LOCUS       sequence-296385-       10642 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 10642)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 10642)
  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-m4046390
FEATURES             Location/Qualifiers
     source          1..10642
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        15..428
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             6280..6297
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             6298..6363
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(6408..6426)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             6777..7580
                     /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      7663..8645
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      8723..8752
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      8774..9361
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(9373..9400)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(9424..9615)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(9616..10407,10408..10476))
                     /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     10239..10258
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10477..10579)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(10580..10611)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 ggtcagttag agcggccgcc ccacacacca tagcttcaaa atgtttctac tcctttttta
       61 ctcttccaga ttttctcgga ctccgcgcat cgccgtacca cttcaaaaca cccaagcaca
      121 gcatactaaa tttcccctct ttcttcctct agggtgtcgt taattacccg tactaaaggt
      181 ttggaaaaga aaaaagagac cgcctcgttt ctttttcttc gtcgaaaaag gcaataaaaa
      241 tttttatcac gtttcttttt cttgaaaatt tttttttttg atttttttct ctttcgatga
      301 cctcccattg atatttaagt taataaacgg tcttcaattt ctcaagtttc agtttcattt
      361 ttcttgttct attacaactt tttttacttc ttgctcatta gaaagaaagc atagcaatct
      421 aatctaagtt ttaattacaa aatggccccc actaaggtca cattcgggga tgacaccgtg
      481 atagaggttc agggttacaa atccgttaac atcacttttg aattagatga aagaattgac
      541 aaggtattga atgaaaagtg ctcagcatat accgttgaac tgggtacgga ggtcaacgaa
      601 tttgcttgtg tcgttgcgga tgctgttatc aaaactttgc aacctgtgtc tgaacttcta
      661 accccattag gaattgactt ggatgaatgg tctatggcta cttactatct attcgatgaa
      721 agtggtgagt tcaagttggc ttcccacatg tactgttcgt tctacccacc agacgaagac
      781 gaagaagaag gcgactgtga agaagaagag tttgaaccaa gcacccaata tgaatacggt
      841 acggaagatg attaccaagg taagccattg gaatttggtg ccacttctgc tgcattgcaa
      901 cctgaagaag agcaagaaga agattggtta gacgatgact ctcagcaaac agtcggtcaa
      961 caagacggtt ccgaggataa tcaaactacc accattcaaa caatcgttga ggtacaacca
     1021 caacttgaaa tggaattgac tccagtcgtt cagaccatag aagttaactc gttctcaggt
     1081 tacttgaagc taactgataa tgtttatatt aaaaacgctg acatcgtcga agaagcaaag
     1141 aaagttaagc ctaccgtcgt ggttaacgcc gctaacgttt atttgaagca tggtggcggt
     1201 gtagctggtg ctttaaacaa agccactaat aacgctatgc aagtcgaatc tgacgattac
     1261 attgctacca atggtccatt gaaggtggga ggctcttgcg tcttaagtgg acacaacttg
     1321 gcgaaacatt gtctccacgt tgttggtccc aatgttaaca agggtgaaga cattcaactg
     1381 ttgaagtccg cctacgaaaa tttcaaccaa catgaggttc tcttggcacc attactgagt
     1441 gctggtatct tcggcgctga cccaattcac tctttaaggg tctgtgtgga tactgtaaga
     1501 acaaacgtct acttggccgt tttcgataaa aacctttatg acaagttagt ctcttccttt
     1561 ttggaaatga agtcagaaaa gcaagttgaa cagaaaatcg cggaaatccc taaggaagaa
     1621 gtaaagccgt ttataaccga atcgaagccc tctgttgagc aaagaaagca agatgacaaa
     1681 aagattaaag cttgtgttga ggaagtaaca actacattag aggaaactaa gtttctcacg
     1741 gaaaatctat tgttgtacat agatatcaat ggtaacttgc acccagactc cgctactttg
     1801 gtcagcgata tcgacattac attcttgaag aaggacgctc catatattgt gggtgacgtt
     1861 gtccaagaag gtgttttaac tgctgttgtc atcccaacta agaaggcagg gggtaccacg
     1921 gagatgctag ctaaggcctt gcgtaaagtc ccgactgata attatattac cacctaccct
     1981 ggtcagggtc taaacggtta cacggttgaa gaagctaaaa ctgttctaaa aaagtgtaag
     2041 tctgcttttt acatattgcc atcaattatt tccaacgaaa agcaagaaat attgggtacc
     2101 gtatcttgga atttgagaga aatgttagct catgctgagg aaactagaaa gttgatgcct
     2161 gtctgcgttg aaactaaagc tatcgtcagc acgatccaaa gaaaatacaa gggtattaaa
     2221 atccaagaag gcgttgtgga ctatggagcc cgtttctatt tctacacttc gaagaccacc
     2281 gtcgcttcgt tgatcaacac cttgaacgat ctcaacgaga cacttgttac tatgccactc
     2341 ggctatgtaa ctcacggtct taacttggaa gaagctgcga gatacatgcg ctctctgaaa
     2401 gttcctgcca cagtctccgt atcttcacca gacgctgtta cagcctataa tggatacttg
     2461 acttcttcat caaaaacccc agaagaacat ttcatcgaaa ctatctctct agctggtagt
     2521 tacaaggatt ggtcttactc cggtcaatcc acacaactag gtatagagtt tctgaagaga
     2581 ggtgacaaga gtgtttacta caccagcaat ccaacgacct ttcatttaga tggtgaagtc
     2641 attacgttcg ataacttgaa gactttgttg agtttgcggg aagtccgaac tatcaaggtt
     2701 ttcactaccg ttgataacat taacctccac acccaagttg ttgatatgtc tatgacttat
     2761 ggtcaacagt tcggtccgac atatttggat ggtgctgacg ttactaaaat taagccacac
     2821 aactcccacg aagggaagac attctacgtg cttccaaatg acgacacttt acgagtcgaa
     2881 gcctttgagt actaccatac cacggaccca tctttccttg gtagatacat gtccgctctg
     2941 aatcacacta aaaagtggaa atacccacaa gttaacggtt tgacttctat taagtgggcc
     3001 gataataact gttacttggc taccgcctta ttgactttgc aacagataga actcaagttt
     3061 aatcctcccg ctctacaaga tgcatattat agagctagag caggcgaagc ggctaatttc
     3121 tgtgctttaa ttttggctta ctgtaacaag accgtaggtg aattggggga tgtcagagag
     3181 actatgtcct atcttttcca gcacgcaaac ttggattcat gtaagagagt gttaaatgta
     3241 gtctgcaaga cctgcggtca acaacaaact accttaaagg gtgttgaagc cgttatgtac
     3301 atgggtacac tttcttacga acaatttaag aagggtgtcc aaatcccatg tacttgtggt
     3361 aagcaagcca ctaaatactt ggtccagcaa gaaagccctt tcgttatgat gagcgcccca
     3421 ccagctcaat acgagctgaa gcatggaact ttcacctgtg cttctgaata taccggtaac
     3481 taccaatgcg gtcactacaa gcatattact agcaaagaaa ctttgtactg tatcgatggt
     3541 gccttgttga ctaagtcctc agaatataag ggcccaatta ctgacgtgtt ttacaaagag
     3601 aatagttaca ctaccacaat taagccagta acatacaaac tggatggtgt tgtgtgtacc
     3661 gagatcgacc ctaagttaga taactattac aagaaggaca actcttactt taccgaacaa
     3721 ccgatcgacc tagtcccaaa tcagccatat ccaaatgctt cttttgataa cttcaagttc
     3781 gtttgtgata acataaaatt cgccgacgat ttaaaccaat tgaccggtta caaaaagccg
     3841 gcttcaagag agttgaaagt cactttcttc ccagatttaa atggcgatgt cgttgctatt
     3901 gactataagc actacactcc atctttcaag aagggcgcca agttgctaca caagccaatt
     3961 gtctggcacg ttaacaacgc taccaacaag gctacgtata agcctaatac ctggtgtatc
     4021 agatgtctat ggtccaccaa accagttgaa acgagtaact cttttgacgt tctgaaatca
     4081 gaagatgccc aagggatgga taacttagca tgtgaagact tgaaacctgt ttcagaagaa
     4141 gttgtggaaa atccaactat tcagaaggac gtgttggaat gcaacgtgaa aacaactgaa
     4201 gtagttggtg atataatcct gaagccagct aacaactcac tgaagattac cgaagaagtc
     4261 ggtcacaccg acttgatggc agcctacgtt gataactctt ccctcacgat caaaaagcca
     4321 aatgaattgt ctcgtgtact cgggctaaag accctagcta cccatggtct agcggctgtc
     4381 aactccgtgc cgtgggacac catcgctaac tacgctaagc cttttttgaa taaggtcgtt
     4441 agcaccacga caaatatagt tactagatgt ttgaacagag tgtgtaccaa ttacatgcct
     4501 tatttcttta ctctattgct gcaattgtgc actttcactc gctctacaaa ctccagaatt
     4561 aaagcatcta tgccaaccac catcgccaag aacacggtta agtcagtagg taagttctgc
     4621 ttagaggcaa gctttaacta tttgaagtct ccaaacttct ctaagttgat caatattatt
     4681 atctggtttt tacttttgag cgtttgtttg ggttccttga tctactcaac ggctgctttg
     4741 ggtgtcttga tgtctaactt gggtatgccg agttactgta ctggttatag agaaggttat
     4801 ttgaactcaa caaacgttac tattgctacg tactgcaccg gttccattcc atgttctgtt
     4861 tgtttatccg gtttagactc gttggacact tacccatctt tagaaaccat ccaaattacc
     4921 atttcctctt tcaagtggga tttgacagct ttcggtcttg tcgctgaatg gtttttggcc
     4981 tatatattgt ttactcgttt cttctacgtc ttaggcttgg ccgctatcat gcagttgttt
     5041 ttctcttatt ttgcagtcca ttttatctca aattcgtggt tgatgtggtt gattattaat
     5101 ctcgttcaaa tggctccaat ttcggcaatg gttaggatgt acattttctt tgcttcattc
     5161 tattatgtat ggaaatctta cgtccatgtg gttgatggtt gtaactcctc gacctgtatg
     5221 atgtgttaca agagaaatag agcgactagg gttgagtgca caactatcgt taacggtgtc
     5281 agacgttctt tctacgtata cgctaacggt ggtaagggct tttgtaaact ccataactgg
     5341 aattgtgtta attgtgatac cttttgcgct ggttctacct ttattagtga tgaagtagcc
     5401 agagacttaa gtctccaatt caaacggcca atcaacccaa ctgatcaatc ctcttatatt
     5461 gtcgatagtg tcaccgttaa gaacggtagt atccacttat acttcgataa ggccggtcaa
     5521 aagacatacg aaagacactc attgtcccac tttgttaatt tagacaattt gagagctaat
     5581 aatactaagg ggtctttacc catcaacgtc atagtcttcg atggtaagtc taaatgtgaa
     5641 gaatcctccg ctaagtctgc gtcagtttac tactctcaat tgatgtgcca acccatccta
     5701 cttctggatc aagctttggt atctgacgtt ggtgactctg ccgaggttgc tgttaagatg
     5761 ttcgatgctt acgttaacac attttcatca actttcaacg tgccaatgga aaagctgaag
     5821 actttggtgg ctaccgctga agcagaattg gctaagaatg tttctttgga taacgtttta
     5881 agtacgttca tttccgcagc tcgtcagggt ttcgtcgact ctgacgtaga aaccaaagac
     5941 gtcgtcgaat gtttgaagct ctctcatcaa tccgatattg aagtcactgg tgactcctgt
     6001 aataactaca tgctgacgta taacaaagtg gaaaacatga ctcctagaga tttgggagcg
     6061 tgcatcgact gttctgctag acacataaac gctcaagttg caaaatccca taatattgct
     6121 ttgatctgga acgttaaaga tttcatgtca ctatccgagc aacttaggaa acaaatccgg
     6181 tctgcggcaa agaaaaataa ccttccattc aaattaacat gtgcaacaac tagacaagtt
     6241 gtgaacgttg ttactactaa gatcgctctg aagggtggtc tcgtgccgag aggttctgat
     6301 tataaagatc atgatggtga ttataaagat catgatattg attataaaga tgatgatgat
     6361 aaataaacag gccccttttc ctttgtcgat atcatgtaat tagttatgtc acgcttacat
     6421 tcacgccctc ctcccacatc cgctctaacc gaaaaggaag gagttagaca acctgaagtc
     6481 taggtcccta tttatttttt ttaatagtta tgttagtatt aagaacgtta tttatatttc
     6541 aaatttttct tttttttctg tacaaagcgg ccgccgaacc actagtgcac tgcagtacac
     6601 ggtttccttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg
     6661 agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc
     6721 cagtattctt aacccaactg cacagaacaa aaacgtgcag gaaacgaaga taaatcatgt
     6781 cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct gccaagctat
     6841 ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt cgtaccacca
     6901 aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta aaaacacatg
     6961 tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag gcattatccg
     7021 ccaagtacaa ttttttactc ttcgaggaca gaaaatttgc tgacattggt aatacagtca
     7081 aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt acgaatgcac
     7141 acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa gaagtaacaa
     7201 aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc ctatctactg
     7261 gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt gttatcggct
     7321 ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg attatgacac
     7381 ccggtgtggg tttagatgac aagggagatg cattgggtca acagtataga accgtggatg
     7441 atgtggtttc tacaggatct gacattatta ttgttggaag aggactattt gcaaagggaa
     7501 gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca tatttgagaa
     7561 gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata ctaaactcac
     7621 aaattagagc ttcaatttaa ttatatcagt tattacccga gtaacgaagc atctgtgctt
     7681 cattttgtag aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc
     7741 tgcattttta cagaacagaa atgcaacgcg aaagcgctat tttaccaacg aagaatctgt
     7801 gcttcatttt tgtaaaacaa aaatgcaacg cgagagcgct aatttttcaa acaaagaatc
     7861 tgagctgcat ttttacagaa cagaaatgca acgcgagagc gctattttac caacaaagaa
     7921 tctatacttc ttttttgttc tacaaaaatg catcccgaga gcgctatttt tctaacaaag
     7981 catcttagat tacttttttt ctcctttgtg cgctctataa tgcagtctct tgataacttt
     8041 ttgcactgta ggtccgttaa ggttagaaga aggctacttt ggtgtctatt ttctcttcca
     8101 taaaaaaagc ctgactccac ttcccgcgtt tactgattac tagcgaagct gcgggtgcat
     8161 tttttcaaga taaaggcatc cccgattata ttctataccg atgtggattg cgcatacttt
     8221 gtgaacagaa agtgatagcg ttgatgattc ttcattggtc agaaaattat gaacggtttc
     8281 ttctattttg tctctatata ctacgtatag gaaatgttta cattttcgta ttgttttcga
     8341 ttcactctat gaatagttct tactacaatt tttttgtcta aagagtaata ctagagataa
     8401 acataaaaaa tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag gtggatgggt
     8461 aggttatata gggatatagc acagagatat atagcaaaga gatacttttg agcaatgttt
     8521 gtggaagcgg tattcgcaat attttagtag ctcgttacag tccggtgcgt ttttggtttt
     8581 ttgaaagtgc gtcatcagag cgcttttggt tttcaaaagc gctctgaagt tcctatactt
     8641 tctagctaga gaataggaac ttcccgagcg gccgcgtgtt acaagcttga ctccagcgta
     8701 actggactgc aatcaactca ctggctcacc ttcacgggtg ggcctttctt cggtagaaaa
     8761 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     8821 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     8881 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     8941 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     9001 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     9061 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     9121 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     9181 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     9241 gcgcacgagg gagcttccag gggtaaacgc ctggtatctt tatagtcctg tcgggtttcg
     9301 ccacctctga cttgagcatc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     9361 aaacgccagc aacgcagaaa ggcccacccg aaggtgagcc aggtgattac atttgggccc
     9421 tcatcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc
     9481 agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag
     9541 tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt
     9601 ttcctgtttg gtcacttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     9661 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tgcgggaggg
     9721 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     9781 tttatcagca ataaaccagc cagccgggag ggccgagcgc agaagtgatc ctgcaacttt
     9841 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     9901 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     9961 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
    10021 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgccagaa gtaagttggc
    10081 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
    10141 cgtgagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
    10201 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
    10261 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgtaaactct caaggatctt
    10321 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
    10381 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
    10441 gggaataagg gcgacacgga aatgttgaat actcatttta gcttccttag ctcctgaaaa
    10501 tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt gaaagttgga
    10561 acctcttacg tgccgatcaa gtcaaaagcc tccggtcgga ggcttttgac tttctgctat
    10621 ggaggtcagg tatgatttaa at
//
