LOCUS       sequence-297582-        7519 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 7519)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 7519)
  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-m4046618
FEATURES             Location/Qualifiers
     source          1..7519
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     complement(3274..3292)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             3643..4446
                     /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      4529..5511
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      5589..5618
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      5640..6227
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(6239..6266)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(6290..6481)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6482..7273,7274..7342))
                     /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     7105..7124
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7343..7445)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(7446..7477)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgccc cacacaccat agcttcaaaa tgtttctact ccttttttac tcttccagat
       61 tttctcggac tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag catactaaat
      121 ttcccctctt tcttcctcta gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa
      181 aaaagagacc gcctcgtttc tttttcttcg tcgaaaaagg caataaaaat ttttatcacg
      241 tttctttttc ttgaaaattt ttttttttga tttttttctc tttcgatgac ctcccattga
      301 tatttaagtt aataaacggt cttcaatttc tcaagtttca gtttcatttt tcttgttcta
      361 ttacaacttt ttttacttct tgctcattag aaagaaagca tagcaatcta atctaagttt
      421 taattacaaa atgtccgccg atgcacaatc tttcttaaat agagtctgcg gcgtgagtgc
      481 tgctcgtcta accccttgtg gtacgggtac ttcaacagac gtagtttaca gggcgtttga
      541 tatttataac gacaaggtcg ctggtttcgc taaatttttg aagaccaact gttgtagatt
      601 ccaggagaag gatgaagacg acaatcttat cgattcgtac ttcgttgtta aaagacatac
      661 tttttccaac tatcaacacg aagaaacaat atacaatttg ctcaaggact gtccagccgt
      721 cgctaaacac gattttttca agttcagaat tgatggggac atggtgccac atatcagcag
      781 acaacggttg accaagtaca ctatggctga tttagtttac gccttgcgac actttgacga
      841 aggaaactgt gatactctga aggaaatttt ggttacctat aactgctgtg acgatgatta
      901 cttcaacaaa aaggactggt acgacttcgt agagaatccc gacatcttga gggtttacgc
      961 taacttaggt gaaagagtca gacaagcttt gttaaagaca gtccaatttt gcgatgctat
     1021 gagaaatgcg ggtattgttg gtgttttgac tttggataat caagatttga acggtaactg
     1081 gtatgacttc ggtgatttta tccagactac cccgggttct ggggtaccag ttgtcgattc
     1141 ttactattct ctgttaatgc caatactcac cctaactaga gcattgactg ccgaatccca
     1201 tgttgacact gatttaacta agccatacat taaatgggac cttctaaagt acgatttcac
     1261 cgaagaacgt ttaaagttgt tcgacagata ttttaaatac tgggaccaaa cctaccaccc
     1321 taattgtgtc aactgtttgg acgacagatg tattcttcac tgcgctaact ttaatgtgtt
     1381 gttctcgact gtttttccac caacctcttt cggtccgctg gttcgaaaga tttttgttga
     1441 cggtgttcca ttcgtggtca gtacgggata tcatttcaga gaattagggg tcgttcataa
     1501 ccaagacgtg aacttgcact catcaagatt gtccttcaag gaattgttgg tctacgcagc
     1561 tgatcctgca atgcacgccg cttctggaaa cttattgctg gataagagaa caacctgttt
     1621 cagcgttgcc gctcttacta ataacgtcgc gttccaaact gtaaagccgg gtaactttaa
     1681 taaagacttc tacgattttg ctgttagcaa gggttttttc aaagaaggtt ctagtgttga
     1741 gttgaaacac tttttctttg ctcaagacgg caatgctgct atcagtgatt atgactatta
     1801 ccgttacaac ttaccaacaa tgtgtgatat cagacagttg cttttcgtcg tcgaagttgt
     1861 tgataaatat tttgactgtt atgatggtgg ttgcatcaat gcaaatcaag tcattgtcaa
     1921 caacttagat aagtctgctg gcttcccatt caacaagtgg ggtaaagcca gattgtacta
     1981 tgattccatg tcctacgagg accaagatgc tttgttcgcg tacaccaaga gaaacgttat
     2041 tcctacaatt actcaaatga atctaaagta cgctatctct gcaaaaaatc gggccagaac
     2101 cgttgctggt gtttctatct gttctacgat gactaaccgt caatttcatc agaagctatt
     2161 gaaatccata gctgctacta gaggtgctac tgtagtcatc ggtacctcaa agttctatgg
     2221 tggttggcac aatatgctca agaccgttta ctctgatgtc gaaaacccac atcttatggg
     2281 ttgggactac cctaagtgtg atagagccat gccaaatatg ctaagaatta tggcatcctt
     2341 ggtcttggct agaaagcata ccacatgctg ctcattgtcc caccgttttt acagattggc
     2401 caacgaatgt gctcaagtcc tttctgaaat ggttatgtgt ggtggtagtc tatatgtaaa
     2461 gccaggcgga acttcgtctg gagatgccac cactgcttac gctaactcgg tttttaacat
     2521 ttgtcaagcc gttaccgcca acgtaaacgc tttgctgtct acggatggta acaagatcgc
     2581 tgataagtat gtgcgcaact tgcaacatag actttatgaa tgtttgtata gaaatcgaga
     2641 tgtcgatact gacttcgtta acgaatttta cgcgtactta agaaagcact tctccatgat
     2701 gattttgtca gacgacgctg ttgtttgttt caattctact tacgctagcc aaggtttggt
     2761 cgcttcaatc aaaaacttca agtctgtttt atactaccaa aacaatgttt ttatgtccga
     2821 ggcaaaatgc tggacggaaa cagatttgac taagggtcca catgaatttt gtagtcaaca
     2881 cacaatgtta gtgaaacaag gggacgacta cgtttaccta ccctacccag atccaagcag
     2941 aattttgggt gccggttgtt tcgtggacga catagtcaag accgacggta ccttgatgat
     3001 cgaacggttt gtctctctgg ccatagacgc ttacccattg actaagcacc ctaaccaaga
     3061 atatgctgat gtttttcatt tatatcttca atacattaga aaattacatg acgaattgac
     3121 cggtcacatg ttagatatgt actctgtgat gttgacgaac gataatacgt cacgttactg
     3181 ggaacccgaa ttttacgaag cgatgtacac tccacacaca gtattgcagt aaacaggccc
     3241 cttttccttt gtcgatatca tgtaattagt tatgtcacgc ttacattcac gccctcctcc
     3301 cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta
     3361 ttttttttaa tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt
     3421 tttctgtaca aagcggccgc cgaaccacta gtgcactgca gtacacggtt tccttgaaat
     3481 ttttttgatt cggtaatctc cgaacagaag gaagaacgaa ggaaggagca cagacttaga
     3541 ttggtatata tacgcatatg tagtgttgaa gaaacatgaa attgcccagt attcttaacc
     3601 caactgcaca gaacaaaaac gtgcaggaaa cgaagataaa tcatgtcgaa agctacatat
     3661 aaggaacgtg ctgctactca tcctagtcct gttgctgcca agctatttaa tatcatgcac
     3721 gaaaagcaaa caaacttgtg tgcttcattg gatgttcgta ccaccaagga attactggag
     3781 ttagttgaag cattaggtcc caaaatttgt ttactaaaaa cacatgtgga tatcttgact
     3841 gatttttcca tggagggcac agttaagccg ctaaaggcat tatccgccaa gtacaatttt
     3901 ttactcttcg aggacagaaa atttgctgac attggtaata cagtcaaatt gcagtactct
     3961 gcgggtgtat acagaatagc agaatgggca gacattacga atgcacacgg tgtggtgggc
     4021 ccaggtattg ttagcggttt gaagcaggcg gcagaagaag taacaaagga acctagaggc
     4081 cttttgatgt tagcagaatt gtcatgcaag ggctccctat ctactggaga atatactaag
     4141 ggtactgttg acattgcgaa gagcgacaaa gattttgtta tcggctttat tgctcaaaga
     4201 gacatgggtg gaagagatga aggttacgat tggttgatta tgacacccgg tgtgggttta
     4261 gatgacaagg gagatgcatt gggtcaacag tatagaaccg tggatgatgt ggtttctaca
     4321 ggatctgaca ttattattgt tggaagagga ctatttgcaa agggaaggga tgctaaggta
     4381 gagggtgaac gttacagaaa agcaggctgg gaagcatatt tgagaagatg cggccagcaa
     4441 aactaaaaaa ctgtattata agtaaatgca tgtatactaa actcacaaat tagagcttca
     4501 atttaattat atcagttatt acccgagtaa cgaagcatct gtgcttcatt ttgtagaaca
     4561 aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga
     4621 acagaaatgc aacgcgaaag cgctatttta ccaacgaaga atctgtgctt catttttgta
     4681 aaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt
     4741 acagaacaga aatgcaacgc gagagcgcta ttttaccaac aaagaatcta tacttctttt
     4801 ttgttctaca aaaatgcatc ccgagagcgc tatttttcta acaaagcatc ttagattact
     4861 ttttttctcc tttgtgcgct ctataatgca gtctcttgat aactttttgc actgtaggtc
     4921 cgttaaggtt agaagaaggc tactttggtg tctattttct cttccataaa aaaagcctga
     4981 ctccacttcc cgcgtttact gattactagc gaagctgcgg gtgcattttt tcaagataaa
     5041 ggcatccccg attatattct ataccgatgt ggattgcgca tactttgtga acagaaagtg
     5101 atagcgttga tgattcttca ttggtcagaa aattatgaac ggtttcttct attttgtctc
     5161 tatatactac gtataggaaa tgtttacatt ttcgtattgt tttcgattca ctctatgaat
     5221 agttcttact acaatttttt tgtctaaaga gtaatactag agataaacat aaaaaatgta
     5281 gaggtcgagt ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt tatataggga
     5341 tatagcacag agatatatag caaagagata cttttgagca atgtttgtgg aagcggtatt
     5401 cgcaatattt tagtagctcg ttacagtccg gtgcgttttt ggttttttga aagtgcgtca
     5461 tcagagcgct tttggttttc aaaagcgctc tgaagttcct atactttcta gctagagaat
     5521 aggaacttcc cgagcggccg cgtgttacaa gcttgactcc agcgtaactg gactgcaatc
     5581 aactcactgg ctcaccttca cgggtgggcc tttcttcggt agaaaatcaa aggatcttct
     5641 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     5701 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaggta actggcttca
     5761 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     5821 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5881 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5941 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     6001 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     6061 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     6121 ttccaggggt aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     6181 agcatcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     6241 cagaaaggcc cacccgaagg tgagccaggt gattacattt gggccctcat cagaggtttt
     6301 caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa
     6361 gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg
     6421 ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca
     6481 cttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     6541 tagttgcctg actccccgtc gtgtagataa ctacgatgcg ggagggctta ccatctggcc
     6601 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     6661 accagccagc cgggagggcc gagcgcagaa gtgatcctgc aactttatcc gcctccatcc
     6721 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     6781 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     6841 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     6901 cggttagctc cttcggtcct ccgatcgttg ccagaagtaa gttggccgca gtgttatcac
     6961 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgtg agatgctttt
     7021 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     7081 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     7141 tcatcattgg aaaacgttct tcggggcgta aactctcaag gatcttaccg ctgttgagat
     7201 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     7261 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     7321 cacggaaatg ttgaatactc attttagctt ccttagctcc tgaaaatctc gataactcaa
     7381 aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc
     7441 gatcaagtca aaagcctccg gtcggaggct tttgactttc tgctatggag gtcaggtatg
     7501 atttaaatgg tcagttaga
//
