LOCUS       sequence-298244-       10600 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 10600)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 10600)
  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-m4046555
FEATURES             Location/Qualifiers
     source          1..10600
                     /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..6310
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(6355..6373)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             6724..7527
                     /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      7610..8592
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      8670..8699
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      8721..9308
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(9320..9347)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(9371..9562)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(9563..10354,10355..10423))
                     /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     10186..10205
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10424..10526)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(10527..10558)
                     /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 ggttcttgga gccatccgca
     6301 gtttgaaaaa taaacaggcc ccttttcctt tgtcgatatc atgtaattag ttatgtcacg
     6361 cttacattca cgccctcctc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc
     6421 tgaagtctag gtccctattt atttttttta atagttatgt tagtattaag aacgttattt
     6481 atatttcaaa tttttctttt ttttctgtac aaagcggccg ccgaaccact agtgcactgc
     6541 agtacacggt ttccttgaaa tttttttgat tcggtaatct ccgaacagaa ggaagaacga
     6601 aggaaggagc acagacttag attggtatat atacgcatat gtagtgttga agaaacatga
     6661 aattgcccag tattcttaac ccaactgcac agaacaaaaa cgtgcaggaa acgaagataa
     6721 atcatgtcga aagctacata taaggaacgt gctgctactc atcctagtcc tgttgctgcc
     6781 aagctattta atatcatgca cgaaaagcaa acaaacttgt gtgcttcatt ggatgttcgt
     6841 accaccaagg aattactgga gttagttgaa gcattaggtc ccaaaatttg tttactaaaa
     6901 acacatgtgg atatcttgac tgatttttcc atggagggca cagttaagcc gctaaaggca
     6961 ttatccgcca agtacaattt tttactcttc gaggacagaa aatttgctga cattggtaat
     7021 acagtcaaat tgcagtactc tgcgggtgta tacagaatag cagaatgggc agacattacg
     7081 aatgcacacg gtgtggtggg cccaggtatt gttagcggtt tgaagcaggc ggcagaagaa
     7141 gtaacaaagg aacctagagg ccttttgatg ttagcagaat tgtcatgcaa gggctcccta
     7201 tctactggag aatatactaa gggtactgtt gacattgcga agagcgacaa agattttgtt
     7261 atcggcttta ttgctcaaag agacatgggt ggaagagatg aaggttacga ttggttgatt
     7321 atgacacccg gtgtgggttt agatgacaag ggagatgcat tgggtcaaca gtatagaacc
     7381 gtggatgatg tggtttctac aggatctgac attattattg ttggaagagg actatttgca
     7441 aagggaaggg atgctaaggt agagggtgaa cgttacagaa aagcaggctg ggaagcatat
     7501 ttgagaagat gcggccagca aaactaaaaa actgtattat aagtaaatgc atgtatacta
     7561 aactcacaaa ttagagcttc aatttaatta tatcagttat tacccgagta acgaagcatc
     7621 tgtgcttcat tttgtagaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa
     7681 tctgagctgc atttttacag aacagaaatg caacgcgaaa gcgctatttt accaacgaag
     7741 aatctgtgct tcatttttgt aaaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca
     7801 aagaatctga gctgcatttt tacagaacag aaatgcaacg cgagagcgct attttaccaa
     7861 caaagaatct atacttcttt tttgttctac aaaaatgcat cccgagagcg ctatttttct
     7921 aacaaagcat cttagattac tttttttctc ctttgtgcgc tctataatgc agtctcttga
     7981 taactttttg cactgtaggt ccgttaaggt tagaagaagg ctactttggt gtctattttc
     8041 tcttccataa aaaaagcctg actccacttc ccgcgtttac tgattactag cgaagctgcg
     8101 ggtgcatttt ttcaagataa aggcatcccc gattatattc tataccgatg tggattgcgc
     8161 atactttgtg aacagaaagt gatagcgttg atgattcttc attggtcaga aaattatgaa
     8221 cggtttcttc tattttgtct ctatatacta cgtataggaa atgtttacat tttcgtattg
     8281 ttttcgattc actctatgaa tagttcttac tacaattttt ttgtctaaag agtaatacta
     8341 gagataaaca taaaaaatgt agaggtcgag tttagatgca agttcaagga gcgaaaggtg
     8401 gatgggtagg ttatataggg atatagcaca gagatatata gcaaagagat acttttgagc
     8461 aatgtttgtg gaagcggtat tcgcaatatt ttagtagctc gttacagtcc ggtgcgtttt
     8521 tggttttttg aaagtgcgtc atcagagcgc ttttggtttt caaaagcgct ctgaagttcc
     8581 tatactttct agctagagaa taggaacttc ccgagcggcc gcgtgttaca agcttgactc
     8641 cagcgtaact ggactgcaat caactcactg gctcaccttc acgggtgggc ctttcttcgg
     8701 tagaaaatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     8761 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     8821 tttccgaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     8881 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     8941 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     9001 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     9061 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     9121 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     9181 caggagagcg cacgagggag cttccagggg taaacgcctg gtatctttat agtcctgtcg
     9241 ggtttcgcca cctctgactt gagcatcgat ttttgtgatg ctcgtcaggg gggcggagcc
     9301 tatggaaaaa cgccagcaac gcagaaaggc ccacccgaag gtgagccagg tgattacatt
     9361 tgggccctca tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa
     9421 gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct
     9481 cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg
     9541 cggttttttc ctgtttggtc acttaccaat gcttaatcag tgaggcacct atctcagcga
     9601 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatgc
     9661 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     9721 ctccagattt atcagcaata aaccagccag ccgggagggc cgagcgcaga agtgatcctg
     9781 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     9841 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     9901 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     9961 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gccagaagta
    10021 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
    10081 tgccatccgt gagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
    10141 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
    10201 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcgt aaactctcaa
    10261 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
    10321 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
    10381 caaaaaaggg aataagggcg acacggaaat gttgaatact cattttagct tccttagctc
    10441 ctgaaaatct cgataactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa
    10501 agttggaacc tcttacgtgc cgatcaagtc aaaagcctcc ggtcggaggc ttttgacttt
    10561 ctgctatgga ggtcaggtat gatttaaatg gtcagttaga
//
