LOCUS       sequence-298060-       10594 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 10594)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 10594)
  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-m4046233
FEATURES             Location/Qualifiers
     source          1..10594
                     /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             6269..6286
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             6287..6304
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6349..6367)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             6718..7521
                     /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      7604..8586
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      8664..8693
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      8715..9302
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(9314..9341)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(9365..9556)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(9557..10348,10349..10417))
                     /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     10180..10199
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10418..10520)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(10521..10552)
                     /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 atggccccca ctaaggtcac attcggggat gacaccgtga tagaggttca
      481 gggttacaaa tccgttaaca tcacttttga attagatgaa agaattgaca aggtattgaa
      541 tgaaaagtgc tcagcatata ccgttgaact gggtacggag gtcaacgaat ttgcttgtgt
      601 cgttgcggat gctgttatca aaactttgca acctgtgtct gaacttctaa ccccattagg
      661 aattgacttg gatgaatggt ctatggctac ttactatcta ttcgatgaaa gtggtgagtt
      721 caagttggct tcccacatgt actgttcgtt ctacccacca gacgaagacg aagaagaagg
      781 cgactgtgaa gaagaagagt ttgaaccaag cacccaatat gaatacggta cggaagatga
      841 ttaccaaggt aagccattgg aatttggtgc cacttctgct gcattgcaac ctgaagaaga
      901 gcaagaagaa gattggttag acgatgactc tcagcaaaca gtcggtcaac aagacggttc
      961 cgaggataat caaactacca ccattcaaac aatcgttgag gtacaaccac aacttgaaat
     1021 ggaattgact ccagtcgttc agaccataga agttaactcg ttctcaggtt acttgaagct
     1081 aactgataat gtttatatta aaaacgctga catcgtcgaa gaagcaaaga aagttaagcc
     1141 taccgtcgtg gttaacgccg ctaacgttta tttgaagcat ggtggcggtg tagctggtgc
     1201 tttaaacaaa gccactaata acgctatgca agtcgaatct gacgattaca ttgctaccaa
     1261 tggtccattg aaggtgggag gctcttgcgt cttaagtgga cacaacttgg cgaaacattg
     1321 tctccacgtt gttggtccca atgttaacaa gggtgaagac attcaactgt tgaagtccgc
     1381 ctacgaaaat ttcaaccaac atgaggttct cttggcacca ttactgagtg ctggtatctt
     1441 cggcgctgac ccaattcact ctttaagggt ctgtgtggat actgtaagaa caaacgtcta
     1501 cttggccgtt ttcgataaaa acctttatga caagttagtc tcttcctttt tggaaatgaa
     1561 gtcagaaaag caagttgaac agaaaatcgc ggaaatccct aaggaagaag taaagccgtt
     1621 tataaccgaa tcgaagccct ctgttgagca aagaaagcaa gatgacaaaa agattaaagc
     1681 ttgtgttgag gaagtaacaa ctacattaga ggaaactaag tttctcacgg aaaatctatt
     1741 gttgtacata gatatcaatg gtaacttgca cccagactcc gctactttgg tcagcgatat
     1801 cgacattaca ttcttgaaga aggacgctcc atatattgtg ggtgacgttg tccaagaagg
     1861 tgttttaact gctgttgtca tcccaactaa gaaggcaggg ggtaccacgg agatgctagc
     1921 taaggccttg cgtaaagtcc cgactgataa ttatattacc acctaccctg gtcagggtct
     1981 aaacggttac acggttgaag aagctaaaac tgttctaaaa aagtgtaagt ctgcttttta
     2041 catattgcca tcaattattt ccaacgaaaa gcaagaaata ttgggtaccg tatcttggaa
     2101 tttgagagaa atgttagctc atgctgagga aactagaaag ttgatgcctg tctgcgttga
     2161 aactaaagct atcgtcagca cgatccaaag aaaatacaag ggtattaaaa tccaagaagg
     2221 cgttgtggac tatggagccc gtttctattt ctacacttcg aagaccaccg tcgcttcgtt
     2281 gatcaacacc ttgaacgatc tcaacgagac acttgttact atgccactcg gctatgtaac
     2341 tcacggtctt aacttggaag aagctgcgag atacatgcgc tctctgaaag ttcctgccac
     2401 agtctccgta tcttcaccag acgctgttac agcctataat ggatacttga cttcttcatc
     2461 aaaaacccca gaagaacatt tcatcgaaac tatctctcta gctggtagtt acaaggattg
     2521 gtcttactcc ggtcaatcca cacaactagg tatagagttt ctgaagagag gtgacaagag
     2581 tgtttactac accagcaatc caacgacctt tcatttagat ggtgaagtca ttacgttcga
     2641 taacttgaag actttgttga gtttgcggga agtccgaact atcaaggttt tcactaccgt
     2701 tgataacatt aacctccaca cccaagttgt tgatatgtct atgacttatg gtcaacagtt
     2761 cggtccgaca tatttggatg gtgctgacgt tactaaaatt aagccacaca actcccacga
     2821 agggaagaca ttctacgtgc ttccaaatga cgacacttta cgagtcgaag cctttgagta
     2881 ctaccatacc acggacccat ctttccttgg tagatacatg tccgctctga atcacactaa
     2941 aaagtggaaa tacccacaag ttaacggttt gacttctatt aagtgggccg ataataactg
     3001 ttacttggct accgccttat tgactttgca acagatagaa ctcaagttta atcctcccgc
     3061 tctacaagat gcatattata gagctagagc aggcgaagcg gctaatttct gtgctttaat
     3121 tttggcttac tgtaacaaga ccgtaggtga attgggggat gtcagagaga ctatgtccta
     3181 tcttttccag cacgcaaact tggattcatg taagagagtg ttaaatgtag tctgcaagac
     3241 ctgcggtcaa caacaaacta ccttaaaggg tgttgaagcc gttatgtaca tgggtacact
     3301 ttcttacgaa caatttaaga agggtgtcca aatcccatgt acttgtggta agcaagccac
     3361 taaatacttg gtccagcaag aaagcccttt cgttatgatg agcgccccac cagctcaata
     3421 cgagctgaag catggaactt tcacctgtgc ttctgaatat accggtaact accaatgcgg
     3481 tcactacaag catattacta gcaaagaaac tttgtactgt atcgatggtg ccttgttgac
     3541 taagtcctca gaatataagg gcccaattac tgacgtgttt tacaaagaga atagttacac
     3601 taccacaatt aagccagtaa catacaaact ggatggtgtt gtgtgtaccg agatcgaccc
     3661 taagttagat aactattaca agaaggacaa ctcttacttt accgaacaac cgatcgacct
     3721 agtcccaaat cagccatatc caaatgcttc ttttgataac ttcaagttcg tttgtgataa
     3781 cataaaattc gccgacgatt taaaccaatt gaccggttac aaaaagccgg cttcaagaga
     3841 gttgaaagtc actttcttcc cagatttaaa tggcgatgtc gttgctattg actataagca
     3901 ctacactcca tctttcaaga agggcgccaa gttgctacac aagccaattg tctggcacgt
     3961 taacaacgct accaacaagg ctacgtataa gcctaatacc tggtgtatca gatgtctatg
     4021 gtccaccaaa ccagttgaaa cgagtaactc ttttgacgtt ctgaaatcag aagatgccca
     4081 agggatggat aacttagcat gtgaagactt gaaacctgtt tcagaagaag ttgtggaaaa
     4141 tccaactatt cagaaggacg tgttggaatg caacgtgaaa acaactgaag tagttggtga
     4201 tataatcctg aagccagcta acaactcact gaagattacc gaagaagtcg gtcacaccga
     4261 cttgatggca gcctacgttg ataactcttc cctcacgatc aaaaagccaa atgaattgtc
     4321 tcgtgtactc gggctaaaga ccctagctac ccatggtcta gcggctgtca actccgtgcc
     4381 gtgggacacc atcgctaact acgctaagcc ttttttgaat aaggtcgtta gcaccacgac
     4441 aaatatagtt actagatgtt tgaacagagt gtgtaccaat tacatgcctt atttctttac
     4501 tctattgctg caattgtgca ctttcactcg ctctacaaac tccagaatta aagcatctat
     4561 gccaaccacc atcgccaaga acacggttaa gtcagtaggt aagttctgct tagaggcaag
     4621 ctttaactat ttgaagtctc caaacttctc taagttgatc aatattatta tctggttttt
     4681 acttttgagc gtttgtttgg gttccttgat ctactcaacg gctgctttgg gtgtcttgat
     4741 gtctaacttg ggtatgccga gttactgtac tggttataga gaaggttatt tgaactcaac
     4801 aaacgttact attgctacgt actgcaccgg ttccattcca tgttctgttt gtttatccgg
     4861 tttagactcg ttggacactt acccatcttt agaaaccatc caaattacca tttcctcttt
     4921 caagtgggat ttgacagctt tcggtcttgt cgctgaatgg tttttggcct atatattgtt
     4981 tactcgtttc ttctacgtct taggcttggc cgctatcatg cagttgtttt tctcttattt
     5041 tgcagtccat tttatctcaa attcgtggtt gatgtggttg attattaatc tcgttcaaat
     5101 ggctccaatt tcggcaatgg ttaggatgta cattttcttt gcttcattct attatgtatg
     5161 gaaatcttac gtccatgtgg ttgatggttg taactcctcg acctgtatga tgtgttacaa
     5221 gagaaataga gcgactaggg ttgagtgcac aactatcgtt aacggtgtca gacgttcttt
     5281 ctacgtatac gctaacggtg gtaagggctt ttgtaaactc cataactgga attgtgttaa
     5341 ttgtgatacc ttttgcgctg gttctacctt tattagtgat gaagtagcca gagacttaag
     5401 tctccaattc aaacggccaa tcaacccaac tgatcaatcc tcttatattg tcgatagtgt
     5461 caccgttaag aacggtagta tccacttata cttcgataag gccggtcaaa agacatacga
     5521 aagacactca ttgtcccact ttgttaattt agacaatttg agagctaata atactaaggg
     5581 gtctttaccc atcaacgtca tagtcttcga tggtaagtct aaatgtgaag aatcctccgc
     5641 taagtctgcg tcagtttact actctcaatt gatgtgccaa cccatcctac ttctggatca
     5701 agctttggta tctgacgttg gtgactctgc cgaggttgct gttaagatgt tcgatgctta
     5761 cgttaacaca ttttcatcaa ctttcaacgt gccaatggaa aagctgaaga ctttggtggc
     5821 taccgctgaa gcagaattgg ctaagaatgt ttctttggat aacgttttaa gtacgttcat
     5881 ttccgcagct cgtcagggtt tcgtcgactc tgacgtagaa accaaagacg tcgtcgaatg
     5941 tttgaagctc tctcatcaat ccgatattga agtcactggt gactcctgta ataactacat
     6001 gctgacgtat aacaaagtgg aaaacatgac tcctagagat ttgggagcgt gcatcgactg
     6061 ttctgctaga cacataaacg ctcaagttgc aaaatcccat aatattgctt tgatctggaa
     6121 cgttaaagat ttcatgtcac tatccgagca acttaggaaa caaatccggt ctgcggcaaa
     6181 gaaaaataac cttccattca aattaacatg tgcaacaact agacaagttg tgaacgttgt
     6241 tactactaag atcgctctga agggtggtct cgtgccgaga ggttctcacc atcatcacca
     6301 tcattaaaca ggcccctttt cctttgtcga tatcatgtaa ttagttatgt cacgcttaca
     6361 ttcacgccct cctcccacat ccgctctaac cgaaaaggaa ggagttagac aacctgaagt
     6421 ctaggtccct atttattttt tttaatagtt atgttagtat taagaacgtt atttatattt
     6481 caaatttttc ttttttttct gtacaaagcg gccgccgaac cactagtgca ctgcagtaca
     6541 cggtttcctt gaaatttttt tgattcggta atctccgaac agaaggaaga acgaaggaag
     6601 gagcacagac ttagattggt atatatacgc atatgtagtg ttgaagaaac atgaaattgc
     6661 ccagtattct taacccaact gcacagaaca aaaacgtgca ggaaacgaag ataaatcatg
     6721 tcgaaagcta catataagga acgtgctgct actcatccta gtcctgttgc tgccaagcta
     6781 tttaatatca tgcacgaaaa gcaaacaaac ttgtgtgctt cattggatgt tcgtaccacc
     6841 aaggaattac tggagttagt tgaagcatta ggtcccaaaa tttgtttact aaaaacacat
     6901 gtggatatct tgactgattt ttccatggag ggcacagtta agccgctaaa ggcattatcc
     6961 gccaagtaca attttttact cttcgaggac agaaaatttg ctgacattgg taatacagtc
     7021 aaattgcagt actctgcggg tgtatacaga atagcagaat gggcagacat tacgaatgca
     7081 cacggtgtgg tgggcccagg tattgttagc ggtttgaagc aggcggcaga agaagtaaca
     7141 aaggaaccta gaggcctttt gatgttagca gaattgtcat gcaagggctc cctatctact
     7201 ggagaatata ctaagggtac tgttgacatt gcgaagagcg acaaagattt tgttatcggc
     7261 tttattgctc aaagagacat gggtggaaga gatgaaggtt acgattggtt gattatgaca
     7321 cccggtgtgg gtttagatga caagggagat gcattgggtc aacagtatag aaccgtggat
     7381 gatgtggttt ctacaggatc tgacattatt attgttggaa gaggactatt tgcaaaggga
     7441 agggatgcta aggtagaggg tgaacgttac agaaaagcag gctgggaagc atatttgaga
     7501 agatgcggcc agcaaaacta aaaaactgta ttataagtaa atgcatgtat actaaactca
     7561 caaattagag cttcaattta attatatcag ttattacccg agtaacgaag catctgtgct
     7621 tcattttgta gaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag
     7681 ctgcattttt acagaacaga aatgcaacgc gaaagcgcta ttttaccaac gaagaatctg
     7741 tgcttcattt ttgtaaaaca aaaatgcaac gcgagagcgc taatttttca aacaaagaat
     7801 ctgagctgca tttttacaga acagaaatgc aacgcgagag cgctatttta ccaacaaaga
     7861 atctatactt cttttttgtt ctacaaaaat gcatcccgag agcgctattt ttctaacaaa
     7921 gcatcttaga ttactttttt tctcctttgt gcgctctata atgcagtctc ttgataactt
     7981 tttgcactgt aggtccgtta aggttagaag aaggctactt tggtgtctat tttctcttcc
     8041 ataaaaaaag cctgactcca cttcccgcgt ttactgatta ctagcgaagc tgcgggtgca
     8101 ttttttcaag ataaaggcat ccccgattat attctatacc gatgtggatt gcgcatactt
     8161 tgtgaacaga aagtgatagc gttgatgatt cttcattggt cagaaaatta tgaacggttt
     8221 cttctatttt gtctctatat actacgtata ggaaatgttt acattttcgt attgttttcg
     8281 attcactcta tgaatagttc ttactacaat ttttttgtct aaagagtaat actagagata
     8341 aacataaaaa atgtagaggt cgagtttaga tgcaagttca aggagcgaaa ggtggatggg
     8401 taggttatat agggatatag cacagagata tatagcaaag agatactttt gagcaatgtt
     8461 tgtggaagcg gtattcgcaa tattttagta gctcgttaca gtccggtgcg tttttggttt
     8521 tttgaaagtg cgtcatcaga gcgcttttgg ttttcaaaag cgctctgaag ttcctatact
     8581 ttctagctag agaataggaa cttcccgagc ggccgcgtgt tacaagcttg actccagcgt
     8641 aactggactg caatcaactc actggctcac cttcacgggt gggcctttct tcggtagaaa
     8701 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     8761 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     8821 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     8881 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     8941 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     9001 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     9061 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     9121 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     9181 agcgcacgag ggagcttcca ggggtaaacg cctggtatct ttatagtcct gtcgggtttc
     9241 gccacctctg acttgagcat cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     9301 aaaacgccag caacgcagaa aggcccaccc gaaggtgagc caggtgatta catttgggcc
     9361 ctcatcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat
     9421 cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga
     9481 gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt
     9541 tttcctgttt ggtcacttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     9601 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atgcgggagg
     9661 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     9721 atttatcagc aataaaccag ccagccggga gggccgagcg cagaagtgat cctgcaactt
     9781 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     9841 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     9901 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     9961 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgccaga agtaagttgg
    10021 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
    10081 ccgtgagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
    10141 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
    10201 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgtaaactc tcaaggatct
    10261 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
    10321 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
    10381 agggaataag ggcgacacgg aaatgttgaa tactcatttt agcttcctta gctcctgaaa
    10441 atctcgataa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg
    10501 aacctcttac gtgccgatca agtcaaaagc ctccggtcgg aggcttttga ctttctgcta
    10561 tggaggtcag gtatgattta aatggtcagt taga
//
