LOCUS       sequence-300493-       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"
     terminator      992..1021
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      1043..1630
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(1642..1669)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1693..1884)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(1885..2676,2677..2745))
                     /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     2508..2527
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2746..2848)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(2849..2880)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        2926..3339
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     complement(9235..9253)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             9604..10407
                     /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      join(10490..10558,1..914)
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 aatctgagct gcatttttac agaacagaaa tgcaacgcga aagcgctatt ttaccaacga
       61 agaatctgtg cttcattttt gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa
      121 caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgagagcg ctattttacc
      181 aacaaagaat ctatacttct tttttgttct acaaaaatgc atcccgagag cgctattttt
      241 ctaacaaagc atcttagatt actttttttc tcctttgtgc gctctataat gcagtctctt
      301 gataactttt tgcactgtag gtccgttaag gttagaagaa ggctactttg gtgtctattt
      361 tctcttccat aaaaaaagcc tgactccact tcccgcgttt actgattact agcgaagctg
      421 cgggtgcatt ttttcaagat aaaggcatcc ccgattatat tctataccga tgtggattgc
      481 gcatactttg tgaacagaaa gtgatagcgt tgatgattct tcattggtca gaaaattatg
      541 aacggtttct tctattttgt ctctatatac tacgtatagg aaatgtttac attttcgtat
      601 tgttttcgat tcactctatg aatagttctt actacaattt ttttgtctaa agagtaatac
      661 tagagataaa cataaaaaat gtagaggtcg agtttagatg caagttcaag gagcgaaagg
      721 tggatgggta ggttatatag ggatatagca cagagatata tagcaaagag atacttttga
      781 gcaatgtttg tggaagcggt attcgcaata ttttagtagc tcgttacagt ccggtgcgtt
      841 tttggttttt tgaaagtgcg tcatcagagc gcttttggtt ttcaaaagcg ctctgaagtt
      901 cctatacttt ctagctagag aataggaact tcccgagcgg ccgcgtgtta caagcttgac
      961 tccagcgtaa ctggactgca atcaactcac tggctcacct tcacgggtgg gcctttcttc
     1021 ggtagaaaat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     1081 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     1141 tttttccgag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     1201 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     1261 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     1321 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     1381 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     1441 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     1501 aacaggagag cgcacgaggg agcttccagg ggtaaacgcc tggtatcttt atagtcctgt
     1561 cgggtttcgc cacctctgac ttgagcatcg atttttgtga tgctcgtcag gggggcggag
     1621 cctatggaaa aacgccagca acgcagaaag gcccacccga aggtgagcca ggtgattaca
     1681 tttgggccct catcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta
     1741 aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat ccgcgtccag
     1801 ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag
     1861 ggcggttttt tcctgtttgg tcacttacca atgcttaatc agtgaggcac ctatctcagc
     1921 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     1981 gcgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     2041 ggctccagat ttatcagcaa taaaccagcc agccgggagg gccgagcgca gaagtgatcc
     2101 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     2161 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     2221 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     2281 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgccagaag
     2341 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     2401 catgccatcc gtgagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     2461 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     2521 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gtaaactctc
     2581 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     2641 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     2701 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcattttag cttccttagc
     2761 tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg
     2821 aaagttggaa cctcttacgt gccgatcaag tcaaaagcct ccggtcggag gcttttgact
     2881 ttctgctatg gaggtcaggt atgatttaaa tggtcagtta gagcggccgc cccacacacc
     2941 atagcttcaa aatgtttcta ctcctttttt actcttccag attttctcgg actccgcgca
     3001 tcgccgtacc acttcaaaac acccaagcac agcatactaa atttcccctc tttcttcctc
     3061 tagggtgtcg ttaattaccc gtactaaagg tttggaaaag aaaaaagaga ccgcctcgtt
     3121 tctttttctt cgtcgaaaaa ggcaataaaa atttttatca cgtttctttt tcttgaaaat
     3181 tttttttttt gatttttttc tctttcgatg acctcccatt gatatttaag ttaataaacg
     3241 gtcttcaatt tctcaagttt cagtttcatt tttcttgttc tattacaact ttttttactt
     3301 cttgctcatt agaaagaaag catagcaatc taatctaagt tttaattaca aaatggcccc
     3361 cactaaggtc acattcgggg atgacaccgt gatagaggtt cagggttaca aatccgttaa
     3421 catcactttt gaattagatg aaagaattga caaggtattg aatgaaaagt gctcagcata
     3481 taccgttgaa ctgggtacgg aggtcaacga atttgcttgt gtcgttgcgg atgctgttat
     3541 caaaactttg caacctgtgt ctgaacttct aaccccatta ggaattgact tggatgaatg
     3601 gtctatggct acttactatc tattcgatga aagtggtgag ttcaagttgg cttcccacat
     3661 gtactgttcg ttctacccac cagacgaaga cgaagaagaa ggcgactgtg aagaagaaga
     3721 gtttgaacca agcacccaat atgaatacgg tacggaagat gattaccaag gtaagccatt
     3781 ggaatttggt gccacttctg ctgcattgca acctgaagaa gagcaagaag aagattggtt
     3841 agacgatgac tctcagcaaa cagtcggtca acaagacggt tccgaggata atcaaactac
     3901 caccattcaa acaatcgttg aggtacaacc acaacttgaa atggaattga ctccagtcgt
     3961 tcagaccata gaagttaact cgttctcagg ttacttgaag ctaactgata atgtttatat
     4021 taaaaacgct gacatcgtcg aagaagcaaa gaaagttaag cctaccgtcg tggttaacgc
     4081 cgctaacgtt tatttgaagc atggtggcgg tgtagctggt gctttaaaca aagccactaa
     4141 taacgctatg caagtcgaat ctgacgatta cattgctacc aatggtccat tgaaggtggg
     4201 aggctcttgc gtcttaagtg gacacaactt ggcgaaacat tgtctccacg ttgttggtcc
     4261 caatgttaac aagggtgaag acattcaact gttgaagtcc gcctacgaaa atttcaacca
     4321 acatgaggtt ctcttggcac cattactgag tgctggtatc ttcggcgctg acccaattca
     4381 ctctttaagg gtctgtgtgg atactgtaag aacaaacgtc tacttggccg ttttcgataa
     4441 aaacctttat gacaagttag tctcttcctt tttggaaatg aagtcagaaa agcaagttga
     4501 acagaaaatc gcggaaatcc ctaaggaaga agtaaagccg tttataaccg aatcgaagcc
     4561 ctctgttgag caaagaaagc aagatgacaa aaagattaaa gcttgtgttg aggaagtaac
     4621 aactacatta gaggaaacta agtttctcac ggaaaatcta ttgttgtaca tagatatcaa
     4681 tggtaacttg cacccagact ccgctacttt ggtcagcgat atcgacatta cattcttgaa
     4741 gaaggacgct ccatatattg tgggtgacgt tgtccaagaa ggtgttttaa ctgctgttgt
     4801 catcccaact aagaaggcag ggggtaccac ggagatgcta gctaaggcct tgcgtaaagt
     4861 cccgactgat aattatatta ccacctaccc tggtcagggt ctaaacggtt acacggttga
     4921 agaagctaaa actgttctaa aaaagtgtaa gtctgctttt tacatattgc catcaattat
     4981 ttccaacgaa aagcaagaaa tattgggtac cgtatcttgg aatttgagag aaatgttagc
     5041 tcatgctgag gaaactagaa agttgatgcc tgtctgcgtt gaaactaaag ctatcgtcag
     5101 cacgatccaa agaaaataca agggtattaa aatccaagaa ggcgttgtgg actatggagc
     5161 ccgtttctat ttctacactt cgaagaccac cgtcgcttcg ttgatcaaca ccttgaacga
     5221 tctcaacgag acacttgtta ctatgccact cggctatgta actcacggtc ttaacttgga
     5281 agaagctgcg agatacatgc gctctctgaa agttcctgcc acagtctccg tatcttcacc
     5341 agacgctgtt acagcctata atggatactt gacttcttca tcaaaaaccc cagaagaaca
     5401 tttcatcgaa actatctctc tagctggtag ttacaaggat tggtcttact ccggtcaatc
     5461 cacacaacta ggtatagagt ttctgaagag aggtgacaag agtgtttact acaccagcaa
     5521 tccaacgacc tttcatttag atggtgaagt cattacgttc gataacttga agactttgtt
     5581 gagtttgcgg gaagtccgaa ctatcaaggt tttcactacc gttgataaca ttaacctcca
     5641 cacccaagtt gttgatatgt ctatgactta tggtcaacag ttcggtccga catatttgga
     5701 tggtgctgac gttactaaaa ttaagccaca caactcccac gaagggaaga cattctacgt
     5761 gcttccaaat gacgacactt tacgagtcga agcctttgag tactaccata ccacggaccc
     5821 atctttcctt ggtagataca tgtccgctct gaatcacact aaaaagtgga aatacccaca
     5881 agttaacggt ttgacttcta ttaagtgggc cgataataac tgttacttgg ctaccgcctt
     5941 attgactttg caacagatag aactcaagtt taatcctccc gctctacaag atgcatatta
     6001 tagagctaga gcaggcgaag cggctaattt ctgtgcttta attttggctt actgtaacaa
     6061 gaccgtaggt gaattggggg atgtcagaga gactatgtcc tatcttttcc agcacgcaaa
     6121 cttggattca tgtaagagag tgttaaatgt agtctgcaag acctgcggtc aacaacaaac
     6181 taccttaaag ggtgttgaag ccgttatgta catgggtaca ctttcttacg aacaatttaa
     6241 gaagggtgtc caaatcccat gtacttgtgg taagcaagcc actaaatact tggtccagca
     6301 agaaagccct ttcgttatga tgagcgcccc accagctcaa tacgagctga agcatggaac
     6361 tttcacctgt gcttctgaat ataccggtaa ctaccaatgc ggtcactaca agcatattac
     6421 tagcaaagaa actttgtact gtatcgatgg tgccttgttg actaagtcct cagaatataa
     6481 gggcccaatt actgacgtgt tttacaaaga gaatagttac actaccacaa ttaagccagt
     6541 aacatacaaa ctggatggtg ttgtgtgtac cgagatcgac cctaagttag ataactatta
     6601 caagaaggac aactcttact ttaccgaaca accgatcgac ctagtcccaa atcagccata
     6661 tccaaatgct tcttttgata acttcaagtt cgtttgtgat aacataaaat tcgccgacga
     6721 tttaaaccaa ttgaccggtt acaaaaagcc ggcttcaaga gagttgaaag tcactttctt
     6781 cccagattta aatggcgatg tcgttgctat tgactataag cactacactc catctttcaa
     6841 gaagggcgcc aagttgctac acaagccaat tgtctggcac gttaacaacg ctaccaacaa
     6901 ggctacgtat aagcctaata cctggtgtat cagatgtcta tggtccacca aaccagttga
     6961 aacgagtaac tcttttgacg ttctgaaatc agaagatgcc caagggatgg ataacttagc
     7021 atgtgaagac ttgaaacctg tttcagaaga agttgtggaa aatccaacta ttcagaagga
     7081 cgtgttggaa tgcaacgtga aaacaactga agtagttggt gatataatcc tgaagccagc
     7141 taacaactca ctgaagatta ccgaagaagt cggtcacacc gacttgatgg cagcctacgt
     7201 tgataactct tccctcacga tcaaaaagcc aaatgaattg tctcgtgtac tcgggctaaa
     7261 gaccctagct acccatggtc tagcggctgt caactccgtg ccgtgggaca ccatcgctaa
     7321 ctacgctaag ccttttttga ataaggtcgt tagcaccacg acaaatatag ttactagatg
     7381 tttgaacaga gtgtgtacca attacatgcc ttatttcttt actctattgc tgcaattgtg
     7441 cactttcact cgctctacaa actccagaat taaagcatct atgccaacca ccatcgccaa
     7501 gaacacggtt aagtcagtag gtaagttctg cttagaggca agctttaact atttgaagtc
     7561 tccaaacttc tctaagttga tcaatattat tatctggttt ttacttttga gcgtttgttt
     7621 gggttccttg atctactcaa cggctgcttt gggtgtcttg atgtctaact tgggtatgcc
     7681 gagttactgt actggttata gagaaggtta tttgaactca acaaacgtta ctattgctac
     7741 gtactgcacc ggttccattc catgttctgt ttgtttatcc ggtttagact cgttggacac
     7801 ttacccatct ttagaaacca tccaaattac catttcctct ttcaagtggg atttgacagc
     7861 tttcggtctt gtcgctgaat ggtttttggc ctatatattg tttactcgtt tcttctacgt
     7921 cttaggcttg gccgctatca tgcagttgtt tttctcttat tttgcagtcc attttatctc
     7981 aaattcgtgg ttgatgtggt tgattattaa tctcgttcaa atggctccaa tttcggcaat
     8041 ggttaggatg tacattttct ttgcttcatt ctattatgta tggaaatctt acgtccatgt
     8101 ggttgatggt tgtaactcct cgacctgtat gatgtgttac aagagaaata gagcgactag
     8161 ggttgagtgc acaactatcg ttaacggtgt cagacgttct ttctacgtat acgctaacgg
     8221 tggtaagggc ttttgtaaac tccataactg gaattgtgtt aattgtgata ccttttgcgc
     8281 tggttctacc tttattagtg atgaagtagc cagagactta agtctccaat tcaaacggcc
     8341 aatcaaccca actgatcaat cctcttatat tgtcgatagt gtcaccgtta agaacggtag
     8401 tatccactta tacttcgata aggccggtca aaagacatac gaaagacact cattgtccca
     8461 ctttgttaat ttagacaatt tgagagctaa taatactaag gggtctttac ccatcaacgt
     8521 catagtcttc gatggtaagt ctaaatgtga agaatcctcc gctaagtctg cgtcagttta
     8581 ctactctcaa ttgatgtgcc aacccatcct acttctggat caagctttgg tatctgacgt
     8641 tggtgactct gccgaggttg ctgttaagat gttcgatgct tacgttaaca cattttcatc
     8701 aactttcaac gtgccaatgg aaaagctgaa gactttggtg gctaccgctg aagcagaatt
     8761 ggctaagaat gtttctttgg ataacgtttt aagtacgttc atttccgcag ctcgtcaggg
     8821 tttcgtcgac tctgacgtag aaaccaaaga cgtcgtcgaa tgtttgaagc tctctcatca
     8881 atccgatatt gaagtcactg gtgactcctg taataactac atgctgacgt ataacaaagt
     8941 ggaaaacatg actcctagag atttgggagc gtgcatcgac tgttctgcta gacacataaa
     9001 cgctcaagtt gcaaaatccc ataatattgc tttgatctgg aacgttaaag atttcatgtc
     9061 actatccgag caacttagga aacaaatccg gtctgcggca aagaaaaata accttccatt
     9121 caaattaaca tgtgcaacaa ctagacaagt tgtgaacgtt gttactacta agatcgctct
     9181 gaagggtggt taaacaggcc ccttttcctt tgtcgatatc atgtaattag ttatgtcacg
     9241 cttacattca cgccctcctc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc
     9301 tgaagtctag gtccctattt atttttttta atagttatgt tagtattaag aacgttattt
     9361 atatttcaaa tttttctttt ttttctgtac aaagcggccg ccgaaccact agtgcactgc
     9421 agtacacggt ttccttgaaa tttttttgat tcggtaatct ccgaacagaa ggaagaacga
     9481 aggaaggagc acagacttag attggtatat atacgcatat gtagtgttga agaaacatga
     9541 aattgcccag tattcttaac ccaactgcac agaacaaaaa cgtgcaggaa acgaagataa
     9601 atcatgtcga aagctacata taaggaacgt gctgctactc atcctagtcc tgttgctgcc
     9661 aagctattta atatcatgca cgaaaagcaa acaaacttgt gtgcttcatt ggatgttcgt
     9721 accaccaagg aattactgga gttagttgaa gcattaggtc ccaaaatttg tttactaaaa
     9781 acacatgtgg atatcttgac tgatttttcc atggagggca cagttaagcc gctaaaggca
     9841 ttatccgcca agtacaattt tttactcttc gaggacagaa aatttgctga cattggtaat
     9901 acagtcaaat tgcagtactc tgcgggtgta tacagaatag cagaatgggc agacattacg
     9961 aatgcacacg gtgtggtggg cccaggtatt gttagcggtt tgaagcaggc ggcagaagaa
    10021 gtaacaaagg aacctagagg ccttttgatg ttagcagaat tgtcatgcaa gggctcccta
    10081 tctactggag aatatactaa gggtactgtt gacattgcga agagcgacaa agattttgtt
    10141 atcggcttta ttgctcaaag agacatgggt ggaagagatg aaggttacga ttggttgatt
    10201 atgacacccg gtgtgggttt agatgacaag ggagatgcat tgggtcaaca gtatagaacc
    10261 gtggatgatg tggtttctac aggatctgac attattattg ttggaagagg actatttgca
    10321 aagggaaggg atgctaaggt agagggtgaa cgttacagaa aagcaggctg ggaagcatat
    10381 ttgagaagat gcggccagca aaactaaaaa actgtattat aagtaaatgc atgtatacta
    10441 aactcacaaa ttagagcttc aatttaatta tatcagttat tacccgagta acgaagcatc
    10501 tgtgcttcat tttgtagaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaag
//
