LOCUS       sequence-297963-        7602 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 RNA-dependent RNA 
            polymerase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7602)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 7602)
  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-m4046581
FEATURES             Location/Qualifiers
     source          1..7602
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        25..466
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    25..142
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     437..460
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             3277..3294
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             3295..3312
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3357..3375)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             3726..4529
                     /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      4612..5594
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      5672..5701
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      5723..6310
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(6322..6349)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(6373..6564)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6565..7356,7357..7425))
                     /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     7188..7207
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7426..7528)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(7529..7560)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgctt gctatattga agtacggatt agaagccgcc gagcgggcga cagccctccg
       61 acggaagact ctcctccgtg cgtcctcgtc ttcaccggtc gcgttcctga aacgcagatg
      121 tgcctcgcgc cgcactgctc cgaacaataa agattctaca atactagctt ttatggttat
      181 gaagaggaaa aattggcagt aacctggccc cacaaacctt caaattaacg aatcaaatta
      241 acaaccatag gatgataatg cgattagttt tttagcctta tttctggggt aattaatcag
      301 cgaagcgatg atttttgatc tattaacaga tatataaatg gaaaagctgc ataaccactt
      361 taactaatac tttcaacatt ttcagtttgt attacttctt attcaaatgt cataaaagta
      421 tcaacaaaaa attgttaata tacctctata ctttaacgtc aaggagaaaa aactataatg
      481 tccgccgatg cacaatcttt cttaaataga gtctgcggcg tgagtgctgc tcgtctaacc
      541 ccttgtggta cgggtacttc aacagacgta gtttacaggg cgtttgatat ttataacgac
      601 aaggtcgctg gtttcgctaa atttttgaag accaactgtt gtagattcca ggagaaggat
      661 gaagacgaca atcttatcga ttcgtacttc gttgttaaaa gacatacttt ttccaactat
      721 caacacgaag aaacaatata caatttgctc aaggactgtc cagccgtcgc taaacacgat
      781 tttttcaagt tcagaattga tggggacatg gtgccacata tcagcagaca acggttgacc
      841 aagtacacta tggctgattt agtttacgcc ttgcgacact ttgacgaagg aaactgtgat
      901 actctgaagg aaattttggt tacctataac tgctgtgacg atgattactt caacaaaaag
      961 gactggtacg acttcgtaga gaatcccgac atcttgaggg tttacgctaa cttaggtgaa
     1021 agagtcagac aagctttgtt aaagacagtc caattttgcg atgctatgag aaatgcgggt
     1081 attgttggtg ttttgacttt ggataatcaa gatttgaacg gtaactggta tgacttcggt
     1141 gattttatcc agactacccc gggttctggg gtaccagttg tcgattctta ctattctctg
     1201 ttaatgccaa tactcaccct aactagagca ttgactgccg aatcccatgt tgacactgat
     1261 ttaactaagc catacattaa atgggacctt ctaaagtacg atttcaccga agaacgttta
     1321 aagttgttcg acagatattt taaatactgg gaccaaacct accaccctaa ttgtgtcaac
     1381 tgtttggacg acagatgtat tcttcactgc gctaacttta atgtgttgtt ctcgactgtt
     1441 tttccaccaa cctctttcgg tccgctggtt cgaaagattt ttgttgacgg tgttccattc
     1501 gtggtcagta cgggatatca tttcagagaa ttaggggtcg ttcataacca agacgtgaac
     1561 ttgcactcat caagattgtc cttcaaggaa ttgttggtct acgcagctga tcctgcaatg
     1621 cacgccgctt ctggaaactt attgctggat aagagaacaa cctgtttcag cgttgccgct
     1681 cttactaata acgtcgcgtt ccaaactgta aagccgggta actttaataa agacttctac
     1741 gattttgctg ttagcaaggg ttttttcaaa gaaggttcta gtgttgagtt gaaacacttt
     1801 ttctttgctc aagacggcaa tgctgctatc agtgattatg actattaccg ttacaactta
     1861 ccaacaatgt gtgatatcag acagttgctt ttcgtcgtcg aagttgttga taaatatttt
     1921 gactgttatg atggtggttg catcaatgca aatcaagtca ttgtcaacaa cttagataag
     1981 tctgctggct tcccattcaa caagtggggt aaagccagat tgtactatga ttccatgtcc
     2041 tacgaggacc aagatgcttt gttcgcgtac accaagagaa acgttattcc tacaattact
     2101 caaatgaatc taaagtacgc tatctctgca aaaaatcggg ccagaaccgt tgctggtgtt
     2161 tctatctgtt ctacgatgac taaccgtcaa tttcatcaga agctattgaa atccatagct
     2221 gctactagag gtgctactgt agtcatcggt acctcaaagt tctatggtgg ttggcacaat
     2281 atgctcaaga ccgtttactc tgatgtcgaa aacccacatc ttatgggttg ggactaccct
     2341 aagtgtgata gagccatgcc aaatatgcta agaattatgg catccttggt cttggctaga
     2401 aagcatacca catgctgctc attgtcccac cgtttttaca gattggccaa cgaatgtgct
     2461 caagtccttt ctgaaatggt tatgtgtggt ggtagtctat atgtaaagcc aggcggaact
     2521 tcgtctggag atgccaccac tgcttacgct aactcggttt ttaacatttg tcaagccgtt
     2581 accgccaacg taaacgcttt gctgtctacg gatggtaaca agatcgctga taagtatgtg
     2641 cgcaacttgc aacatagact ttatgaatgt ttgtatagaa atcgagatgt cgatactgac
     2701 ttcgttaacg aattttacgc gtacttaaga aagcacttct ccatgatgat tttgtcagac
     2761 gacgctgttg tttgtttcaa ttctacttac gctagccaag gtttggtcgc ttcaatcaaa
     2821 aacttcaagt ctgttttata ctaccaaaac aatgttttta tgtccgaggc aaaatgctgg
     2881 acggaaacag atttgactaa gggtccacat gaattttgta gtcaacacac aatgttagtg
     2941 aaacaagggg acgactacgt ttacctaccc tacccagatc caagcagaat tttgggtgcc
     3001 ggttgtttcg tggacgacat agtcaagacc gacggtacct tgatgatcga acggtttgtc
     3061 tctctggcca tagacgctta cccattgact aagcacccta accaagaata tgctgatgtt
     3121 tttcatttat atcttcaata cattagaaaa ttacatgacg aattgaccgg tcacatgtta
     3181 gatatgtact ctgtgatgtt gacgaacgat aatacgtcac gttactggga acccgaattt
     3241 tacgaagcga tgtacactcc acacacagta ttgcagctcg tgccgagagg ttctcaccat
     3301 catcaccatc attaaacagg ccccttttcc tttgtcgata tcatgtaatt agttatgtca
     3361 cgcttacatt cacgccctcc tcccacatcc gctctaaccg aaaaggaagg agttagacaa
     3421 cctgaagtct aggtccctat ttattttttt taatagttat gttagtatta agaacgttat
     3481 ttatatttca aatttttctt ttttttctgt acaaagcggc cgccgaacca ctagtgcact
     3541 gcagtacacg gtttccttga aatttttttg attcggtaat ctccgaacag aaggaagaac
     3601 gaaggaagga gcacagactt agattggtat atatacgcat atgtagtgtt gaagaaacat
     3661 gaaattgccc agtattctta acccaactgc acagaacaaa aacgtgcagg aaacgaagat
     3721 aaatcatgtc gaaagctaca tataaggaac gtgctgctac tcatcctagt cctgttgctg
     3781 ccaagctatt taatatcatg cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc
     3841 gtaccaccaa ggaattactg gagttagttg aagcattagg tcccaaaatt tgtttactaa
     3901 aaacacatgt ggatatcttg actgattttt ccatggaggg cacagttaag ccgctaaagg
     3961 cattatccgc caagtacaat tttttactct tcgaggacag aaaatttgct gacattggta
     4021 atacagtcaa attgcagtac tctgcgggtg tatacagaat agcagaatgg gcagacatta
     4081 cgaatgcaca cggtgtggtg ggcccaggta ttgttagcgg tttgaagcag gcggcagaag
     4141 aagtaacaaa ggaacctaga ggccttttga tgttagcaga attgtcatgc aagggctccc
     4201 tatctactgg agaatatact aagggtactg ttgacattgc gaagagcgac aaagattttg
     4261 ttatcggctt tattgctcaa agagacatgg gtggaagaga tgaaggttac gattggttga
     4321 ttatgacacc cggtgtgggt ttagatgaca agggagatgc attgggtcaa cagtatagaa
     4381 ccgtggatga tgtggtttct acaggatctg acattattat tgttggaaga ggactatttg
     4441 caaagggaag ggatgctaag gtagagggtg aacgttacag aaaagcaggc tgggaagcat
     4501 atttgagaag atgcggccag caaaactaaa aaactgtatt ataagtaaat gcatgtatac
     4561 taaactcaca aattagagct tcaatttaat tatatcagtt attacccgag taacgaagca
     4621 tctgtgcttc attttgtaga acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag
     4681 aatctgagct gcatttttac agaacagaaa tgcaacgcga aagcgctatt ttaccaacga
     4741 agaatctgtg cttcattttt gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa
     4801 caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgagagcg ctattttacc
     4861 aacaaagaat ctatacttct tttttgttct acaaaaatgc atcccgagag cgctattttt
     4921 ctaacaaagc atcttagatt actttttttc tcctttgtgc gctctataat gcagtctctt
     4981 gataactttt tgcactgtag gtccgttaag gttagaagaa ggctactttg gtgtctattt
     5041 tctcttccat aaaaaaagcc tgactccact tcccgcgttt actgattact agcgaagctg
     5101 cgggtgcatt ttttcaagat aaaggcatcc ccgattatat tctataccga tgtggattgc
     5161 gcatactttg tgaacagaaa gtgatagcgt tgatgattct tcattggtca gaaaattatg
     5221 aacggtttct tctattttgt ctctatatac tacgtatagg aaatgtttac attttcgtat
     5281 tgttttcgat tcactctatg aatagttctt actacaattt ttttgtctaa agagtaatac
     5341 tagagataaa cataaaaaat gtagaggtcg agtttagatg caagttcaag gagcgaaagg
     5401 tggatgggta ggttatatag ggatatagca cagagatata tagcaaagag atacttttga
     5461 gcaatgtttg tggaagcggt attcgcaata ttttagtagc tcgttacagt ccggtgcgtt
     5521 tttggttttt tgaaagtgcg tcatcagagc gcttttggtt ttcaaaagcg ctctgaagtt
     5581 cctatacttt ctagctagag aataggaact tcccgagcgg ccgcgtgtta caagcttgac
     5641 tccagcgtaa ctggactgca atcaactcac tggctcacct tcacgggtgg gcctttcttc
     5701 ggtagaaaat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5761 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5821 tttttccgag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     5881 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     5941 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     6001 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     6061 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6121 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6181 aacaggagag cgcacgaggg agcttccagg ggtaaacgcc tggtatcttt atagtcctgt
     6241 cgggtttcgc cacctctgac ttgagcatcg atttttgtga tgctcgtcag gggggcggag
     6301 cctatggaaa aacgccagca acgcagaaag gcccacccga aggtgagcca ggtgattaca
     6361 tttgggccct catcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta
     6421 aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat ccgcgtccag
     6481 ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag
     6541 ggcggttttt tcctgtttgg tcacttacca atgcttaatc agtgaggcac ctatctcagc
     6601 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     6661 gcgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6721 ggctccagat ttatcagcaa taaaccagcc agccgggagg gccgagcgca gaagtgatcc
     6781 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6841 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     6901 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6961 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgccagaag
     7021 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     7081 catgccatcc gtgagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     7141 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     7201 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gtaaactctc
     7261 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     7321 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     7381 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcattttag cttccttagc
     7441 tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg
     7501 aaagttggaa cctcttacgt gccgatcaag tcaaaagcct ccggtcggag gcttttgact
     7561 ttctgctatg gaggtcaggt atgatttaaa tggtcagtta ga
//
