LOCUS       sequence-297670-        7650 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 7650)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 7650)
  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-m4046249
FEATURES             Location/Qualifiers
     source          1..7650
                     /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..3360
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(3405..3423)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             3774..4577
                     /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      4660..5642
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      5720..5749
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      5771..6358
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(6370..6397)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(6421..6612)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6613..7404,7405..7473))
                     /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     7236..7255
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7474..7576)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(7577..7608)
                     /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 ttctgattat
     3301 aaagatcatg atggtgatta taaagatcat gatattgatt ataaagatga tgatgataaa
     3361 taaacaggcc ccttttcctt tgtcgatatc atgtaattag ttatgtcacg cttacattca
     3421 cgccctcctc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc tgaagtctag
     3481 gtccctattt atttttttta atagttatgt tagtattaag aacgttattt atatttcaaa
     3541 tttttctttt ttttctgtac aaagcggccg ccgaaccact agtgcactgc agtacacggt
     3601 ttccttgaaa tttttttgat tcggtaatct ccgaacagaa ggaagaacga aggaaggagc
     3661 acagacttag attggtatat atacgcatat gtagtgttga agaaacatga aattgcccag
     3721 tattcttaac ccaactgcac agaacaaaaa cgtgcaggaa acgaagataa atcatgtcga
     3781 aagctacata taaggaacgt gctgctactc atcctagtcc tgttgctgcc aagctattta
     3841 atatcatgca cgaaaagcaa acaaacttgt gtgcttcatt ggatgttcgt accaccaagg
     3901 aattactgga gttagttgaa gcattaggtc ccaaaatttg tttactaaaa acacatgtgg
     3961 atatcttgac tgatttttcc atggagggca cagttaagcc gctaaaggca ttatccgcca
     4021 agtacaattt tttactcttc gaggacagaa aatttgctga cattggtaat acagtcaaat
     4081 tgcagtactc tgcgggtgta tacagaatag cagaatgggc agacattacg aatgcacacg
     4141 gtgtggtggg cccaggtatt gttagcggtt tgaagcaggc ggcagaagaa gtaacaaagg
     4201 aacctagagg ccttttgatg ttagcagaat tgtcatgcaa gggctcccta tctactggag
     4261 aatatactaa gggtactgtt gacattgcga agagcgacaa agattttgtt atcggcttta
     4321 ttgctcaaag agacatgggt ggaagagatg aaggttacga ttggttgatt atgacacccg
     4381 gtgtgggttt agatgacaag ggagatgcat tgggtcaaca gtatagaacc gtggatgatg
     4441 tggtttctac aggatctgac attattattg ttggaagagg actatttgca aagggaaggg
     4501 atgctaaggt agagggtgaa cgttacagaa aagcaggctg ggaagcatat ttgagaagat
     4561 gcggccagca aaactaaaaa actgtattat aagtaaatgc atgtatacta aactcacaaa
     4621 ttagagcttc aatttaatta tatcagttat tacccgagta acgaagcatc tgtgcttcat
     4681 tttgtagaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc
     4741 atttttacag aacagaaatg caacgcgaaa gcgctatttt accaacgaag aatctgtgct
     4801 tcatttttgt aaaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca aagaatctga
     4861 gctgcatttt tacagaacag aaatgcaacg cgagagcgct attttaccaa caaagaatct
     4921 atacttcttt tttgttctac aaaaatgcat cccgagagcg ctatttttct aacaaagcat
     4981 cttagattac tttttttctc ctttgtgcgc tctataatgc agtctcttga taactttttg
     5041 cactgtaggt ccgttaaggt tagaagaagg ctactttggt gtctattttc tcttccataa
     5101 aaaaagcctg actccacttc ccgcgtttac tgattactag cgaagctgcg ggtgcatttt
     5161 ttcaagataa aggcatcccc gattatattc tataccgatg tggattgcgc atactttgtg
     5221 aacagaaagt gatagcgttg atgattcttc attggtcaga aaattatgaa cggtttcttc
     5281 tattttgtct ctatatacta cgtataggaa atgtttacat tttcgtattg ttttcgattc
     5341 actctatgaa tagttcttac tacaattttt ttgtctaaag agtaatacta gagataaaca
     5401 taaaaaatgt agaggtcgag tttagatgca agttcaagga gcgaaaggtg gatgggtagg
     5461 ttatataggg atatagcaca gagatatata gcaaagagat acttttgagc aatgtttgtg
     5521 gaagcggtat tcgcaatatt ttagtagctc gttacagtcc ggtgcgtttt tggttttttg
     5581 aaagtgcgtc atcagagcgc ttttggtttt caaaagcgct ctgaagttcc tatactttct
     5641 agctagagaa taggaacttc ccgagcggcc gcgtgttaca agcttgactc cagcgtaact
     5701 ggactgcaat caactcactg gctcaccttc acgggtgggc ctttcttcgg tagaaaatca
     5761 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     5821 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaggt
     5881 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     5941 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     6001 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     6061 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     6121 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     6181 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     6241 cacgagggag cttccagggg taaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     6301 cctctgactt gagcatcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     6361 cgccagcaac gcagaaaggc ccacccgaag gtgagccagg tgattacatt tgggccctca
     6421 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc
     6481 gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt
     6541 ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc
     6601 ctgtttggtc acttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6661 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatgc gggagggctt
     6721 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6781 atcagcaata aaccagccag ccgggagggc cgagcgcaga agtgatcctg caactttatc
     6841 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6901 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     6961 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     7021 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gccagaagta agttggccgc
     7081 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     7141 gagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     7201 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     7261 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcgt aaactctcaa ggatcttacc
     7321 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     7381 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     7441 aataagggcg acacggaaat gttgaatact cattttagct tccttagctc ctgaaaatct
     7501 cgataactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa agttggaacc
     7561 tcttacgtgc cgatcaagtc aaaagcctcc ggtcggaggc ttttgacttt ctgctatgga
     7621 ggtcaggtat gatttaaatg gtcagttaga
//
