LOCUS       sequence-297547-        7566 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 7566)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 7566)
  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-m4046670
FEATURES             Location/Qualifiers
     source          1..7566
                     /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"
     primer_bind     complement(3321..3339)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             3690..4493
                     /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      4576..5558
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      5636..5665
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      5687..6274
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(6286..6313)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(6337..6528)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6529..7320,7321..7389))
                     /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     7152..7171
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7390..7492)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(7493..7524)
                     /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 ttgcagtaaa caggcccctt ttcctttgtc
     3301 gatatcatgt aattagttat gtcacgctta cattcacgcc ctcctcccac atccgctcta
     3361 accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt tttttaatag
     3421 ttatgttagt attaagaacg ttatttatat ttcaaatttt tctttttttt ctgtacaaag
     3481 cggccgccga accactagtg cactgcagta cacggtttcc ttgaaatttt tttgattcgg
     3541 taatctccga acagaaggaa gaacgaagga aggagcacag acttagattg gtatatatac
     3601 gcatatgtag tgttgaagaa acatgaaatt gcccagtatt cttaacccaa ctgcacagaa
     3661 caaaaacgtg caggaaacga agataaatca tgtcgaaagc tacatataag gaacgtgctg
     3721 ctactcatcc tagtcctgtt gctgccaagc tatttaatat catgcacgaa aagcaaacaa
     3781 acttgtgtgc ttcattggat gttcgtacca ccaaggaatt actggagtta gttgaagcat
     3841 taggtcccaa aatttgttta ctaaaaacac atgtggatat cttgactgat ttttccatgg
     3901 agggcacagt taagccgcta aaggcattat ccgccaagta caatttttta ctcttcgagg
     3961 acagaaaatt tgctgacatt ggtaatacag tcaaattgca gtactctgcg ggtgtataca
     4021 gaatagcaga atgggcagac attacgaatg cacacggtgt ggtgggccca ggtattgtta
     4081 gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc tagaggcctt ttgatgttag
     4141 cagaattgtc atgcaagggc tccctatcta ctggagaata tactaagggt actgttgaca
     4201 ttgcgaagag cgacaaagat tttgttatcg gctttattgc tcaaagagac atgggtggaa
     4261 gagatgaagg ttacgattgg ttgattatga cacccggtgt gggtttagat gacaagggag
     4321 atgcattggg tcaacagtat agaaccgtgg atgatgtggt ttctacagga tctgacatta
     4381 ttattgttgg aagaggacta tttgcaaagg gaagggatgc taaggtagag ggtgaacgtt
     4441 acagaaaagc aggctgggaa gcatatttga gaagatgcgg ccagcaaaac taaaaaactg
     4501 tattataagt aaatgcatgt atactaaact cacaaattag agcttcaatt taattatatc
     4561 agttattacc cgagtaacga agcatctgtg cttcattttg tagaacaaaa atgcaacgcg
     4621 agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac
     4681 gcgaaagcgc tattttacca acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca
     4741 acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat
     4801 gcaacgcgag agcgctattt taccaacaaa gaatctatac ttcttttttg ttctacaaaa
     4861 atgcatcccg agagcgctat ttttctaaca aagcatctta gattactttt tttctccttt
     4921 gtgcgctcta taatgcagtc tcttgataac tttttgcact gtaggtccgt taaggttaga
     4981 agaaggctac tttggtgtct attttctctt ccataaaaaa agcctgactc cacttcccgc
     5041 gtttactgat tactagcgaa gctgcgggtg cattttttca agataaaggc atccccgatt
     5101 atattctata ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata gcgttgatga
     5161 ttcttcattg gtcagaaaat tatgaacggt ttcttctatt ttgtctctat atactacgta
     5221 taggaaatgt ttacattttc gtattgtttt cgattcactc tatgaatagt tcttactaca
     5281 atttttttgt ctaaagagta atactagaga taaacataaa aaatgtagag gtcgagttta
     5341 gatgcaagtt caaggagcga aaggtggatg ggtaggttat atagggatat agcacagaga
     5401 tatatagcaa agagatactt ttgagcaatg tttgtggaag cggtattcgc aatattttag
     5461 tagctcgtta cagtccggtg cgtttttggt tttttgaaag tgcgtcatca gagcgctttt
     5521 ggttttcaaa agcgctctga agttcctata ctttctagct agagaatagg aacttcccga
     5581 gcggccgcgt gttacaagct tgactccagc gtaactggac tgcaatcaac tcactggctc
     5641 accttcacgg gtgggccttt cttcggtaga aaatcaaagg atcttcttga gatccttttt
     5701 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5761 tgccggatca agagctacca actctttttc cgaggtaact ggcttcagca gagcgcagat
     5821 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5881 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5941 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     6001 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     6061 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     6121 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc caggggtaaa
     6181 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc atcgattttt
     6241 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcag aaaggcccac
     6301 ccgaaggtga gccaggtgat tacatttggg ccctcatcag aggttttcac cgtcatcacc
     6361 gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat
     6421 gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct
     6481 tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactt accaatgctt
     6541 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     6601 ccccgtcgtg tagataacta cgatgcggga gggcttacca tctggcccca gtgctgcaat
     6661 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     6721 gagggccgag cgcagaagtg atcctgcaac tttatccgcc tccatccagt ctattaattg
     6781 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     6841 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     6901 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     6961 cggtcctccg atcgttgcca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     7021 agcactgcat aattctctta ctgtcatgcc atccgtgaga tgcttttctg tgactggtga
     7081 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     7141 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     7201 acgttcttcg gggcgtaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     7261 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     7321 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     7381 aatactcatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa atacgcccgg
     7441 tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat caagtcaaaa
     7501 gcctccggtc ggaggctttt gactttctgc tatggaggtc aggtatgatt taaatggtca
     7561 gttaga
//
