LOCUS       sequence-300404-        8623 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 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-m4046501
FEATURES             Location/Qualifiers
     source          1..8623
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1592..2005)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      2051..2082
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        2083..2185
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(2186..2254,2255..3046)
                     /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(2404..2423)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             3047..3238
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      3262..3289
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(3301..3888)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3910..3939)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4017..4999
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(5082..5885)
                     /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     6236..6254
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(6299..6364)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(6365..6382)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
ORIGIN
        1 tccacaaaca gttgctggtg catgtagaag ttcaaaagaa agtactacta ctctgtatgg
       61 ttggtaacca acaccattag tgggttggaa accatatgat tgtaaaggaa agtaacaatt
      121 aaaaccttca acaccattac aaggtgtgct accggcctga tagatttcag ttgaaatatc
      181 tctctcaaaa ggtttgagat tagacttcct aaacaatcta tacaggtaat tataattacc
      241 accaacctta gaatcaagat tgttagaatt ccaagctata acgcagcctg taaaatcatc
      301 tggtaattta taattataat cagcaatctt tccagtttgc cctggagcga tttgtctgac
      361 ttcatcacct ctaattacaa atgaatctgc atagacatta gtaaagcaga gatcatttaa
      421 tttagtagga gacactccat aacacttaaa agtggaaaat gatgcggaat tatataggac
      481 agaataatca gcaacacagt tgctgattct cttcctgttc caagcataaa cagatgcaaa
      541 tctggtggcg ttaaaaactt caccaaaagg gcacaagttt gtaatattag gaaatctaac
      601 aatagattct gttggttgga ctctaaagtt agaagtttga tagattcctt tttctacagt
      661 gaaggatttc aacgtacact ttgtttctga gagagggtca agtgcacagt ctacagcatc
      721 tgtaatggtt ccattttcat tatattttaa tagaaaagtc ctaggttgaa gataacccac
      781 ataataagct gcagcaccag ctgtccaacc tgaagaagaa tcaccaggag tcaaataact
      841 tctatgtaaa gcaagtaaag tttgaaacct agtgatgtta atacctattg gcaaatctac
      901 caatggttct aaagccgaaa aaccctgagg gagatcacgc actaaattaa taggcgtgtg
      961 cttagaatat attttaaaat aaccatcaat attcttaaac acaaattccc taagattttt
     1021 gaaattaccc tgttttcctt caaggtccat aagaaaaggc tgagagacat attcaaaagt
     1081 gcaattattc gcactagaat aaactctgaa ctcactttcc atccaacttt tgttgttttt
     1141 gtggtaataa acacccaaaa atggatcatt acaaaattga aattcacaga ctttaataac
     1201 aacattagta gcgttattaa caataagtag ggactgggtc ttcgaatcta aagtagtacc
     1261 aaaaatccag cctcttatta tgttagactt ctcagtggaa gcaaaataaa caccatcatt
     1321 aaatggtagg acagggttat caaacctctt agtaccattg gtcccagaga catgtatagc
     1381 atggaaccaa gtaacattgg aaaagaaagg taagaacaag tcctgagttg aatgtaaaac
     1441 tgaggatctg aaaactttgt cagggtaata aacaccacgt gtgaaagaat tagtgtatgc
     1501 agggggtaat tgagttctgg ttgtaagatt aacacactga ctagagacta gtggcaataa
     1561 aacaagaaaa acaaacattt tgtaattaaa acttagatta gattgctatg ctttctttct
     1621 aatgagcaag aagtaaaaaa agttgtaata gaacaagaaa aatgaaactg aaacttgaga
     1681 aattgaagac cgtttattaa cttaaatatc aatgggaggt catcgaaaga gaaaaaaatc
     1741 aaaaaaaaaa attttcaaga aaaagaaacg tgataaaaat ttttattgcc tttttcgacg
     1801 aagaaaaaga aacgaggcgg tctctttttt cttttccaaa cctttagtac gggtaattaa
     1861 cgacacccta gaggaagaaa gaggggaaat ttagtatgct gtgcttgggt gttttgaagt
     1921 ggtacggcga tgcgcggagt ccgagaaaat ctggaagagt aaaaaaggag tagaaacatt
     1981 ttgaagctat ggtgtgtggg gcggccgctc taactgacca tttaaatcat acctgacctc
     2041 catagcagaa agtcaaaagc ctccgaccgg aggcttttga cttgatcggc acgtaagagg
     2101 ttccaacttt caccataatg aaataagatc actaccgggc gtattttttg agttatcgag
     2161 attttcagga gctaaggaag ctaaaatgag tattcaacat ttccgtgtcg cccttattcc
     2221 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     2281 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     2341 taagatcctt gagagtttac gccccgaaga acgttttcca atgatgagca cttttaaagt
     2401 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     2461 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatctcac
     2521 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     2581 ggccaactta cttctggcaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     2641 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     2701 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     2761 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     2821 taaagttgca ggatcacttc tgcgctcggc cctcccggct ggctggttta ttgctgataa
     2881 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     2941 gccctcccgc atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     3001 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaagtga ccaaacagga
     3061 aaaaaccgcc cttaacatgg cccgctttat cagaagccag acattaacgc ttctggagaa
     3121 actcaacgag ctggacgcgg atgaacaggc agacatctgt gaatcgcttc acgaccacgc
     3181 tgatgagctt taccgcagct gcctcgcgcg tttcggtgat gacggtgaaa acctctgatg
     3241 agggcccaaa tgtaatcacc tggctcacct tcgggtgggc ctttctgcgt tgctggcgtt
     3301 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgatgctcaa gtcagaggtg
     3361 gcgaaacccg acaggactat aaagatacca ggcgtttacc cctggaagct ccctcgtgcg
     3421 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3481 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3541 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3601 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3661 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3721 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     3781 ctcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     3841 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3901 attttctacc gaagaaaggc ccacccgtga aggtgagcca gtgagttgat tgcagtccag
     3961 ttacgctgga gtcaagcttg taacacgcgg ccgctcggga agttcctatt ctctagctag
     4021 aaagtatagg aacttcagag cgcttttgaa aaccaaaagc gctctgatga cgcactttca
     4081 aaaaaccaaa aacgcaccgg actgtaacga gctactaaaa tattgcgaat accgcttcca
     4141 caaacattgc tcaaaagtat ctctttgcta tatatctctg tgctatatcc ctatataacc
     4201 tacccatcca cctttcgctc cttgaacttg catctaaact cgacctctac attttttatg
     4261 tttatctcta gtattactct ttagacaaaa aaattgtagt aagaactatt catagagtga
     4321 atcgaaaaca atacgaaaat gtaaacattt cctatacgta gtatatagag acaaaataga
     4381 agaaaccgtt cataattttc tgaccaatga agaatcatca acgctatcac tttctgttca
     4441 caaagtatgc gcaatccaca tcggtataga atataatcgg ggatgccttt atcttgaaaa
     4501 aatgcacccg cagcttcgct agtaatcagt aaacgcggga agtggagtca ggcttttttt
     4561 atggaagaga aaatagacac caaagtagcc ttcttctaac cttaacggac ctacagtgca
     4621 aaaagttatc aagagactgc attatagagc gcacaaagga gaaaaaaagt aatctaagat
     4681 gctttgttag aaaaatagcg ctctcgggat gcatttttgt agaacaaaaa agaagtatag
     4741 attctttgtt ggtaaaatag cgctctcgcg ttgcatttct gttctgtaaa aatgcagctc
     4801 agattctttg tttgaaaaat tagcgctctc gcgttgcatt tttgttttac aaaaatgaag
     4861 cacagattct tcgttggtaa aatagcgctt tcgcgttgca tttctgttct gtaaaaatgc
     4921 agctcagatt ctttgtttga aaaattagcg ctctcgcgtt gcatttttgt tctacaaaat
     4981 gaagcacaga tgcttcgtta ctcgggtaat aactgatata attaaattga agctctaatt
     5041 tgtgagttta gtatacatgc atttacttat aatacagttt tttagttttg ctggccgcat
     5101 cttctcaaat atgcttccca gcctgctttt ctgtaacgtt caccctctac cttagcatcc
     5161 cttccctttg caaatagtcc tcttccaaca ataataatgt cagatcctgt agaaaccaca
     5221 tcatccacgg ttctatactg ttgacccaat gcatctccct tgtcatctaa acccacaccg
     5281 ggtgtcataa tcaaccaatc gtaaccttca tctcttccac ccatgtctct ttgagcaata
     5341 aagccgataa caaaatcttt gtcgctcttc gcaatgtcaa cagtaccctt agtatattct
     5401 ccagtagata gggagccctt gcatgacaat tctgctaaca tcaaaaggcc tctaggttcc
     5461 tttgttactt cttctgccgc ctgcttcaaa ccgctaacaa tacctgggcc caccacaccg
     5521 tgtgcattcg taatgtctgc ccattctgct attctgtata cacccgcaga gtactgcaat
     5581 ttgactgtat taccaatgtc agcaaatttt ctgtcctcga agagtaaaaa attgtacttg
     5641 gcggataatg cctttagcgg cttaactgtg ccctccatgg aaaaatcagt caagatatcc
     5701 acatgtgttt ttagtaaaca aattttggga cctaatgctt caactaactc cagtaattcc
     5761 ttggtggtac gaacatccaa tgaagcacac aagtttgttt gcttttcgtg catgatatta
     5821 aatagcttgg cagcaacagg actaggatga gtagcagcac gttccttata tgtagctttc
     5881 gacatgattt atcttcgttt cctgcacgtt tttgttctgt gcagttgggt taagaatact
     5941 gggcaatttc atgtttcttc aacactacat atgcgtatat ataccaatct aagtctgtgc
     6001 tccttccttc gttcttcctt ctgttcggag attaccgaat caaaaaaatt tcaaggaaac
     6061 cgtgtactgc agtgcactag tggttcggcg gccgctttgt acagaaaaaa aagaaaaatt
     6121 tgaaatataa ataacgttct taatactaac ataactatta aaaaaaataa atagggacct
     6181 agacttcagg ttgtctaact ccttcctttt cggttagagc ggatgtggga ggagggcgtg
     6241 aatgtaagcg tgacataact aattacatga tatcgacaaa ggaaaagggg cctgtttatt
     6301 tatcatcatc atctttataa tcaatatcat gatctttata atcaccatca tgatctttat
     6361 aatcagaacc tctcggcacg agtgtgtaat gtaatttgac tcctttgagc actggctcag
     6421 agtcgtcttc atcaaatttg cagcaggatc cacaagaaca acagcccttg agacaactac
     6481 agcaactggt catacagcaa agcataattg tcaccattac tatggcaatc aagccagcta
     6541 taaaacctag ccaaatgtac catggccatt ttatatactg ctcatacttt ccaagttctt
     6601 ggagatcgat gagagattca tttaaattct tggcaacctc attgaggcgg tcaatttctt
     6661 tttgaatgtt tacaactgaa gcattaatgc cagagatgtc acctaaatca acatctggtg
     6721 atgtatgatt cttaaaatat ttatctaact cctccttgaa tgagtctaat tcaggttgca
     6781 aaggatcata aactgtgttg ttgacaattc ctattacaac atcacagtta ccagacacaa
     6841 atgtgttgtc tgtagtaatg atttgtggtt cataaaaatt cctttgtgtt acaaaccagt
     6901 gtgtgccatt tgaaacaaag acaccttcac gaggaaagtg tgcttttcca tcatgacaaa
     6961 tggcaggagc agttgtgaag ttcttttctt gtgcagggac ataagtcaca tgcaagaaga
     7021 ctacaccatg aggtgctgac tgagggaagg acataagatg atagcccttt ccacaaaaat
     7081 caactctttt tgattgtcca agtacacact ctgacatttt agtagcagca agattagcag
     7141 aagctctgat ttctgcagct ctaattaatt gttgagtcac atatgtctgc aaactttgaa
     7201 gtctgcctgt gatcaaccta tcaatttgca cttcagcctc aactttgtca agacgtgaaa
     7261 ggatatcatt taaaacactt gaaattgcac caaaattgga gctaagttgt ttaacaagcg
     7321 tgtttaaagc ttgtgcattt tggttgacca catcttgaag ttttccaagt gcacttgctg
     7381 tggaagaaag tgagtcttga attttgccaa tagcactatt aaattggttg gcaatcaatt
     7441 tttggttctc atagagaaca ttctgtgtaa ctccaatacc attaaaccta taagccattt
     7501 gcatagcaaa tggtatttgt aatgcagcac ctgcaccaaa ggtccaacca gaagtgattg
     7561 tacccgctaa cagtgcagaa gtgtattgag caatcatttc atctgtgagc aaaggtggca
     7621 aaacagtaag gccgttaaac ttttgtgcac aaatgaggtc tctagcagca atatcaccaa
     7681 ggcaatcacc atattgtttg atgaagccag catctgcaag tgtcactttg ttgaaaagta
     7741 gatcttcaat aaatgacctc ttgcttggtt ttgatggatc tggtaatatt tgtgaaaaat
     7801 taaaaccacc aaaatcttta attggtggtg ttttgtaaat ttgtttgact tgtgcaaaaa
     7861 cttcttgggt gtttttgtct tgttcaacag ctattccagt taaagcacgg tttaattgtg
     7921 tacaaaaact gccatattgc aacaaaagat tgctgcattc agttgaatca ccacaaatgt
     7981 acattgtaca atctactgat gtcttggtca tagacactgg tagaatttct gtggtaacac
     8041 taatagtaaa atttgtgggt atggcaatag agttattaga gtaagcaact gaattttctg
     8101 caccaagtga catagtgtag gcaatgatgg attgactagc tacactacgt gcccgccgag
     8161 gagaattagt ctgagtctga taactagcgc atatacctgc accaatgggt atgtcacact
     8221 catatgagtt gttgacatgt tcagccccta ttaaacagcc tgcacgtgtt tgaaaaacat
     8281 tagaacctgt agaataaaca cgccaagtag gagtaagttg atctgcatga atagcaacag
     8341 ggacttctgt gcagttaaca tcctgataaa gaacagcaac ctggttagaa gtatttgttc
     8401 ctggtgttat aacactgaca ccaccaaaag aacatggtgt aatgtcaaga atctcaagtg
     8461 tctgtggatc acggacagca tcagtagtgt cagcaatgtc tctgccaaat tgttggaaag
     8521 gcagaaactt tttgttagac tcagtaagaa cacctgtgcc tgttaaacca ttgaagttga
     8581 aattgacaca tttgttttta accaaattag tagacttttt agg
//
