LOCUS       sequence-296274-        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-m4046538
FEATURES             Location/Qualifiers
     source          1..8575
                     /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..4285
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4330..4348)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             4699..5502
                     /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      5585..6567
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      6645..6674
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      6696..7283
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(7295..7322)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(7346..7537)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(7538..8329,8330..8398))
                     /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     8161..8180
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8399..8501)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(8502..8533)
                     /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 aggttctcac catcatcacc atcattaaac aggccccttt tcctttgtcg atatcatgta
     4321 attagttatg tcacgcttac attcacgccc tcctcccaca tccgctctaa ccgaaaagga
     4381 aggagttaga caacctgaag tctaggtccc tatttatttt ttttaatagt tatgttagta
     4441 ttaagaacgt tatttatatt tcaaattttt cttttttttc tgtacaaagc ggccgccgaa
     4501 ccactagtgc actgcagtac acggtttcct tgaaattttt ttgattcggt aatctccgaa
     4561 cagaaggaag aacgaaggaa ggagcacaga cttagattgg tatatatacg catatgtagt
     4621 gttgaagaaa catgaaattg cccagtattc ttaacccaac tgcacagaac aaaaacgtgc
     4681 aggaaacgaa gataaatcat gtcgaaagct acatataagg aacgtgctgc tactcatcct
     4741 agtcctgttg ctgccaagct atttaatatc atgcacgaaa agcaaacaaa cttgtgtgct
     4801 tcattggatg ttcgtaccac caaggaatta ctggagttag ttgaagcatt aggtcccaaa
     4861 atttgtttac taaaaacaca tgtggatatc ttgactgatt tttccatgga gggcacagtt
     4921 aagccgctaa aggcattatc cgccaagtac aattttttac tcttcgagga cagaaaattt
     4981 gctgacattg gtaatacagt caaattgcag tactctgcgg gtgtatacag aatagcagaa
     5041 tgggcagaca ttacgaatgc acacggtgtg gtgggcccag gtattgttag cggtttgaag
     5101 caggcggcag aagaagtaac aaaggaacct agaggccttt tgatgttagc agaattgtca
     5161 tgcaagggct ccctatctac tggagaatat actaagggta ctgttgacat tgcgaagagc
     5221 gacaaagatt ttgttatcgg ctttattgct caaagagaca tgggtggaag agatgaaggt
     5281 tacgattggt tgattatgac acccggtgtg ggtttagatg acaagggaga tgcattgggt
     5341 caacagtata gaaccgtgga tgatgtggtt tctacaggat ctgacattat tattgttgga
     5401 agaggactat ttgcaaaggg aagggatgct aaggtagagg gtgaacgtta cagaaaagca
     5461 ggctgggaag catatttgag aagatgcggc cagcaaaact aaaaaactgt attataagta
     5521 aatgcatgta tactaaactc acaaattaga gcttcaattt aattatatca gttattaccc
     5581 gagtaacgaa gcatctgtgc ttcattttgt agaacaaaaa tgcaacgcga gagcgctaat
     5641 ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg cgaaagcgct
     5701 attttaccaa cgaagaatct gtgcttcatt tttgtaaaac aaaaatgcaa cgcgagagcg
     5761 ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg caacgcgaga
     5821 gcgctatttt accaacaaag aatctatact tcttttttgt tctacaaaaa tgcatcccga
     5881 gagcgctatt tttctaacaa agcatcttag attacttttt ttctcctttg tgcgctctat
     5941 aatgcagtct cttgataact ttttgcactg taggtccgtt aaggttagaa gaaggctact
     6001 ttggtgtcta ttttctcttc cataaaaaaa gcctgactcc acttcccgcg tttactgatt
     6061 actagcgaag ctgcgggtgc attttttcaa gataaaggca tccccgatta tattctatac
     6121 cgatgtggat tgcgcatact ttgtgaacag aaagtgatag cgttgatgat tcttcattgg
     6181 tcagaaaatt atgaacggtt tcttctattt tgtctctata tactacgtat aggaaatgtt
     6241 tacattttcg tattgttttc gattcactct atgaatagtt cttactacaa tttttttgtc
     6301 taaagagtaa tactagagat aaacataaaa aatgtagagg tcgagtttag atgcaagttc
     6361 aaggagcgaa aggtggatgg gtaggttata tagggatata gcacagagat atatagcaaa
     6421 gagatacttt tgagcaatgt ttgtggaagc ggtattcgca atattttagt agctcgttac
     6481 agtccggtgc gtttttggtt ttttgaaagt gcgtcatcag agcgcttttg gttttcaaaa
     6541 gcgctctgaa gttcctatac tttctagcta gagaatagga acttcccgag cggccgcgtg
     6601 ttacaagctt gactccagcg taactggact gcaatcaact cactggctca ccttcacggg
     6661 tgggcctttc ttcggtagaa aatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6721 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6781 gagctaccaa ctctttttcc gaggtaactg gcttcagcag agcgcagata ccaaatactg
     6841 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6901 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6961 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     7021 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     7081 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     7141 gcggcagggt cggaacagga gagcgcacga gggagcttcc aggggtaaac gcctggtatc
     7201 tttatagtcc tgtcgggttt cgccacctct gacttgagca tcgatttttg tgatgctcgt
     7261 caggggggcg gagcctatgg aaaaacgcca gcaacgcaga aaggcccacc cgaaggtgag
     7321 ccaggtgatt acatttgggc cctcatcaga ggttttcacc gtcatcaccg aaacgcgcga
     7381 ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt
     7441 catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc
     7501 gggccatgtt aagggcggtt ttttcctgtt tggtcactta ccaatgctta atcagtgagg
     7561 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     7621 agataactac gatgcgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     7681 acccacgctc accggctcca gatttatcag caataaacca gccagccggg agggccgagc
     7741 gcagaagtga tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     7801 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     7861 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     7921 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     7981 tcgttgccag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     8041 attctcttac tgtcatgcca tccgtgagat gcttttctgt gactggtgag tactcaacca
     8101 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     8161 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     8221 ggcgtaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     8281 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     8341 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcattt
     8401 tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta
     8461 tttcattatg gtgaaagttg gaacctctta cgtgccgatc aagtcaaaag cctccggtcg
     8521 gaggcttttg actttctgct atggaggtca ggtatgattt aaatggtcag ttaga
//
