
     3961 tatcaggccg gtagcacacc ttgtaatggt gttgaaggtt ttaattgtta ctttccttta
     4021 caatcatatg gtttccaacc cactaatggt gttggttacc aaccatacag agtagtagta
     4081 ctttcttttg aacttctaca tgcaccagca actgtttgtg gacctaaaaa gtctactaat
     4141 ttggttaaaa acaaatgtgt caatttcaac ttcaatggtt taacaggcac aggtgttctt
     4201 actgagtcta acaaaaagtt tctgcctttc caacaatttg gcagagacat tgctgacact
     4261 actgatgctg tccgtgatcc acagacactt gagattcttg acattacacc atgttctttt
     4321 ggtggtgtca gtgttataac accaggaaca aatacttcta accaggttgc tgttctttat
     4381 caggatgtta actgcacaga agtccctgtt gctattcatg cagatcaact tactcctact
     4441 tggcgtgttt attctacagg ttctaatgtt tttcaaacac gtgcaggctg tttaataggg
     4501 gctgaacatg tcaacaactc atatgagtgt gacataccca ttggtgcagg tatatgcgct
     4561 agttatcaga ctcagactaa ttctcctcgg cgggcacgta gtgtagctag tcaatccatc
     4621 attgcctaca ctatgtcact tggtgcagaa aattcagttg cttactctaa taactctatt
     4681 gccataccca caaattttac tattagtgtt accacagaaa ttctaccagt gtctatgacc
     4741 aagacatcag tagattgtac aatgtacatt tgtggtgatt caactgaatg cagcaatctt
     4801 ttgttgcaat atggcagttt ttgtacacaa ttaaaccgtg ctttaactgg aatagctgtt
     4861 gaacaagaca aaaacaccca agaagttttt gcacaagtca aacaaattta caaaacacca
     4921 ccaattaaag attttggtgg ttttaatttt tcacaaatat taccagatcc atcaaaacca
     4981 agcaagaggt catttattga agatctactt ttcaacaaag tgacacttgc agatgctggc
     5041 ttcatcaaac aatatggtga ttgccttggt gatattgctg ctagagacct catttgtgca
     5101 caaaagttta acggccttac tgttttgcca cctttgctca cagatgaaat gattgctcaa
     5161 tacacttctg cactgttagc gggtacaatc acttctggtt ggacctttgg tgcaggtgct
     5221 gcattacaaa taccatttgc tatgcaaatg gcttataggt ttaatggtat tggagttaca
     5281 cagaatgttc tctatgagaa ccaaaaattg attgccaacc aatttaatag tgctattggc
     5341 aaaattcaag actcactttc ttccacagca agtgcacttg gaaaacttca agatgtggtc
     5401 aaccaaaatg cacaagcttt aaacacgctt gttaaacaac ttagctccaa ttttggtgca
     5461 atttcaagtg ttttaaatga tatcctttca cgtcttgaca aagttgaggc tgaagtgcaa
     5521 attgataggt tgatcacagg cagacttcaa agtttgcaga catatgtgac tcaacaatta
     5581 attagagctg cagaaatcag agcttctgct aatcttgctg ctactaaaat gtcagagtgt
     5641 gtacttggac aatcaaaaag agttgatttt tgtggaaagg gctatcatct tatgtccttc
     5701 cctcagtcag cacctcatgg tgtagtcttc ttgcatgtga cttatgtccc tgcacaagaa
     5761 aagaacttca caactgctcc tgccatttgt catgatggaa aagcacactt tcctcgtgaa
     5821 ggtgtctttg tttcaaatgg cacacactgg tttgtaacac aaaggaattt ttatgaacca
     5881 caaatcatta ctacagacaa cacatttgtg tctggtaact gtgatgttgt aataggaatt
     5941 gtcaacaaca cagtttatga tcctttgcaa cctgaattag actcattcaa ggaggagtta
     6001 gataaatatt ttaagaatca tacatcacca gatgttgatt taggtgacat ctctggcatt
     6061 aatgcttcag ttgtaaacat tcaaaaagaa attgaccgcc tcaatgaggt tgccaagaat
     6121 ttaaatgaat ctctcatcga tctccaagaa cttggaaagt atgagcagta tataaaatgg
     6181 ccatggtaca tttggctagg ttttatagct ggcttgattg ccatagtaat ggtgacaatt
     6241 atgctttgct gtatgaccag ttgctgtagt tgtctcaagg gctgttgttc ttgtggatcc
     6301 tgctgcaaat ttgatgaaga cgactctgag ccagtgctca aaggagtcaa attacattac
     6361 acactcgtgc cgagaggttc tcaccatcat caccatcatt aaacaggccc cttttccttt
     6421 gtcgatatca tgtaattagt tatgtcacgc ttacattcac gccctcctcc cacatccgct
     6481 ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta ttttttttaa
     6541 tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt tttctgtaca
     6601 aagcggccgc cgaaccacta gtgcactgca gtacacggtt tccttgaaat ttttttgatt
     6661 cggtaatctc cgaacagaag gaagaacgaa ggaaggagca cagacttaga ttggtatata
     6721 tacgcatatg tagtgttgaa gaaacatgaa attgcccagt attcttaacc caactgcaca
     6781 gaacaaaaac gtgcaggaaa cgaagataaa tcatgtcgaa agctacatat aaggaacgtg
     6841 ctgctactca tcctagtcct gttgctgcca agctatttaa tatcatgcac gaaaagcaaa
     6901 caaacttgtg tgcttcattg gatgttcgta ccaccaagga attactggag ttagttgaag
     6961 cattaggtcc caaaatttgt ttactaaaaa cacatgtgga tatcttgact gatttttcca
     7021 tggagggcac agttaagccg ctaaaggcat tatccgccaa gtacaatttt ttactcttcg
     7081 aggacagaaa atttgctgac attggtaata cagtcaaatt gcagtactct gcgggtgtat
     7141 acagaatagc agaatgggca gacattacga atgcacacgg tgtggtgggc ccaggtattg
     7201 ttagcggttt gaagcaggcg gcagaagaag taacaaagga acctagaggc cttttgatgt
     7261 tagcagaatt gtcatgcaag ggctccctat ctactggaga atatactaag ggtactgttg
     7321 acattgcgaa gagcgacaaa gattttgtta tcggctttat tgctcaaaga gacatgggtg
     7381 gaagagatga aggttacgat tggttgatta tgacacccgg tgtgggttta gatgacaagg
     7441 gagatgcatt gggtcaacag tatagaaccg tggatgatgt ggtttctaca ggatctgaca
     7501 ttattattgt tggaagagga ctatttgcaa agggaaggga tgctaaggta gagggtgaac
     7561 gttacagaaa agcaggctgg gaagcatatt tgagaagatg cggccagcaa aactaaaaaa
     7621 ctgtattata agtaaatgca tgtatactaa actcacaaat tagagcttca atttaattat
     7681 atcagttatt acccgagtaa cgaagcatct gtgcttcatt ttgtagaaca aaaatgcaac
     7741 gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc
     7801 aacgcgaaag cgctatttta ccaacgaaga atctgtgctt catttttgta aaacaaaaat
     7861 gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga
     7921 aatgcaacgc gagagcgcta ttttaccaac aaagaatcta tacttctttt ttgttctaca
     7981 aaaatgcatc ccgagagcgc tatttttcta acaaagcatc ttagattact ttttttctcc
     8041 tttgtgcgct ctataatgca gtctcttgat aactttttgc actgtaggtc cgttaaggtt
     8101 agaagaaggc tactttggtg tctattttct cttccataaa aaaagcctga ctccacttcc
     8161 cgcgtttact gattactagc gaagctgcgg gtgcattttt tcaagataaa ggcatccccg
     8221 attatattct ataccgatgt ggattgcgca tactttgtga acagaaagtg atagcgttga
     8281 tgattcttca ttggtcagaa aattatgaac ggtttcttct attttgtctc tatatactac
     8341 gtataggaaa tgtttacatt ttcgtattgt tttcgattca ctctatgaat agttcttact
     8401 acaatttttt tgtctaaaga gtaatactag agataaacat aaaaaatgta gaggtcgagt
     8461 ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt tatataggga tatagcacag
     8521 agatatatag caaagagata cttttgagca atgtttgtgg aagcggtatt cgcaatattt
     8581 tagtagctcg ttacagtccg gtgcgttttt ggttttttga aa
//
