LOCUS       sequence-296261-        8623 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 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-m4046286
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 atgttcgtct ttttagtttt gcttccattg gtgagctccc aatgcgttaa
      481 tctgacaact cgcacgcagc tacctccagc atacactaac tctttcacca gaggcgtcta
      541 ttaccccgac aaagtatttc gttcttcggt tttacatagt acccaagatt tgttcttgcc
      601 attcttttca aacgttactt ggtttcacgc catacacgtt tctggtacca atggtactaa
      661 gaggttcgat aacccagtgt tgccattcaa tgacggtgtc tacttcgctt ccacagaaaa
      721 gtctaacatc attagaggtt ggatctttgg aaccacgctt gattccaaga ctcaatcttt
      781 gttaattgtc aacaacgcta ctaatgttgt aattaaagtc tgtgagttcc aattctgtaa
      841 cgacccgttt ttgggtgttt attaccataa gaataataaa agctggatgg aatccgaatt
      901 tcgggtttat tcaagtgcta acaactgtac cttcgaatac gtcagtcaac ctttcctgat
      961 ggatttggag ggtaagcaag gcaacttcaa gaacttgaga gaatttgtct tcaaaaatat
     1021 cgacggttac tttaagatct actctaagca cacaccaatt aacctagtta gagatttacc
     1081 tcagggtttc tctgctttgg aaccactcgt tgatttgccg attggtatca acattacgag
     1141 atttcaaact ttgctagcct tacaccgatc atatttaact cctggtgact cctcgtctgg
     1201 atggactgcg ggtgctgctg cctactatgt cggttatttg cagccaagaa cctttctgct
     1261 caagtacaat gaaaacggga caattaccga tgctgtagac tgtgctttgg atccattgag
     1321 cgaaactaag tgcaccctaa aatctttcac agttgaaaag ggtatatacc aaactagcaa
     1381 ctttagagtt caaccaactg aatcaatagt cagattccca aatatcacca acttgtgtcc
     1441 attcggtgag gtttttaatg ccacaaggtt tgcttctgtg tacgcttgga accgtaagag
     1501 aatctccaat tgtgtggcag attactctgt actatataac agtgcttcct tctctacttt
     1561 caaatgttac ggggtttcgc caaccaagct taacgactta tgcttcacta atgtttacgc
     1621 agactccttt gtcattagag gcgatgaagt aagacaaatc gcgccaggtc aaaccggtaa
     1681 gattgctgat tacaactaca aattgccaga cgacttcacc ggttgcgtga tcgcttggaa
     1741 tagcaacaac ttggattcga aggttggtgg gaactataac tatttgtacc gcctatttcg
     1801 taagtctaat ttgaaacctt tcgaaagaga cataagtacc gagatttatc aagctggttc
     1861 tactccttgt aatggggtcg aaggttttaa ctgttacttc cccttacaat cttacggttt
     1921 tcagccaacc aacggagttg gttaccaacc atatagagtt gttgtcctgt cttttgaact
     1981 attgcatgcc ccagctacag tctgcggtcc caaaaagtcc acgaacttgg ttaaaaataa
     2041 gtgtgtcaac ttcaatttca atggtcttac tggtactggc gtattgaccg aatccaacaa
     2101 gaagttcctg ccattccaac aattcggtag agatattgca gatacgaccg atgcggtccg
     2161 ggatcctcaa actttggaaa ttttggacat cactccatgt tcctttggtg gtgttagcgt
     2221 tattacccct ggaaccaaca cttctaatca ggtcgccgtt ttataccaag acgtcaactg
     2281 tacggaagtg ccagttgcta tccacgctga ccaactaacc cctacttgga gagtctactc
     2341 tactggttct aatgttttcc aaacaagagc tggttgtttg ataggtgctg agcatgtcaa
     2401 caactcctat gaatgtgaca ttccaattgg tgccggaatc tgtgcatcct accaaacaca
     2461 gactaattca ccaagaagag ctagaagtgt tgcttcacaa agtatcattg catacaccat
     2521 gagcctaggt gccgaaaact cagtcgcata cagtaacaat tctattgcta taccaactaa
     2581 tttcactatt tctgttacta ctgaaatctt acctgtctct atgacaaaga cctcagtgga
     2641 ttgtacaatg tatatctgtg gtgactctac tgagtgttct aatttgttgt tacaatacgg
     2701 ttccttctgc acacaattga accgtgctct tacgggtatt gctgtcgaac aagacaaaaa
     2761 cacccaggaa gtatttgccc aagtcaagca aatctacaag accccaccca ttaaggattt
     2821 cggcggtttc aactttagcc aaatcttgcc agatccatca aagccatcca agagatcgtt
     2881 tatcgaagat ttattattta ataaagttac gttagccgac gctggattca tcaagcaata
     2941 tggcgattgc ttgggtgaca ttgcagcccg agacttgata tgtgcgcaga agttcaatgg
     3001 tcttaccgtg ttgcccccac tattgactga tgaaatgatc gctcaatata cctcagcgct
     3061 cttggctggg acgattactt ctggttggac atttggtgct ggtgctgcat tgcaaatccc
     3121 atttgctatg caaatggctt acagattcaa cgggatagga gttactcaaa acgttctgta
     3181 tgaaaaccaa aagctaatcg ctaatcaatt caactcggcc attggtaaga ttcaagactc
     3241 tttgtcctcg actgcttcgg cgcttggtaa actacaagac gttgttaacc agaatgctca
     3301 agctttgaac accctcgtaa aacagttgtc ctctaacttc ggggccattt cttccgtact
     3361 caacgatatc ttatctaggt tggataaggt tgaggccgaa gtacaaatcg atagattgat
     3421 caccggaaga ttgcaatcgc tacaaaccta cgttactcaa caacttatcc gcgctgctga
     3481 aattagagct agtgctaatt tagccgcgac caagatgagt gaatgtgtcc taggtcagtc
     3541 taagcgtgtt gacttctgtg gtaaaggtta tcatttgatg tcctttcccc aatctgcccc
     3601 gcacggtgtc gtctttttgc atgttactta cgtgccggct caggaaaaga acttcactac
     3661 agctcctgcc atctgtcacg atggtaaagc tcacttccca agagaaggtg ttttcgtttc
     3721 taatggtact cactggttcg tcacacaaag aaacttctac gagccccaaa ttattactac
     3781 tgacaacacc ttcgtatccg gtaattgtga cgttgtcatc ggtattgtca acaacaccgt
     3841 ttatgatcca ctccaaccag aattagattc cttcaaggaa gaattggata aatactttaa
     3901 gaatcacacc tcccctgacg ttgatctagg tgacatatct ggcattaacg cttccgtggt
     3961 taacattcaa aaggaaatag acaggttaaa tgaggttgcc aagaacttga acgagtcatt
     4021 gattgatctc caagaactag gtaagtacga acaatacatt aaatggccgt ggtacatctg
     4081 gttgggtttt atcgcagggc tgattgctat tgttatggtg accatcatgt tgtgctgtat
     4141 gacctcatgt tgttcttgtt taaagggttg ttgcagctgc ggttcttgtt gcaagttcga
     4201 tgaagacgac tctgaacctg tattgaaggg tgttaaattg cactacacgc 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
//
