LOCUS       sequence-297618-       10558 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 10558)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 10558)
  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-m4046632
FEATURES             Location/Qualifiers
     source          1..10558
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     complement(6313..6331)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             6682..7485
                     /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      7568..8550
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      8628..8657
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      8679..9266
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(9278..9305)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(9329..9520)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(9521..10312,10313..10381))
                     /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     10144..10163
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10382..10484)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(10485..10516)
                     /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 agggtggtta aacaggcccc ttttcctttg tcgatatcat
     6301 gtaattagtt atgtcacgct tacattcacg ccctcctccc acatccgctc taaccgaaaa
     6361 ggaaggagtt agacaacctg aagtctaggt ccctatttat tttttttaat agttatgtta
     6421 gtattaagaa cgttatttat atttcaaatt tttctttttt ttctgtacaa agcggccgcc
     6481 gaaccactag tgcactgcag tacacggttt ccttgaaatt tttttgattc ggtaatctcc
     6541 gaacagaagg aagaacgaag gaaggagcac agacttagat tggtatatat acgcatatgt
     6601 agtgttgaag aaacatgaaa ttgcccagta ttcttaaccc aactgcacag aacaaaaacg
     6661 tgcaggaaac gaagataaat catgtcgaaa gctacatata aggaacgtgc tgctactcat
     6721 cctagtcctg ttgctgccaa gctatttaat atcatgcacg aaaagcaaac aaacttgtgt
     6781 gcttcattgg atgttcgtac caccaaggaa ttactggagt tagttgaagc attaggtccc
     6841 aaaatttgtt tactaaaaac acatgtggat atcttgactg atttttccat ggagggcaca
     6901 gttaagccgc taaaggcatt atccgccaag tacaattttt tactcttcga ggacagaaaa
     6961 tttgctgaca ttggtaatac agtcaaattg cagtactctg cgggtgtata cagaatagca
     7021 gaatgggcag acattacgaa tgcacacggt gtggtgggcc caggtattgt tagcggtttg
     7081 aagcaggcgg cagaagaagt aacaaaggaa cctagaggcc ttttgatgtt agcagaattg
     7141 tcatgcaagg gctccctatc tactggagaa tatactaagg gtactgttga cattgcgaag
     7201 agcgacaaag attttgttat cggctttatt gctcaaagag acatgggtgg aagagatgaa
     7261 ggttacgatt ggttgattat gacacccggt gtgggtttag atgacaaggg agatgcattg
     7321 ggtcaacagt atagaaccgt ggatgatgtg gtttctacag gatctgacat tattattgtt
     7381 ggaagaggac tatttgcaaa gggaagggat gctaaggtag agggtgaacg ttacagaaaa
     7441 gcaggctggg aagcatattt gagaagatgc ggccagcaaa actaaaaaac tgtattataa
     7501 gtaaatgcat gtatactaaa ctcacaaatt agagcttcaa tttaattata tcagttatta
     7561 cccgagtaac gaagcatctg tgcttcattt tgtagaacaa aaatgcaacg cgagagcgct
     7621 aatttttcaa acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgaaagc
     7681 gctattttac caacgaagaa tctgtgcttc atttttgtaa aacaaaaatg caacgcgaga
     7741 gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg
     7801 agagcgctat tttaccaaca aagaatctat acttcttttt tgttctacaa aaatgcatcc
     7861 cgagagcgct atttttctaa caaagcatct tagattactt tttttctcct ttgtgcgctc
     7921 tataatgcag tctcttgata actttttgca ctgtaggtcc gttaaggtta gaagaaggct
     7981 actttggtgt ctattttctc ttccataaaa aaagcctgac tccacttccc gcgtttactg
     8041 attactagcg aagctgcggg tgcatttttt caagataaag gcatccccga ttatattcta
     8101 taccgatgtg gattgcgcat actttgtgaa cagaaagtga tagcgttgat gattcttcat
     8161 tggtcagaaa attatgaacg gtttcttcta ttttgtctct atatactacg tataggaaat
     8221 gtttacattt tcgtattgtt ttcgattcac tctatgaata gttcttacta caattttttt
     8281 gtctaaagag taatactaga gataaacata aaaaatgtag aggtcgagtt tagatgcaag
     8341 ttcaaggagc gaaaggtgga tgggtaggtt atatagggat atagcacaga gatatatagc
     8401 aaagagatac ttttgagcaa tgtttgtgga agcggtattc gcaatatttt agtagctcgt
     8461 tacagtccgg tgcgtttttg gttttttgaa agtgcgtcat cagagcgctt ttggttttca
     8521 aaagcgctct gaagttccta tactttctag ctagagaata ggaacttccc gagcggccgc
     8581 gtgttacaag cttgactcca gcgtaactgg actgcaatca actcactggc tcaccttcac
     8641 gggtgggcct ttcttcggta gaaaatcaaa ggatcttctt gagatccttt ttttctgcgc
     8701 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     8761 caagagctac caactctttt tccgaggtaa ctggcttcag cagagcgcag ataccaaata
     8821 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     8881 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     8941 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     9001 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     9061 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     9121 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggta aacgcctggt
     9181 atctttatag tcctgtcggg tttcgccacc tctgacttga gcatcgattt ttgtgatgct
     9241 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc agaaaggccc acccgaaggt
     9301 gagccaggtg attacatttg ggccctcatc agaggttttc accgtcatca ccgaaacgcg
     9361 cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct
     9421 gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa
     9481 agcgggccat gttaagggcg gttttttcct gtttggtcac ttaccaatgc ttaatcagtg
     9541 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     9601 tgtagataac tacgatgcgg gagggcttac catctggccc cagtgctgca atgataccgc
     9661 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc gggagggccg
     9721 agcgcagaag tgatcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     9781 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     9841 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     9901 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     9961 cgatcgttgc cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
    10021 ataattctct tactgtcatg ccatccgtga gatgcttttc tgtgactggt gagtactcaa
    10081 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
    10141 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
    10201 cggggcgtaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
    10261 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
    10321 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
    10381 ttttagcttc cttagctcct gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc
    10441 ttatttcatt atggtgaaag ttggaacctc ttacgtgccg atcaagtcaa aagcctccgg
    10501 tcggaggctt ttgactttct gctatggagg tcaggtatga tttaaatggt cagttaga
//
