LOCUS       sequence-300317-       10605 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 10605)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 10605)
  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-m4046663
FEATURES             Location/Qualifiers
     source          1..10605
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        25..466
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    25..142
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     437..460
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     complement(6360..6378)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             6729..7532
                     /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      7615..8597
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      8675..8704
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      8726..9313
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(9325..9352)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(9376..9567)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(9568..10359,10360..10428))
                     /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     10191..10210
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10429..10531)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(10532..10563)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgctt gctatattga agtacggatt agaagccgcc gagcgggcga cagccctccg
       61 acggaagact ctcctccgtg cgtcctcgtc ttcaccggtc gcgttcctga aacgcagatg
      121 tgcctcgcgc cgcactgctc cgaacaataa agattctaca atactagctt ttatggttat
      181 gaagaggaaa aattggcagt aacctggccc cacaaacctt caaattaacg aatcaaatta
      241 acaaccatag gatgataatg cgattagttt tttagcctta tttctggggt aattaatcag
      301 cgaagcgatg atttttgatc tattaacaga tatataaatg gaaaagctgc ataaccactt
      361 taactaatac tttcaacatt ttcagtttgt attacttctt attcaaatgt cataaaagta
      421 tcaacaaaaa attgttaata tacctctata ctttaacgtc aaggagaaaa aactataatg
      481 gcccccacta aggtcacatt cggggatgac accgtgatag aggttcaggg ttacaaatcc
      541 gttaacatca cttttgaatt agatgaaaga attgacaagg tattgaatga aaagtgctca
      601 gcatataccg ttgaactggg tacggaggtc aacgaatttg cttgtgtcgt tgcggatgct
      661 gttatcaaaa ctttgcaacc tgtgtctgaa cttctaaccc cattaggaat tgacttggat
      721 gaatggtcta tggctactta ctatctattc gatgaaagtg gtgagttcaa gttggcttcc
      781 cacatgtact gttcgttcta cccaccagac gaagacgaag aagaaggcga ctgtgaagaa
      841 gaagagtttg aaccaagcac ccaatatgaa tacggtacgg aagatgatta ccaaggtaag
      901 ccattggaat ttggtgccac ttctgctgca ttgcaacctg aagaagagca agaagaagat
      961 tggttagacg atgactctca gcaaacagtc ggtcaacaag acggttccga ggataatcaa
     1021 actaccacca ttcaaacaat cgttgaggta caaccacaac ttgaaatgga attgactcca
     1081 gtcgttcaga ccatagaagt taactcgttc tcaggttact tgaagctaac tgataatgtt
     1141 tatattaaaa acgctgacat cgtcgaagaa gcaaagaaag ttaagcctac cgtcgtggtt
     1201 aacgccgcta acgtttattt gaagcatggt ggcggtgtag ctggtgcttt aaacaaagcc
     1261 actaataacg ctatgcaagt cgaatctgac gattacattg ctaccaatgg tccattgaag
     1321 gtgggaggct cttgcgtctt aagtggacac aacttggcga aacattgtct ccacgttgtt
     1381 ggtcccaatg ttaacaaggg tgaagacatt caactgttga agtccgccta cgaaaatttc
     1441 aaccaacatg aggttctctt ggcaccatta ctgagtgctg gtatcttcgg cgctgaccca
     1501 attcactctt taagggtctg tgtggatact gtaagaacaa acgtctactt ggccgttttc
     1561 gataaaaacc tttatgacaa gttagtctct tcctttttgg aaatgaagtc agaaaagcaa
     1621 gttgaacaga aaatcgcgga aatccctaag gaagaagtaa agccgtttat aaccgaatcg
     1681 aagccctctg ttgagcaaag aaagcaagat gacaaaaaga ttaaagcttg tgttgaggaa
     1741 gtaacaacta cattagagga aactaagttt ctcacggaaa atctattgtt gtacatagat
     1801 atcaatggta acttgcaccc agactccgct actttggtca gcgatatcga cattacattc
     1861 ttgaagaagg acgctccata tattgtgggt gacgttgtcc aagaaggtgt tttaactgct
     1921 gttgtcatcc caactaagaa ggcagggggt accacggaga tgctagctaa ggccttgcgt
     1981 aaagtcccga ctgataatta tattaccacc taccctggtc agggtctaaa cggttacacg
     2041 gttgaagaag ctaaaactgt tctaaaaaag tgtaagtctg ctttttacat attgccatca
     2101 attatttcca acgaaaagca agaaatattg ggtaccgtat cttggaattt gagagaaatg
     2161 ttagctcatg ctgaggaaac tagaaagttg atgcctgtct gcgttgaaac taaagctatc
     2221 gtcagcacga tccaaagaaa atacaagggt attaaaatcc aagaaggcgt tgtggactat
     2281 ggagcccgtt tctatttcta cacttcgaag accaccgtcg cttcgttgat caacaccttg
     2341 aacgatctca acgagacact tgttactatg ccactcggct atgtaactca cggtcttaac
     2401 ttggaagaag ctgcgagata catgcgctct ctgaaagttc ctgccacagt ctccgtatct
     2461 tcaccagacg ctgttacagc ctataatgga tacttgactt cttcatcaaa aaccccagaa
     2521 gaacatttca tcgaaactat ctctctagct ggtagttaca aggattggtc ttactccggt
     2581 caatccacac aactaggtat agagtttctg aagagaggtg acaagagtgt ttactacacc
     2641 agcaatccaa cgacctttca tttagatggt gaagtcatta cgttcgataa cttgaagact
     2701 ttgttgagtt tgcgggaagt ccgaactatc aaggttttca ctaccgttga taacattaac
     2761 ctccacaccc aagttgttga tatgtctatg acttatggtc aacagttcgg tccgacatat
     2821 ttggatggtg ctgacgttac taaaattaag ccacacaact cccacgaagg gaagacattc
     2881 tacgtgcttc caaatgacga cactttacga gtcgaagcct ttgagtacta ccataccacg
     2941 gacccatctt tccttggtag atacatgtcc gctctgaatc acactaaaaa gtggaaatac
     3001 ccacaagtta acggtttgac ttctattaag tgggccgata ataactgtta cttggctacc
     3061 gccttattga ctttgcaaca gatagaactc aagtttaatc ctcccgctct acaagatgca
     3121 tattatagag ctagagcagg cgaagcggct aatttctgtg ctttaatttt ggcttactgt
     3181 aacaagaccg taggtgaatt gggggatgtc agagagacta tgtcctatct tttccagcac
     3241 gcaaacttgg attcatgtaa gagagtgtta aatgtagtct gcaagacctg cggtcaacaa
     3301 caaactacct taaagggtgt tgaagccgtt atgtacatgg gtacactttc ttacgaacaa
     3361 tttaagaagg gtgtccaaat cccatgtact tgtggtaagc aagccactaa atacttggtc
     3421 cagcaagaaa gccctttcgt tatgatgagc gccccaccag ctcaatacga gctgaagcat
     3481 ggaactttca cctgtgcttc tgaatatacc ggtaactacc aatgcggtca ctacaagcat
     3541 attactagca aagaaacttt gtactgtatc gatggtgcct tgttgactaa gtcctcagaa
     3601 tataagggcc caattactga cgtgttttac aaagagaata gttacactac cacaattaag
     3661 ccagtaacat acaaactgga tggtgttgtg tgtaccgaga tcgaccctaa gttagataac
     3721 tattacaaga aggacaactc ttactttacc gaacaaccga tcgacctagt cccaaatcag
     3781 ccatatccaa atgcttcttt tgataacttc aagttcgttt gtgataacat aaaattcgcc
     3841 gacgatttaa accaattgac cggttacaaa aagccggctt caagagagtt gaaagtcact
     3901 ttcttcccag atttaaatgg cgatgtcgtt gctattgact ataagcacta cactccatct
     3961 ttcaagaagg gcgccaagtt gctacacaag ccaattgtct ggcacgttaa caacgctacc
     4021 aacaaggcta cgtataagcc taatacctgg tgtatcagat gtctatggtc caccaaacca
     4081 gttgaaacga gtaactcttt tgacgttctg aaatcagaag atgcccaagg gatggataac
     4141 ttagcatgtg aagacttgaa acctgtttca gaagaagttg tggaaaatcc aactattcag
     4201 aaggacgtgt tggaatgcaa cgtgaaaaca actgaagtag ttggtgatat aatcctgaag
     4261 ccagctaaca actcactgaa gattaccgaa gaagtcggtc acaccgactt gatggcagcc
     4321 tacgttgata actcttccct cacgatcaaa aagccaaatg aattgtctcg tgtactcggg
     4381 ctaaagaccc tagctaccca tggtctagcg gctgtcaact ccgtgccgtg ggacaccatc
     4441 gctaactacg ctaagccttt tttgaataag gtcgttagca ccacgacaaa tatagttact
     4501 agatgtttga acagagtgtg taccaattac atgccttatt tctttactct attgctgcaa
     4561 ttgtgcactt tcactcgctc tacaaactcc agaattaaag catctatgcc aaccaccatc
     4621 gccaagaaca cggttaagtc agtaggtaag ttctgcttag aggcaagctt taactatttg
     4681 aagtctccaa acttctctaa gttgatcaat attattatct ggtttttact tttgagcgtt
     4741 tgtttgggtt ccttgatcta ctcaacggct gctttgggtg tcttgatgtc taacttgggt
     4801 atgccgagtt actgtactgg ttatagagaa ggttatttga actcaacaaa cgttactatt
     4861 gctacgtact gcaccggttc cattccatgt tctgtttgtt tatccggttt agactcgttg
     4921 gacacttacc catctttaga aaccatccaa attaccattt cctctttcaa gtgggatttg
     4981 acagctttcg gtcttgtcgc tgaatggttt ttggcctata tattgtttac tcgtttcttc
     5041 tacgtcttag gcttggccgc tatcatgcag ttgtttttct cttattttgc agtccatttt
     5101 atctcaaatt cgtggttgat gtggttgatt attaatctcg ttcaaatggc tccaatttcg
     5161 gcaatggtta ggatgtacat tttctttgct tcattctatt atgtatggaa atcttacgtc
     5221 catgtggttg atggttgtaa ctcctcgacc tgtatgatgt gttacaagag aaatagagcg
     5281 actagggttg agtgcacaac tatcgttaac ggtgtcagac gttctttcta cgtatacgct
     5341 aacggtggta agggcttttg taaactccat aactggaatt gtgttaattg tgataccttt
     5401 tgcgctggtt ctacctttat tagtgatgaa gtagccagag acttaagtct ccaattcaaa
     5461 cggccaatca acccaactga tcaatcctct tatattgtcg atagtgtcac cgttaagaac
     5521 ggtagtatcc acttatactt cgataaggcc ggtcaaaaga catacgaaag acactcattg
     5581 tcccactttg ttaatttaga caatttgaga gctaataata ctaaggggtc tttacccatc
     5641 aacgtcatag tcttcgatgg taagtctaaa tgtgaagaat cctccgctaa gtctgcgtca
     5701 gtttactact ctcaattgat gtgccaaccc atcctacttc tggatcaagc tttggtatct
     5761 gacgttggtg actctgccga ggttgctgtt aagatgttcg atgcttacgt taacacattt
     5821 tcatcaactt tcaacgtgcc aatggaaaag ctgaagactt tggtggctac cgctgaagca
     5881 gaattggcta agaatgtttc tttggataac gttttaagta cgttcatttc cgcagctcgt
     5941 cagggtttcg tcgactctga cgtagaaacc aaagacgtcg tcgaatgttt gaagctctct
     6001 catcaatccg atattgaagt cactggtgac tcctgtaata actacatgct gacgtataac
     6061 aaagtggaaa acatgactcc tagagatttg ggagcgtgca tcgactgttc tgctagacac
     6121 ataaacgctc aagttgcaaa atcccataat attgctttga tctggaacgt taaagatttc
     6181 atgtcactat ccgagcaact taggaaacaa atccggtctg cggcaaagaa aaataacctt
     6241 ccattcaaat taacatgtgc aacaactaga caagttgtga acgttgttac tactaagatc
     6301 gctctgaagg gtggttaaac aggccccttt tcctttgtcg atatcatgta attagttatg
     6361 tcacgcttac attcacgccc tcctcccaca tccgctctaa ccgaaaagga aggagttaga
     6421 caacctgaag tctaggtccc tatttatttt ttttaatagt tatgttagta ttaagaacgt
     6481 tatttatatt tcaaattttt cttttttttc tgtacaaagc ggccgccgaa ccactagtgc
     6541 actgcagtac acggtttcct tgaaattttt ttgattcggt aatctccgaa cagaaggaag
     6601 aacgaaggaa ggagcacaga cttagattgg tatatatacg catatgtagt gttgaagaaa
     6661 catgaaattg cccagtattc ttaacccaac tgcacagaac aaaaacgtgc aggaaacgaa
     6721 gataaatcat gtcgaaagct acatataagg aacgtgctgc tactcatcct agtcctgttg
     6781 ctgccaagct atttaatatc atgcacgaaa agcaaacaaa cttgtgtgct tcattggatg
     6841 ttcgtaccac caaggaatta ctggagttag ttgaagcatt aggtcccaaa atttgtttac
     6901 taaaaacaca tgtggatatc ttgactgatt tttccatgga gggcacagtt aagccgctaa
     6961 aggcattatc cgccaagtac aattttttac tcttcgagga cagaaaattt gctgacattg
     7021 gtaatacagt caaattgcag tactctgcgg gtgtatacag aatagcagaa tgggcagaca
     7081 ttacgaatgc acacggtgtg gtgggcccag gtattgttag cggtttgaag caggcggcag
     7141 aagaagtaac aaaggaacct agaggccttt tgatgttagc agaattgtca tgcaagggct
     7201 ccctatctac tggagaatat actaagggta ctgttgacat tgcgaagagc gacaaagatt
     7261 ttgttatcgg ctttattgct caaagagaca tgggtggaag agatgaaggt tacgattggt
     7321 tgattatgac acccggtgtg ggtttagatg acaagggaga tgcattgggt caacagtata
     7381 gaaccgtgga tgatgtggtt tctacaggat ctgacattat tattgttgga agaggactat
     7441 ttgcaaaggg aagggatgct aaggtagagg gtgaacgtta cagaaaagca ggctgggaag
     7501 catatttgag aagatgcggc cagcaaaact aaaaaactgt attataagta aatgcatgta
     7561 tactaaactc acaaattaga gcttcaattt aattatatca gttattaccc gagtaacgaa
     7621 gcatctgtgc ttcattttgt agaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca
     7681 aagaatctga gctgcatttt tacagaacag aaatgcaacg cgaaagcgct attttaccaa
     7741 cgaagaatct gtgcttcatt tttgtaaaac aaaaatgcaa cgcgagagcg ctaatttttc
     7801 aaacaaagaa tctgagctgc atttttacag aacagaaatg caacgcgaga gcgctatttt
     7861 accaacaaag aatctatact tcttttttgt tctacaaaaa tgcatcccga gagcgctatt
     7921 tttctaacaa agcatcttag attacttttt ttctcctttg tgcgctctat aatgcagtct
     7981 cttgataact ttttgcactg taggtccgtt aaggttagaa gaaggctact ttggtgtcta
     8041 ttttctcttc cataaaaaaa gcctgactcc acttcccgcg tttactgatt actagcgaag
     8101 ctgcgggtgc attttttcaa gataaaggca tccccgatta tattctatac cgatgtggat
     8161 tgcgcatact ttgtgaacag aaagtgatag cgttgatgat tcttcattgg tcagaaaatt
     8221 atgaacggtt tcttctattt tgtctctata tactacgtat aggaaatgtt tacattttcg
     8281 tattgttttc gattcactct atgaatagtt cttactacaa tttttttgtc taaagagtaa
     8341 tactagagat aaacataaaa aatgtagagg tcgagtttag atgcaagttc aaggagcgaa
     8401 aggtggatgg gtaggttata tagggatata gcacagagat atatagcaaa gagatacttt
     8461 tgagcaatgt ttgtggaagc ggtattcgca atattttagt agctcgttac agtccggtgc
     8521 gtttttggtt ttttgaaagt gcgtcatcag agcgcttttg gttttcaaaa gcgctctgaa
     8581 gttcctatac tttctagcta gagaatagga acttcccgag cggccgcgtg ttacaagctt
     8641 gactccagcg taactggact gcaatcaact cactggctca ccttcacggg tgggcctttc
     8701 ttcggtagaa aatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     8761 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     8821 ctctttttcc gaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     8881 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     8941 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     9001 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     9061 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     9121 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     9181 cggaacagga gagcgcacga gggagcttcc aggggtaaac gcctggtatc tttatagtcc
     9241 tgtcgggttt cgccacctct gacttgagca tcgatttttg tgatgctcgt caggggggcg
     9301 gagcctatgg aaaaacgcca gcaacgcaga aaggcccacc cgaaggtgag ccaggtgatt
     9361 acatttgggc cctcatcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg
     9421 gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc
     9481 cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt
     9541 aagggcggtt ttttcctgtt tggtcactta ccaatgctta atcagtgagg cacctatctc
     9601 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     9661 gatgcgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     9721 accggctcca gatttatcag caataaacca gccagccggg agggccgagc gcagaagtga
     9781 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     9841 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     9901 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     9961 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgccag
    10021 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
    10081 tgtcatgcca tccgtgagat gcttttctgt gactggtgag tactcaacca agtcattctg
    10141 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
    10201 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgtaaact
    10261 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
    10321 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
    10381 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcattt tagcttcctt
    10441 agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta tttcattatg
    10501 gtgaaagttg gaacctctta cgtgccgatc aagtcaaaag cctccggtcg gaggcttttg
    10561 actttctgct atggaggtca ggtatgattt aaatggtcag ttaga
//
