LOCUS       sequence-298238-        7596 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 7596)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 7596)
  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-m4046259
FEATURES             Location/Qualifiers
     source          1..7596
                     /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..3306
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3351..3369)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             3720..4523
                     /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      4606..5588
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      5666..5695
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      5717..6304
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(6316..6343)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(6367..6558)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6559..7350,7351..7419))
                     /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     7182..7201
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7420..7522)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(7523..7554)
                     /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 gaggttctca ccatcatcac
     3301 catcattaaa caggcccctt ttcctttgtc gatatcatgt aattagttat gtcacgctta
     3361 cattcacgcc ctcctcccac atccgctcta accgaaaagg aaggagttag acaacctgaa
     3421 gtctaggtcc ctatttattt tttttaatag ttatgttagt attaagaacg ttatttatat
     3481 ttcaaatttt tctttttttt ctgtacaaag cggccgccga accactagtg cactgcagta
     3541 cacggtttcc ttgaaatttt tttgattcgg taatctccga acagaaggaa gaacgaagga
     3601 aggagcacag acttagattg gtatatatac gcatatgtag tgttgaagaa acatgaaatt
     3661 gcccagtatt cttaacccaa ctgcacagaa caaaaacgtg caggaaacga agataaatca
     3721 tgtcgaaagc tacatataag gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc
     3781 tatttaatat catgcacgaa aagcaaacaa acttgtgtgc ttcattggat gttcgtacca
     3841 ccaaggaatt actggagtta gttgaagcat taggtcccaa aatttgttta ctaaaaacac
     3901 atgtggatat cttgactgat ttttccatgg agggcacagt taagccgcta aaggcattat
     3961 ccgccaagta caatttttta ctcttcgagg acagaaaatt tgctgacatt ggtaatacag
     4021 tcaaattgca gtactctgcg ggtgtataca gaatagcaga atgggcagac attacgaatg
     4081 cacacggtgt ggtgggccca ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa
     4141 caaaggaacc tagaggcctt ttgatgttag cagaattgtc atgcaagggc tccctatcta
     4201 ctggagaata tactaagggt actgttgaca ttgcgaagag cgacaaagat tttgttatcg
     4261 gctttattgc tcaaagagac atgggtggaa gagatgaagg ttacgattgg ttgattatga
     4321 cacccggtgt gggtttagat gacaagggag atgcattggg tcaacagtat agaaccgtgg
     4381 atgatgtggt ttctacagga tctgacatta ttattgttgg aagaggacta tttgcaaagg
     4441 gaagggatgc taaggtagag ggtgaacgtt acagaaaagc aggctgggaa gcatatttga
     4501 gaagatgcgg ccagcaaaac taaaaaactg tattataagt aaatgcatgt atactaaact
     4561 cacaaattag agcttcaatt taattatatc agttattacc cgagtaacga agcatctgtg
     4621 cttcattttg tagaacaaaa atgcaacgcg agagcgctaa tttttcaaac aaagaatctg
     4681 agctgcattt ttacagaaca gaaatgcaac gcgaaagcgc tattttacca acgaagaatc
     4741 tgtgcttcat ttttgtaaaa caaaaatgca acgcgagagc gctaattttt caaacaaaga
     4801 atctgagctg catttttaca gaacagaaat gcaacgcgag agcgctattt taccaacaaa
     4861 gaatctatac ttcttttttg ttctacaaaa atgcatcccg agagcgctat ttttntaaca
     4921 aagcatctta gattactttt tttctccttt gtgcgctcta taatgcagtc tcttgataac
     4981 tttttgcact gtaggtccgt taaggttaga agaaggctac tttggtgtct attttctctt
     5041 ccataaaaaa agcctgactc cacttcccgc gtttactgat tactagcgaa gctgcgggtg
     5101 cattttttca agataaaggc atccccgatt atattctata ccgatgtgga ttgcgcatac
     5161 tttgtgaaca gaaagtgata gcgttgatga ttcttcattg gtcagaaaat tatgaacggt
     5221 ttcttctatt ttgtctctat atactacgta taggaaatgt ttacattttc gtattgtttt
     5281 cgattcactc tatgaatagt tcttactaca atttttttgt ctaaagagta atactagaga
     5341 taaacataaa aaatgtagag gtcgagttta gatgcaagtt caaggagcga aaggtggatg
     5401 ggtaggttat atagggatat agcacagaga tatatagcaa agagatactt ttgagcaatg
     5461 tttgtggaag cggtattcgc aatattttag tagctcgtta cagtccggtg cgtttttggt
     5521 tttttgaaag tgcgtcatca gagcgctttt ggttttcaaa agcgctctga agttcctata
     5581 ctttctagct agagaatagg aacttcccga gcggccgcgt gttacaagct tgactccagc
     5641 gtaactggac tgcaatcaac tcactggctc accttcacgg gtgggccttt cttcggtaga
     5701 aaatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     5761 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     5821 cgaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     5881 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5941 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6001 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6061 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6121 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6181 agagcgcacg agggagcttc caggggtaaa cgcctggtat ctttatagtc ctgtcgggtt
     6241 tcgccacctc tgacttgagc atcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6301 gaaaaacgcc agcaacgcag aaaggcccac ccgaaggtga gccaggtgat tacatttggg
     6361 ccctcatcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc
     6421 atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt
     6481 gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt
     6541 tttttcctgt ttggtcactt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6601 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatgcggga
     6661 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6721 agatttatca gcaataaacc agccagccgg gagggccgag cgcagaagtg atcctgcaac
     6781 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6841 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6901 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6961 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgcca gaagtaagtt
     7021 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     7081 atccgtgaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     7141 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     7201 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgtaaac tctcaaggat
     7261 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     7321 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     7381 aaagggaata agggcgacac ggaaatgttg aatactcatt ttagcttcct tagctcctga
     7441 aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt
     7501 ggaacctctt acgtgccgat caagtcaaaa gcctccggtc ggaggctttt gactttctgc
     7561 tatggaggtc aggtatgatt taaatggtca gttaga
//
