LOCUS       sequence-291479-        8676 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 8676)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 8676)
  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-m4046514
FEATURES             Location/Qualifiers
     source          1..8676
                     /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"
     primer_bind     complement(4431..4449)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             4800..5603
                     /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      5686..6668
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      6746..6775
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      6797..7384
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(7396..7423)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(7447..7638)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(7639..8430,8431..8499))
                     /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     8262..8281
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8500..8602)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(8603..8634)
                     /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 gaggttctaa aatcgaggaa
     3301 ggtaagctag tcatttggat aaatggagat aagggttaca acggtttggc agaagtaggc
     3361 aagaaatttg aaaaggacac aggtatcaag gtgacggttg aacatccaga caagttagaa
     3421 gaaaaattcc ctcaagttgc tgctactggt gatggtcccg acattatttt ctgggcccac
     3481 gatcgttttg gtggatatgc tcagtcaggt ttgctggctg agatcacccc agacaaggcc
     3541 ttccaagata aattgtaccc attcacttgg gatgctgtca gatacaacgg taagcttata
     3601 gcatatccaa ttgctgttga agccttatcg ttgatctaca ataaggactt gttacctaac
     3661 ccaccaaaga catgggaaga aattccagct ttggataaag agctaaaggc caagggaaaa
     3721 agcgctttga tgtttaactt acaagaacct tatttcacct ggccattaat cgctgctgac
     3781 ggtggttacg catttaaata cgaaaacggt aagtatgaca ttaaggatgt cggcgttgat
     3841 aatgctggtg caaaggctgg tttgaccttc ttggttgatt taataaagaa taaacacatg
     3901 aacgctgaca ctgactactc tatcgccgaa gccgctttca acaagggtga aactgctatg
     3961 accataaatg gtccatgggc ctggagtaac attgatactt ctaaagtaaa ctatggtgtc
     4021 actgttttgc ctacattcaa aggccaacca tccaagccct ttgtcggtgt tctatccgct
     4081 ggcatcaatg ctgcttctcc aaacaaggag ctagctaaag aatttttgga aaattacttg
     4141 ttaactgacg aaggtttgga ggccgttaac aaggacaagc cattgggtgc tgtcgctttg
     4201 aaatcttacg aagaagaatt ggctaaggat cccagaattg ccgctaccat ggagaacgcc
     4261 caaaagggtg aaattatgcc aaacatccca caaatgtcag ccttctggta cgcagttagg
     4321 actgccgtga ttaatgctgc ttccggtaga caaaccgtcg acgaagcatt aaaagatgct
     4381 caaacgtaaa caggcccctt ttcctttgtc gatatcatgt aattagttat gtcacgctta
     4441 cattcacgcc ctcctcccac atccgctcta accgaaaagg aaggagttag acaacctgaa
     4501 gtctaggtcc ctatttattt tttttaatag ttatgttagt attaagaacg ttatttatat
     4561 ttcaaatttt tctttttttt ctgtacaaag cggccgccga accactagtg cactgcagta
     4621 cacggtttcc ttgaaatttt tttgattcgg taatctccga acagaaggaa gaacgaagga
     4681 aggagcacag acttagattg gtatatatac gcatatgtag tgttgaagaa acatgaaatt
     4741 gcccagtatt cttaacccaa ctgcacagaa caaaaacgtg caggaaacga agataaatca
     4801 tgtcgaaagc tacatataag gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc
     4861 tatttaatat catgcacgaa aagcaaacaa acttgtgtgc ttcattggat gttcgtacca
     4921 ccaaggaatt actggagtta gttgaagcat taggtcccaa aatttgttta ctaaaaacac
     4981 atgtggatat cttgactgat ttttccatgg agggcacagt taagccgcta aaggcattat
     5041 ccgccaagta caatttttta ctcttcgagg acagaaaatt tgctgacatt ggtaatacag
     5101 tcaaattgca gtactctgcg ggtgtataca gaatagcaga atgggcagac attacgaatg
     5161 cacacggtgt ggtgggccca ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa
     5221 caaaggaacc tagaggcctt ttgatgttag cagaattgtc atgcaagggc tccctatcta
     5281 ctggagaata tactaagggt actgttgaca ttgcgaagag cgacaaagat tttgttatcg
     5341 gctttattgc tcaaagagac atgggtggaa gagatgaagg ttacgattgg ttgattatga
     5401 cacccggtgt gggtttagat gacaagggag atgcattggg tcaacagtat agaaccgtgg
     5461 atgatgtggt ttctacagga tctgacatta ttattgttgg aagaggacta tttgcaaagg
     5521 gaagggatgc taaggtagag ggtgaacgtt acagaaaagc aggctgggaa gcatatttga
     5581 gaagatgcgg ccagcaaaac taaaaaactg tattataagt aaatgcatgt atactaaact
     5641 cacaaattag agcttcaatt taattatatc agttattacc cgagtaacga agcatctgtg
     5701 cttcattttg tagaacaaaa atgcaacgcg agagcgctaa tttttcaaac aaagaatctg
     5761 agctgcattt ttacagaaca gaaatgcaac gcgaaagcgc tattttacca acgaagaatc
     5821 tgtgcttcat ttttgtaaaa caaaaatgca acgcgagagc gctaattttt caaacaaaga
     5881 atctgagctg catttttaca gaacagaaat gcaacgcgag agcgctattt taccaacaaa
     5941 gaatctatac ttcttttttg ttctacaaaa atgcatcccg agagcgctat ttttctaaca
     6001 aagcatctta gattactttt tttctccttt gtgcgctcta taatgcagtc tcttgataac
     6061 tttttgcact gtaggtccgt taaggttaga agaaggctac tttggtgtct attttctctt
     6121 ccataaaaaa agcctgactc cacttcccgc gtttactgat tactagcgaa gctgcgggtg
     6181 cattttttca agataaaggc atccccgatt atattctata ccgatgtgga ttgcgcatac
     6241 tttgtgaaca gaaagtgata gcgttgatga ttcttcattg gtcagaaaat tatgaacggt
     6301 ttcttctatt ttgtctctat atactacgta taggaaatgt ttacattttc gtattgtttt
     6361 cgattcactc tatgaatagt tcttactaca atttttttgt ctaaagagta atactagaga
     6421 taaacataaa aaatgtagag gtcgagttta gatgcaagtt caaggagcga aaggtggatg
     6481 ggtaggttat atagggatat agcacagaga tatatagcaa agagatactt ttgagcaatg
     6541 tttgtggaag cggtattcgc aatattttag tagctcgtta cagtccggtg cgtttttggt
     6601 tttttgaaag tgcgtcatca gagcgctttt ggttttcaaa agcgctctga agttcctata
     6661 ctttctagct agagaatagg aacttcccga gcggccgcgt gttacaagct tgactccagc
     6721 gtaactggac tgcaatcaac tcactggctc accttcacgg gtgggccttt cttcggtaga
     6781 aaatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6841 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6901 cgaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     6961 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     7021 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     7081 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     7141 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     7201 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     7261 agagcgcacg agggagcttc caggggtaaa cgcctggtat ctttatagtc ctgtcgggtt
     7321 tcgccacctc tgacttgagc atcgattttt gtgatgctcg tcaggggggc ggagcctatg
     7381 gaaaaacgcc agcaacgcag aaaggcccac ccgaaggtga gccaggtgat tacatttggg
     7441 ccctcatcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc
     7501 atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt
     7561 gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt
     7621 tttttcctgt ttggtcactt accaatgctt aatcagtgag gcacctatct cagcgatctg
     7681 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatgcggga
     7741 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     7801 agatttatca gcaataaacc agccagccgg gagggccgag cgcagaagtg atcctgcaac
     7861 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     7921 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     7981 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     8041 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgcca gaagtaagtt
     8101 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     8161 atccgtgaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     8221 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     8281 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgtaaac tctcaaggat
     8341 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     8401 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     8461 aaagggaata agggcgacac ggaaatgttg aatactcatt ttagcttcct tagctcctga
     8521 aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt
     8581 ggaacctctt acgtgccgat caagtcaaaa gcctccggtc ggaggctttt gactttctgc
     8641 tatggaggtc aggtatgatt taaatggtca gttaga
//
