LOCUS       sequence-299929-        8682 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 8682)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 8682)
  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-m4046500
FEATURES             Location/Qualifiers
     source          1..8682
                     /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"
     primer_bind     complement(4437..4455)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             4806..5609
                     /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      5692..6674
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      6752..6781
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      6803..7390
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(7402..7429)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(7453..7644)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(7645..8436,8437..8505))
                     /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     8268..8287
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8506..8608)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(8609..8640)
                     /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 ttctaaaatc
     3301 gaggaaggta agctagtcat ttggataaat ggagataagg gttacaacgg tttggcagaa
     3361 gtaggcaaga aatttgaaaa ggacacaggt atcaaggtga cggttgaaca tccagacaag
     3421 ttagaagaaa aattccctca agttgctgct actggtgatg gtcccgacat tattttctgg
     3481 gcccacgatc gttttggtgg atatgctcag tcaggtttgc tggctgagat caccccagac
     3541 aaggccttcc aagataaatt gtacccattc acttgggatg ctgtcagata caacggtaag
     3601 cttatagcat atccaattgc tgttgaagcc ttatcgttga tctacaataa ggacttgtta
     3661 cctaacccac caaagacatg ggaagaaatt ccagctttgg ataaagagct aaaggccaag
     3721 ggaaaaagcg ctttgatgtt taacttacaa gaaccttatt tcacctggcc attaatcgct
     3781 gctgacggtg gttacgcatt taaatacgaa aacggtaagt atgacattaa ggatgtcggc
     3841 gttgataatg ctggtgcaaa ggctggtttg accttcttgg ttgatttaat aaagaataaa
     3901 cacatgaacg ctgacactga ctactctatc gccgaagccg ctttcaacaa gggtgaaact
     3961 gctatgacca taaatggtcc atgggcctgg agtaacattg atacttctaa agtaaactat
     4021 ggtgtcactg ttttgcctac attcaaaggc caaccatcca agccctttgt cggtgttcta
     4081 tccgctggca tcaatgctgc ttctccaaac aaggagctag ctaaagaatt tttggaaaat
     4141 tacttgttaa ctgacgaagg tttggaggcc gttaacaagg acaagccatt gggtgctgtc
     4201 gctttgaaat cttacgaaga agaattggct aaggatccca gaattgccgc taccatggag
     4261 aacgcccaaa agggtgaaat tatgccaaac atcccacaaa tgtcagcctt ctggtacgca
     4321 gttaggactg ccgtgattaa tgctgcttcc ggtagacaaa ccgtcgacga agcattaaaa
     4381 gatgctcaaa cgtaaacagg ccccttttcc tttgtcgata tcatgtaatt agttatgtca
     4441 cgcttacatt cacgccctcc tcccacatcc gctctaaccg aaaaggaagg agttagacaa
     4501 cctgaagtct aggtccctat ttattttttt taatagttat gttagtatta agaacgttat
     4561 ttatatttca aatttttctt ttttttctgt acaaagcggc cgccgaacca ctagtgcact
     4621 gcagtacacg gtttccttga aatttttttg attcggtaat ctccgaacag aaggaagaac
     4681 gaaggaagga gcacagactt agattggtat atatacgcat atgtagtgtt gaagaaacat
     4741 gaaattgccc agtattctta acccaactgc acagaacaaa aacgtgcagg aaacgaagat
     4801 aaatcatgtc gaaagctaca tataaggaac gtgctgctac tcatcctagt cctgttgctg
     4861 ccaagctatt taatatcatg cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc
     4921 gtaccaccaa ggaattactg gagttagttg aagcattagg tcccaaaatt tgtttactaa
     4981 aaacacatgt ggatatcttg actgattttt ccatggaggg cacagttaag ccgctaaagg
     5041 cattatccgc caagtacaat tttttactct tcgaggacag aaaatttgct gacattggta
     5101 atacagtcaa attgcagtac tctgcgggtg tatacagaat agcagaatgg gcagacatta
     5161 cgaatgcaca cggtgtggtg ggcccaggta ttgttagcgg tttgaagcag gcggcagaag
     5221 aagtaacaaa ggaacctaga ggccttttga tgttagcaga attgtcatgc aagggctccc
     5281 tatctactgg agaatatact aagggtactg ttgacattgc gaagagcgac aaagattttg
     5341 ttatcggctt tattgctcaa agagacatgg gtggaagaga tgaaggttac gattggttga
     5401 ttatgacacc cggtgtgggt ttagatgaca agggagatgc attgggtcaa cagtatagaa
     5461 ccgtggatga tgtggtttct acaggatctg acattattat tgttggaaga ggactatttg
     5521 caaagggaag ggatgctaag gtagagggtg aacgttacag aaaagcaggc tgggaagcat
     5581 atttgagaag atgcggccag caaaactaaa aaactgtatt ataagtaaat gcatgtatac
     5641 taaactcaca aattagagct tcaatttaat tatatcagtt attacccgag taacgaagca
     5701 tctgtgcttc attttgtaga acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag
     5761 aatctgagct gcatttttac agaacagaaa tgcaacgcga aagcgctatt ttaccaacga
     5821 agaatctgtg cttcattttt gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa
     5881 caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgagagcg ctattttacc
     5941 aacaaagaat ctatacttct tttttgttct acaaaaatgc atcccgagag cgctattttt
     6001 ctaacaaagc atcttagatt actttttttc tcctttgtgc gctctataat gcagtctctt
     6061 gataactttt tgcactgtag gtccgttaag gttagaagaa ggctactttg gtgtctattt
     6121 tctcttccat aaaaaaagcc tgactccact tcccgcgttt actgattact agcgaagctg
     6181 cgggtgcatt ttttcaagat aaaggcatcc ccgattatat tctataccga tgtggattgc
     6241 gcatactttg tgaacagaaa gtgatagcgt tgatgattct tcattggtca gaaaattatg
     6301 aacggtttct tctattttgt ctctatatac tacgtatagg aaatgtttac attttcgtat
     6361 tgttttcgat tcactctatg aatagttctt actacaattt ttttgtctaa agagtaatac
     6421 tagagataaa cataaaaaat gtagaggtcg agtttagatg caagttcaag gagcgaaagg
     6481 tggatgggta ggttatatag ggatatagca cagagatata tagcaaagag atacttttga
     6541 gcaatgtttg tggaagcggt attcgcaata ttttagtagc tcgttacagt ccggtgcgtt
     6601 tttggttttt tgaaagtgcg tcatcagagc gcttttggtt ttcaaaagcg ctctgaagtt
     6661 cctatacttt ctagctagag aataggaact tcccgagcgg ccgcgtgtta caagcttgac
     6721 tccagcgtaa ctggactgca atcaactcac tggctcacct tcacgggtgg gcctttcttc
     6781 ggtagaaaat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     6841 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     6901 tttttccgag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     6961 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     7021 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     7081 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     7141 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     7201 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     7261 aacaggagag cgcacgaggg agcttccagg ggtaaacgcc tggtatcttt atagtcctgt
     7321 cgggtttcgc cacctctgac ttgagcatcg atttttgtga tgctcgtcag gggggcggag
     7381 cctatggaaa aacgccagca acgcagaaag gcccacccga aggtgagcca ggtgattaca
     7441 tttgggccct catcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta
     7501 aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat ccgcgtccag
     7561 ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag
     7621 ggcggttttt tcctgtttgg tcacttacca atgcttaatc agtgaggcac ctatctcagc
     7681 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     7741 gcgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     7801 ggctccagat ttatcagcaa taaaccagcc agccgggagg gccgagcgca gaagtgatcc
     7861 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     7921 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     7981 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     8041 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgccagaag
     8101 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     8161 catgccatcc gtgagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     8221 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     8281 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gtaaactctc
     8341 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     8401 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     8461 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcattttag cttccttagc
     8521 tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg
     8581 aaagttggaa cctcttacgt gccgatcaag tcaaaagcct ccggtcggag gcttttgact
     8641 ttctgctatg gaggtcaggt atgatttaaa tggtcagtta ga
//
