LOCUS       sequence-300412-       10641 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast expression plasmid for SARS-CoV-2 nsp3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10641)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 10641)
  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-m4046483
FEATURES             Location/Qualifiers
     source          1..10641
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     191..209
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(254..271)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(272..289)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     promoter        complement(6139..6580)
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     primer_bind     complement(6145..6168)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    6463..6580
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     terminator      6647..6678
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        6679..6781
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(6782..6850,6851..7642)
                     /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(7000..7019)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             7643..7834
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      7858..7885
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(7897..8484)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(8506..8535)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      8613..9595
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(9678..10481)
                     /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"
ORIGIN
        1 aaatttcaag gaaaccgtgt actgcagtgc actagtggtt cggcggccgc tttgtacaga
       61 aaaaaaagaa aaatttgaaa tataaataac gttcttaata ctaacataac tattaaaaaa
      121 aataaatagg gacctagact tcaggttgtc taactccttc cttttcggtt agagcggatg
      181 tgggaggagg gcgtgaatgt aagcgtgaca taactaatta catgatatcg acaaaggaaa
      241 aggggcctgt ttaatgatgg tgatgatggt gagaacctct cggcacgaga ccacccttca
      301 gagcgatctt agtagtaaca acgttcacaa cttgtctagt tgttgcacat gttaatttga
      361 atggaaggtt atttttcttt gccgcagacc ggatttgttt cctaagttgc tcggatagtg
      421 acatgaaatc tttaacgttc cagatcaaag caatattatg ggattttgca acttgagcgt
      481 ttatgtgtct agcagaacag tcgatgcacg ctcccaaatc tctaggagtc atgttttcca
      541 ctttgttata cgtcagcatg tagttattac aggagtcacc agtgacttca atatcggatt
      601 gatgagagag cttcaaacat tcgacgacgt ctttggtttc tacgtcagag tcgacgaaac
      661 cctgacgagc tgcggaaatg aacgtactta aaacgttatc caaagaaaca ttcttagcca
      721 attctgcttc agcggtagcc accaaagtct tcagcttttc cattggcacg ttgaaagttg
      781 atgaaaatgt gttaacgtaa gcatcgaaca tcttaacagc aacctcggca gagtcaccaa
      841 cgtcagatac caaagcttga tccagaagta ggatgggttg gcacatcaat tgagagtagt
      901 aaactgacgc agacttagcg gaggattctt cacatttaga cttaccatcg aagactatga
      961 cgttgatggg taaagacccc ttagtattat tagctctcaa attgtctaaa ttaacaaagt
     1021 gggacaatga gtgtctttcg tatgtctttt gaccggcctt atcgaagtat aagtggatac
     1081 taccgttctt aacggtgaca ctatcgacaa tataagagga ttgatcagtt gggttgattg
     1141 gccgtttgaa ttggagactt aagtctctgg ctacttcatc actaataaag gtagaaccag
     1201 cgcaaaaggt atcacaatta acacaattcc agttatggag tttacaaaag cccttaccac
     1261 cgttagcgta tacgtagaaa gaacgtctga caccgttaac gatagttgtg cactcaaccc
     1321 tagtcgctct atttctcttg taacacatca tacaggtcga ggagttacaa ccatcaacca
     1381 catggacgta agatttccat acataataga atgaagcaaa gaaaatgtac atcctaacca
     1441 ttgccgaaat tggagccatt tgaacgagat taataatcaa ccacatcaac cacgaatttg
     1501 agataaaatg gactgcaaaa taagagaaaa acaactgcat gatagcggcc aagcctaaga
     1561 cgtagaagaa acgagtaaac aatatatagg ccaaaaacca ttcagcgaca agaccgaaag
     1621 ctgtcaaatc ccacttgaaa gaggaaatgg taatttggat ggtttctaaa gatgggtaag
     1681 tgtccaacga gtctaaaccg gataaacaaa cagaacatgg aatggaaccg gtgcagtacg
     1741 tagcaatagt aacgtttgtt gagttcaaat aaccttctct ataaccagta cagtaactcg
     1801 gcatacccaa gttagacatc aagacaccca aagcagccgt tgagtagatc aaggaaccca
     1861 aacaaacgct caaaagtaaa aaccagataa taatattgat caacttagag aagtttggag
     1921 acttcaaata gttaaagctt gcctctaagc agaacttacc tactgactta accgtgttct
     1981 tggcgatggt ggttggcata gatgctttaa ttctggagtt tgtagagcga gtgaaagtgc
     2041 acaattgcag caatagagta aagaaataag gcatgtaatt ggtacacact ctgttcaaac
     2101 atctagtaac tatatttgtc gtggtgctaa cgaccttatt caaaaaaggc ttagcgtagt
     2161 tagcgatggt gtcccacggc acggagttga cagccgctag accatgggta gctagggtct
     2221 ttagcccgag tacacgagac aattcatttg gctttttgat cgtgagggaa gagttatcaa
     2281 cgtaggctgc catcaagtcg gtgtgaccga cttcttcggt aatcttcagt gagttgttag
     2341 ctggcttcag gattatatca ccaactactt cagttgtttt cacgttgcat tccaacacgt
     2401 ccttctgaat agttggattt tccacaactt cttctgaaac aggtttcaag tcttcacatg
     2461 ctaagttatc catcccttgg gcatcttctg atttcagaac gtcaaaagag ttactcgttt
     2521 caactggttt ggtggaccat agacatctga tacaccaggt attaggctta tacgtagcct
     2581 tgttggtagc gttgttaacg tgccagacaa ttggcttgtg tagcaacttg gcgcccttct
     2641 tgaaagatgg agtgtagtgc ttatagtcaa tagcaacgac atcgccattt aaatctggga
     2701 agaaagtgac tttcaactct cttgaagccg gctttttgta accggtcaat tggtttaaat
     2761 cgtcggcgaa ttttatgtta tcacaaacga acttgaagtt atcaaaagaa gcatttggat
     2821 atggctgatt tgggactagg tcgatcggtt gttcggtaaa gtaagagttg tccttcttgt
     2881 aatagttatc taacttaggg tcgatctcgg tacacacaac accatccagt ttgtatgtta
     2941 ctggcttaat tgtggtagtg taactattct ctttgtaaaa cacgtcagta attgggccct
     3001 tatattctga ggacttagtc aacaaggcac catcgataca gtacaaagtt tctttgctag
     3061 taatatgctt gtagtgaccg cattggtagt taccggtata ttcagaagca caggtgaaag
     3121 ttccatgctt cagctcgtat tgagctggtg gggcgctcat cataacgaaa gggctttctt
     3181 gctggaccaa gtatttagtg gcttgcttac cacaagtaca tgggatttgg acacccttct
     3241 taaattgttc gtaagaaagt gtacccatgt acataacggc ttcaacaccc tttaaggtag
     3301 tttgttgttg accgcaggtc ttgcagacta catttaacac tctcttacat gaatccaagt
     3361 ttgcgtgctg gaaaagatag gacatagtct ctctgacatc ccccaattca cctacggtct
     3421 tgttacagta agccaaaatt aaagcacaga aattagccgc ttcgcctgct ctagctctat
     3481 aatatgcatc ttgtagagcg ggaggattaa acttgagttc tatctgttgc aaagtcaata
     3541 aggcggtagc caagtaacag ttattatcgg cccacttaat agaagtcaaa ccgttaactt
     3601 gtgggtattt ccacttttta gtgtgattca gagcggacat gtatctacca aggaaagatg
     3661 ggtccgtggt atggtagtac tcaaaggctt cgactcgtaa agtgtcgtca tttggaagca
     3721 cgtagaatgt cttcccttcg tgggagttgt gtggcttaat tttagtaacg tcagcaccat
     3781 ccaaatatgt cggaccgaac tgttgaccat aagtcataga catatcaaca acttgggtgt
     3841 ggaggttaat gttatcaacg gtagtgaaaa ccttgatagt tcggacttcc cgcaaactca
     3901 acaaagtctt caagttatcg aacgtaatga cttcaccatc taaatgaaag gtcgttggat
     3961 tgctggtgta gtaaacactc ttgtcacctc tcttcagaaa ctctatacct agttgtgtgg
     4021 attgaccgga gtaagaccaa tccttgtaac taccagctag agagatagtt tcgatgaaat
     4081 gttcttctgg ggtttttgat gaagaagtca agtatccatt ataggctgta acagcgtctg
     4141 gtgaagatac ggagactgtg gcaggaactt tcagagagcg catgtatctc gcagcttctt
     4201 ccaagttaag accgtgagtt acatagccga gtggcatagt aacaagtgtc tcgttgagat
     4261 cgttcaaggt gttgatcaac gaagcgacgg tggtcttcga agtgtagaaa tagaaacggg
     4321 ctccatagtc cacaacgcct tcttggattt taataccctt gtattttctt tggatcgtgc
     4381 tgacgatagc tttagtttca acgcagacag gcatcaactt tctagtttcc tcagcatgag
     4441 ctaacatttc tctcaaattc caagatacgg tacccaatat ttcttgcttt tcgttggaaa
     4501 taattgatgg caatatgtaa aaagcagact tacacttttt tagaacagtt ttagcttctt
     4561 caaccgtgta accgtttaga ccctgaccag ggtaggtggt aatataatta tcagtcggga
     4621 ctttacgcaa ggccttagct agcatctccg tggtaccccc tgccttctta gttgggatga
     4681 caacagcagt taaaacacct tcttggacaa cgtcacccac aatatatgga gcgtccttct
     4741 tcaagaatgt aatgtcgata tcgctgacca aagtagcgga gtctgggtgc aagttaccat
     4801 tgatatctat gtacaacaat agattttccg tgagaaactt agtttcctct aatgtagttg
     4861 ttacttcctc aacacaagct ttaatctttt tgtcatcttg ctttctttgc tcaacagagg
     4921 gcttcgattc ggttataaac ggctttactt cttccttagg gatttccgcg attttctgtt
     4981 caacttgctt ttctgacttc atttccaaaa aggaagagac taacttgtca taaaggtttt
     5041 tatcgaaaac ggccaagtag acgtttgttc ttacagtatc cacacagacc cttaaagagt
     5101 gaattgggtc agcgccgaag ataccagcac tcagtaatgg tgccaagaga acctcatgtt
     5161 ggttgaaatt ttcgtaggcg gacttcaaca gttgaatgtc ttcacccttg ttaacattgg
     5221 gaccaacaac gtggagacaa tgtttcgcca agttgtgtcc acttaagacg caagagcctc
     5281 ccaccttcaa tggaccattg gtagcaatgt aatcgtcaga ttcgacttgc atagcgttat
     5341 tagtggcttt gtttaaagca ccagctacac cgccaccatg cttcaaataa acgttagcgg
     5401 cgttaaccac gacggtaggc ttaactttct ttgcttcttc gacgatgtca gcgtttttaa
     5461 tataaacatt atcagttagc ttcaagtaac ctgagaacga gttaacttct atggtctgaa
     5521 cgactggagt caattccatt tcaagttgtg gttgtacctc aacgattgtt tgaatggtgg
     5581 tagtttgatt atcctcggaa ccgtcttgtt gaccgactgt ttgctgagag tcatcgtcta
     5641 accaatcttc ttcttgctct tcttcaggtt gcaatgcagc agaagtggca ccaaattcca
     5701 atggcttacc ttggtaatca tcttccgtac cgtattcata ttgggtgctt ggttcaaact
     5761 cttcttcttc acagtcgcct tcttcttcgt cttcgtctgg tgggtagaac gaacagtaca
     5821 tgtgggaagc caacttgaac tcaccacttt catcgaatag atagtaagta gccatagacc
     5881 attcatccaa gtcaattcct aatggggtta gaagttcaga cacaggttgc aaagttttga
     5941 taacagcatc cgcaacgaca caagcaaatt cgttgacctc cgtacccagt tcaacggtat
     6001 atgctgagca cttttcattc aataccttgt caattctttc atctaattca aaagtgatgt
     6061 taacggattt gtaaccctga acctctatca cggtgtcatc cccgaatgtg accttagtgg
     6121 gggccattat agttttttct ccttgacgtt aaagtataga ggtatattaa caattttttg
     6181 ttgatacttt tatgacattt gaataagaag taatacaaac tgaaaatgtt gaaagtatta
     6241 gttaaagtgg ttatgcagct tttccattta tatatctgtt aatagatcaa aaatcatcgc
     6301 ttcgctgatt aattacccca gaaataaggc taaaaaacta atcgcattat catcctatgg
     6361 ttgttaattt gattcgttaa tttgaaggtt tgtggggcca ggttactgcc aatttttcct
     6421 cttcataacc ataaaagcta gtattgtaga atctttattg ttcggagcag tgcggcgcga
     6481 ggcacatctg cgtttcagga acgcgaccgg tgaagacgag gacgcacgga ggagagtctt
     6541 ccgtcggagg gctgtcgccc gctcggcggc ttctaatccg tacttcaata tagcaagcgg
     6601 ccgctctaac tgaccattta aatcatacct gacctccata gcagaaagtc aaaagcctcc
     6661 gaccggaggc ttttgacttg atcggcacgt aagaggttcc aactttcacc ataatgaaat
     6721 aagatcacta ccgggcgtat tttttgagtt atcgagattt tcaggagcta aggaagctaa
     6781 aatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     6841 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     6901 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gtttacgccc
     6961 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     7021 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     7081 ggttgagtac tcaccagtca cagaaaagca tctcacggat ggcatgacag taagagaatt
     7141 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tggcaacgat
     7201 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     7261 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     7321 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     7381 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggat cacttctgcg
     7441 ctcggccctc ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     7501 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgcatcg tagttatcta
     7561 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     7621 ctcactgatt aagcattggt aagtgaccaa acaggaaaaa accgccctta acatggcccg
     7681 ctttatcaga agccagacat taacgcttct ggagaaactc aacgagctgg acgcggatga
     7741 acaggcagac atctgtgaat cgcttcacga ccacgctgat gagctttacc gcagctgcct
     7801 cgcgcgtttc ggtgatgacg gtgaaaacct ctgatgaggg cccaaatgta atcacctggc
     7861 tcaccttcgg gtgggccttt ctgcgttgct ggcgtttttc cataggctcc gcccccctga
     7921 cgagcatcac aaaaatcgat gctcaagtca gaggtggcga aacccgacag gactataaag
     7981 ataccaggcg tttacccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     8041 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     8101 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     8161 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     8221 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     8281 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     8341 agtatttggt atctgcgctc tgctgaagcc agttacctcg gaaaaagagt tggtagctct
     8401 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     8461 acgcgcagaa aaaaaggatc tcaagaagat cctttgattt tctaccgaag aaaggcccac
     8521 ccgtgaaggt gagccagtga gttgattgca gtccagttac gctggagtca agcttgtaac
     8581 acgcggccgc tcgggaagtt cctattctct agctagaaag tataggaact tcagagcgct
     8641 tttgaaaacc aaaagcgctc tgatgacgca ctttcaaaaa accaaaaacg caccggactg
     8701 taacgagcta ctaaaatatt gcgaataccg cttccacaaa cattgctcaa aagtatctct
     8761 ttgctatata tctctgtgct atatccctat ataacctacc catccacctt tcgctccttg
     8821 aacttgcatc taaactcgac ctctacattt tttatgttta tctctagtat tactctttag
     8881 acaaaaaaat tgtagtaaga actattcata gagtgaatcg aaaacaatac gaaaatgtaa
     8941 acatttccta tacgtagtat atagagacaa aatagaagaa accgttcata attttctgac
     9001 caatgaagaa tcatcaacgc tatcactttc tgttcacaaa gtatgcgcaa tccacatcgg
     9061 tatagaatat aatcggggat gcctttatct tgaaaaaatg cacccgcagc ttcgctagta
     9121 atcagtaaac gcgggaagtg gagtcaggct ttttttatgg aagagaaaat agacaccaaa
     9181 gtagccttct tctaacctta acggacctac agtgcaaaaa gttatcaaga gactgcatta
     9241 tagagcgcac aaaggagaaa aaaagtaatc taagatgctt tgttagaaaa atagcgctct
     9301 cgggatgcat ttttgtagaa caaaaaagaa gtatagattc tttgttggta aaatagcgct
     9361 ctcgcgttgc atttctgttc tgtaaaaatg cagctcagat tctttgtttg aaaaattagc
     9421 gctctcgcgt tgcatttttg ttttacaaaa atgaagcaca gattcttcgt tggtaaaata
     9481 gcgctttcgc gttgcatttc tgttctgtaa aaatgcagct cagattcttt gtttgaaaaa
     9541 ttagcgctct cgcgttgcat ttttgttcta caaaatgaag cacagatgct tcgttactcg
     9601 ggtaataact gatataatta aattgaagct ctaatttgtg agtttagtat acatgcattt
     9661 acttataata cagtttttta gttttgctgg ccgcatcttc tcaaatatgc ttcccagcct
     9721 gcttttctgt aacgttcacc ctctacctta gcatcccttc cctttgcaaa tagtcctctt
     9781 ccaacaataa taatgtcaga tcctgtagaa accacatcat ccacggttct atactgttga
     9841 cccaatgcat ctcccttgtc atctaaaccc acaccgggtg tcataatcaa ccaatcgtaa
     9901 ccttcatctc ttccacccat gtctctttga gcaataaagc cgataacaaa atctttgtcg
     9961 ctcttcgcaa tgtcaacagt acccttagta tattctccag tagataggga gcccttgcat
    10021 gacaattctg ctaacatcaa aaggcctcta ggttcctttg ttacttcttc tgccgcctgc
    10081 ttcaaaccgc taacaatacc tgggcccacc acaccgtgtg cattcgtaat gtctgcccat
    10141 tctgctattc tgtatacacc cgcagagtac tgcaatttga ctgtattacc aatgtcagca
    10201 aattttctgt cctcgaagag taaaaaattg tacttggcgg ataatgcctt tagcggctta
    10261 actgtgccct ccatggaaaa atcagtcaag atatccacat gtgtttttag taaacaaatt
    10321 ttgggaccta atgcttcaac taactccagt aattccttgg tggtacgaac atccaatgaa
    10381 gcacacaagt ttgtttgctt ttcgtgcatg atattaaata gcttggcagc aacaggacta
    10441 ggatgagtag cagcacgttc cttatatgta gctttcgaca tgatttatct tcgtttcctg
    10501 cacgtttttg ttctgtgcag ttgggttaag aatactgggc aatttcatgt ttcttcaaca
    10561 ctacatatgc gtatatatac caatctaagt ctgtgctcct tccttcgttc ttccttctgt
    10621 tcggagatta ccgaatcaaa a
//
