LOCUS       sequence-300389-        7644 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 7644)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 7644)
  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-m4046319
FEATURES             Location/Qualifiers
     source          1..7644
                     /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             3271..3288
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             3289..3354
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(3399..3417)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             3768..4571
                     /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      4654..5636
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      5714..5743
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      5765..6352
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(6364..6391)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(6415..6606)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6607..7398,7399..7467))
                     /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     7230..7249
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7468..7570)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(7571..7602)
                     /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 gctgatgcac aatcgttttt aaaccgggtt tgcggtgtaa gtgcagcccg tcttacaccg
      541 tgcggcacag gcactagtac tgatgtcgta tacagggctt ttgacatcta caatgataaa
      601 gtagctggtt ttgctaaatt cctaaaaact aattgttgtc gcttccaaga aaaggacgaa
      661 gatgacaatt taattgattc ttactttgta gttaagagac acactttctc taactaccaa
      721 catgaagaaa caatttataa tttacttaag gattgtccag ctgttgctaa acatgacttc
      781 tttaagttta gaatagacgg tgacatggta ccacatatat cacgtcaacg tcttactaaa
      841 tacacaatgg cagacctcgt ctatgcttta aggcattttg atgaaggtaa ttgtgacaca
      901 ttaaaagaaa tacttgtcac atacaattgt tgtgatgatg attatttcaa taaaaaggac
      961 tggtatgatt ttgtagaaaa cccagatata ttacgcgtat acgccaactt aggtgaacgt
     1021 gtacgccaag ctttgttaaa aacagtacaa ttctgtgatg ccatgcgaaa tgctggtatt
     1081 gttggtgtac tgacattaga taatcaagat ctcaatggta actggtatga tttcggtgat
     1141 ttcatacaaa ccacgccagg tagtggagtt cctgttgtag attcttatta ttcattgtta
     1201 atgcctatat taaccttgac cagggcttta actgcagagt cacatgttga cactgactta
     1261 acaaagcctt acattaagtg ggatttgtta aaatatgact tcacggaaga gaggttaaaa
     1321 ctctttgacc gttattttaa atattgggat cagacatacc acccaaattg tgttaactgt
     1381 ttggatgaca gatgcattct gcattgtgca aactttaatg ttttattctc tacagtgttc
     1441 ccacctacaa gttttggacc actagtgaga aaaatatttg ttgatggtgt tccatttgta
     1501 gtttcaactg gataccactt cagagagcta ggtgttgtac ataatcagga tgtaaactta
     1561 catagctcta gacttagttt taaggaatta cttgtgtatg ctgctgaccc tgctatgcac
     1621 gctgcttctg gtaatctatt actagataaa cgcactacgt gcttttcagt agctgcactt
     1681 actaacaatg ttgcttttca aactgtcaaa cccggtaatt ttaacaaaga cttctatgac
     1741 tttgctgtgt ctaagggttt ctttaaggaa ggaagttctg ttgaattaaa acacttcttc
     1801 tttgctcagg atggtaatgc tgctatcagc gattatgact actatcgtta taatctacca
     1861 acaatgtgtg atatcagaca actactattt gtagttgaag ttgttgataa gtactttgat
     1921 tgttacgatg gtggctgtat taatgctaac caagtcatcg tcaacaacct agacaaatca
     1981 gctggttttc catttaataa atggggtaag gctagacttt attatgattc aatgagttat
     2041 gaggatcaag atgcactttt cgcatataca aaacgtaatg tcatccctac tataactcaa
     2101 atgaatctta agtatgccat tagtgcaaag aatagagctc gcaccgtagc tggtgtctct
     2161 atctgtagta ctatgaccaa tagacagttt catcaaaaat tattgaaatc aatagccgcc
     2221 actagaggag ctactgtagt aattggaaca agcaaattct atggtggttg gcacaacatg
     2281 ttaaaaactg tttatagtga tgtagaaaac cctcacctta tgggttggga ttatcctaaa
     2341 tgtgatagag ccatgcctaa catgcttaga attatggcct cacttgttct tgctcgcaaa
     2401 catacaacgt gttgtagctt gtcacaccgt ttctatagat tagctaatga gtgtgctcaa
     2461 gtattgagtg aaatggtcat gtgtggcggt tcactatatg ttaaaccagg tggaacctca
     2521 tcaggagatg ccacaactgc ttatgctaat agtgttttta acatttgtca agctgtcacg
     2581 gccaatgtta atgcactttt atctactgat ggtaacaaaa ttgccgataa gtatgtccgc
     2641 aatttacaac acagacttta tgagtgtctc tatagaaata gagatgttga cacagacttt
     2701 gtgaatgagt tttacgcata tttgcgtaaa catttctcaa tgatgatact ctctgacgat
     2761 gctgttgtgt gtttcaatag cacttatgca tctcaaggtc tagtggctag cataaagaac
     2821 tttaagtcag ttctttatta tcaaaacaat gtttttatgt ctgaagcaaa atgttggact
     2881 gagactgacc ttactaaagg acctcatgaa ttttgctctc aacatacaat gctagttaaa
     2941 cagggtgatg attatgtgta ccttccttac ccagatccat caagaatcct aggggccggc
     3001 tgttttgtag atgatatcgt aaaaacagat ggtacactta tgattgaacg gttcgtgtct
     3061 ttagctatag atgcttaccc acttactaaa catcctaatc aggagtatgc tgatgtcttt
     3121 catttgtact tacaatacat aagaaagcta catgatgagt taacaggaca catgttagac
     3181 atgtattctg ttatgcttac taatgataac acttcaaggt attgggaacc tgagttttat
     3241 gaggctatgt acacaccgca tacagtctta ctcgtgccga gaggttctga ttataaagat
     3301 catgatggtg attataaaga tcatgatatt gattataaag atgatgatga taaataaaca
     3361 ggcccctttt cctttgtcga tatcatgtaa ttagttatgt cacgcttaca ttcacgccct
     3421 cctcccacat ccgctctaac cgaaaaggaa ggagttagac aacctgaagt ctaggtccct
     3481 atttattttt tttaatagtt atgttagtat taagaacgtt atttatattt caaatttttc
     3541 ttttttttct gtacaaagcg gccgccgaac cactagtgca ctgcagtaca cggtttcctt
     3601 gaaatttttt tgattcggta atctccgaac agaaggaaga acgaaggaag gagcacagac
     3661 ttagattggt atatatacgc atatgtagtg ttgaagaaac atgaaattgc ccagtattct
     3721 taacccaact gcacagaaca aaaacgtgca ggaaacgaag ataaatcatg tcgaaagcta
     3781 catataagga acgtgctgct actcatccta gtcctgttgc tgccaagcta tttaatatca
     3841 tgcacgaaaa gcaaacaaac ttgtgtgctt cattggatgt tcgtaccacc aaggaattac
     3901 tggagttagt tgaagcatta ggtcccaaaa tttgtttact aaaaacacat gtggatatct
     3961 tgactgattt ttccatggag ggcacagtta agccgctaaa ggcattatcc gccaagtaca
     4021 attttttact cttcgaggac agaaaatttg ctgacattgg taatacagtc aaattgcagt
     4081 actctgcggg tgtatacaga atagcagaat gggcagacat tacgaatgca cacggtgtgg
     4141 tgggcccagg tattgttagc ggtttgaagc aggcggcaga agaagtaaca aaggaaccta
     4201 gaggcctttt gatgttagca gaattgtcat gcaagggctc cctatctact ggagaatata
     4261 ctaagggtac tgttgacatt gcgaagagcg acaaagattt tgttatcggc tttattgctc
     4321 aaagagacat gggtggaaga gatgaaggtt acgattggtt gattatgaca cccggtgtgg
     4381 gtttagatga caagggagat gcattgggtc aacagtatag aaccgtggat gatgtggttt
     4441 ctacaggatc tgacattatt attgttggaa gaggactatt tgcaaaggga agggatgcta
     4501 aggtagaggg tgaacgttac agaaaagcag gctgggaagc atatttgaga agatgcggcc
     4561 agcaaaacta aaaaactgta ttataagtaa atgcatgtat actaaactca caaattagag
     4621 cttcaattta attatatcag ttattacccg agtaacgaag catctgtgct tcattttgta
     4681 gaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt
     4741 acagaacaga aatgcaacgc gaaagcgcta ttttaccaac gaagaatctg tgcttcattt
     4801 ttgtaaaaca aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca
     4861 tttttacaga acagaaatgc aacgcgagag cgctatttta ccaacaaaga atctatactt
     4921 cttttttgtt ctacaaaaat gcatcccgag agcgctattt ttctaacaaa gcatcttaga
     4981 ttactttttt tctcctttgt gcgctctata atgcagtctc ttgataactt tttgcactgt
     5041 aggtccgtta aggttagaag aaggctactt tggtgtctat tttctcttcc ataaaaaaag
     5101 cctgactcca cttcccgcgt ttactgatta ctagcgaagc tgcgggtgca ttttttcaag
     5161 ataaaggcat ccccgattat attctatacc gatgtggatt gcgcatactt tgtgaacaga
     5221 aagtgatagc gttgatgatt cttcattggt cagaaaatta tgaacggttt cttctatttt
     5281 gtctctatat actacgtata ggaaatgttt acattttcgt attgttttcg attcactcta
     5341 tgaatagttc ttactacaat ttttttgtct aaagagtaat actagagata aacataaaaa
     5401 atgtagaggt cgagtttaga tgcaagttca aggagcgaaa ggtggatggg taggttatat
     5461 agggatatag cacagagata tatagcaaag agatactttt gagcaatgtt tgtggaagcg
     5521 gtattcgcaa tattttagta gctcgttaca gtccggtgcg tttttggttt tttgaaagtg
     5581 cgtcatcaga gcgcttttgg ttttcaaaag cgctctgaag ttcctatact ttctagctag
     5641 agaataggaa cttcccgagc ggccgcgtgt tacaagcttg actccagcgt aactggactg
     5701 caatcaactc actggctcac cttcacgggt gggcctttct tcggtagaaa atcaaaggat
     5761 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5821 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aggtaactgg
     5881 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     5941 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     6001 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     6061 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     6121 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     6181 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     6241 ggagcttcca ggggtaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     6301 acttgagcat cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     6361 caacgcagaa aggcccaccc gaaggtgagc caggtgatta catttgggcc ctcatcagag
     6421 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
     6481 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
     6541 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
     6601 ggtcacttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6661 atccatagtt gcctgactcc ccgtcgtgta gataactacg atgcgggagg gcttaccatc
     6721 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6781 aataaaccag ccagccggga gggccgagcg cagaagtgat cctgcaactt tatccgcctc
     6841 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6901 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6961 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     7021 aaaagcggtt agctccttcg gtcctccgat cgttgccaga agtaagttgg ccgcagtgtt
     7081 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtgagatg
     7141 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     7201 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     7261 agtgctcatc attggaaaac gttcttcggg gcgtaaactc tcaaggatct taccgctgtt
     7321 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7381 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7441 ggcgacacgg aaatgttgaa tactcatttt agcttcctta gctcctgaaa atctcgataa
     7501 ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg aacctcttac
     7561 gtgccgatca agtcaaaagc ctccggtcgg aggcttttga ctttctgcta tggaggtcag
     7621 gtatgattta aatggtcagt taga
//
