LOCUS       sequence-307320-        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"
     primer_bind     complement(2857..2875)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             3226..4029
                     /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      4112..5094
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      5172..5201
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      5223..5810
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5822..5849)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5873..6064)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6065..6856,6857..6925))
                     /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     6688..6707
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6926..7028)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(7029..7060)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        7106..7519
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
ORIGIN
        1 ttttaattac aaaatgtccg ccgatgcaca atctttctta aatagagtct gcggcgtgag
       61 tgctgctcgt ctaacccctt gtggtacggg tacttcaaca gacgtagttt acagggcgtt
      121 tgatatttat aacgacaagg tcgctggttt cgctaaattt ttgaagacca actgttgtag
      181 attccaggag aaggatgaag acgacaatct tatcgattcg tacttcgttg ttaaaagaca
      241 tactttttcc aactatcaac acgaagaaac aatatacaat ttgctcaagg actgtccagc
      301 cgtcgctaaa cacgattttt tcaagttcag aattgatggg gacatggtgc cacatatcag
      361 cagacaacgg ttgaccaagt acactatggc tgatttagtt tacgccttgc gacactttga
      421 cgaaggaaac tgtgatactc tgaaggaaat tttggttacc tataactgct gtgacgatga
      481 ttacttcaac aaaaaggact ggtacgactt cgtagagaat cccgacatct tgagggttta
      541 cgctaactta ggtgaaagag tcagacaagc tttgttaaag acagtccaat tttgcgatgc
      601 tatgagaaat gcgggtattg ttggtgtttt gactttggat aatcaagatt tgaacggtaa
      661 ctggtatgac ttcggtgatt ttatccagac taccccgggt tctggggtac cagttgtcga
      721 ttcttactat tctctgttaa tgccaatact caccctaact agagcattga ctgccgaatc
      781 ccatgttgac actgatttaa ctaagccata cattaaatgg gaccttctaa agtacgattt
      841 caccgaagaa cgtttaaagt tgttcgacag atattttaaa tactgggacc aaacctacca
      901 ccctaattgt gtcaactgtt tggacgacag atgtattctt cactgcgcta actttaatgt
      961 gttgttctcg actgtttttc caccaacctc tttcggtccg ctggttcgaa agatttttgt
     1021 tgacggtgtt ccattcgtgg tcagtacggg atatcatttc agagaattag gggtcgttca
     1081 taaccaagac gtgaacttgc actcatcaag attgtccttc aaggaattgt tggtctacgc
     1141 agctgatcct gcaatgcacg ccgcttctgg aaacttattg ctggataaga gaacaacctg
     1201 tttcagcgtt gccgctctta ctaataacgt cgcgttccaa actgtaaagc cgggtaactt
     1261 taataaagac ttctacgatt ttgctgttag caagggtttt ttcaaagaag gttctagtgt
     1321 tgagttgaaa cactttttct ttgctcaaga cggcaatgct gctatcagtg attatgacta
     1381 ttaccgttac aacttaccaa caatgtgtga tatcagacag ttgcttttcg tcgtcgaagt
     1441 tgttgataaa tattttgact gttatgatgg tggttgcatc aatgcaaatc aagtcattgt
     1501 caacaactta gataagtctg ctggcttccc attcaacaag tggggtaaag ccagattgta
     1561 ctatgattcc atgtcctacg aggaccaaga tgctttgttc gcgtacacca agagaaacgt
     1621 tattcctaca attactcaaa tgaatctaaa gtacgctatc tctgcaaaaa atcgggccag
     1681 aaccgttgct ggtgtttcta tctgttctac gatgactaac cgtcaatttc atcagaagct
     1741 attgaaatcc atagctgcta ctagaggtgc tactgtagtc atcggtacct caaagttcta
     1801 tggtggttgg cacaatatgc tcaagaccgt ttactctgat gtcgaaaacc cacatcttat
     1861 gggttgggac taccctaagt gtgatagagc catgccaaat atgctaagaa ttatggcatc
     1921 cttggtcttg gctagaaagc ataccacatg ctgctcattg tcccaccgtt tttacagatt
     1981 ggccaacgaa tgtgctcaag tcctttctga aatggttatg tgtggtggta gtctatatgt
     2041 aaagccaggc ggaacttcgt ctggagatgc caccactgct tacgctaact cggtttttaa
     2101 catttgtcaa gccgttaccg ccaacgtaaa cgctttgctg tctacggatg gtaacaagat
     2161 cgctgataag tatgtgcgca acttgcaaca tagactttat gaatgtttgt atagaaatcg
     2221 agatgtcgat actgacttcg ttaacgaatt ttacgcgtac ttaagaaagc acttctccat
     2281 gatgattttg tcagacgacg ctgttgtttg tttcaattct acttacgcta gccaaggttt
     2341 ggtcgcttca atcaaaaact tcaagtctgt tttatactac caaaacaatg tttttatgtc
     2401 cgaggcaaaa tgctggacgg aaacagattt gactaagggt ccacatgaat tttgtagtca
     2461 acacacaatg ttagtgaaac aaggggacga ctacgtttac ctaccctacc cagatccaag
     2521 cagaattttg ggtgccggtt gtttcgtgga cgacatagtc aagaccgacg gtaccttgat
     2581 gatcgaacgg tttgtctctc tggccataga cgcttaccca ttgactaagc accctaacca
     2641 agaatatgct gatgtttttc atttatatct tcaatacatt agaaaattac atgacgaatt
     2701 gaccggtcac atgttagata tgtactctgt gatgttgacg aacgataata cgtcacgtta
     2761 ctgggaaccc gaattttacg aagcgatgta cactccacac acagtattgc agtaaacagg
     2821 ccccttttcc tttgtcgata tcatgtaatt agttatgtca cgcttacatt cacgccctcc
     2881 tcccacatcc gctctaaccg aaaaggaagg agttagacaa cctgaagtct aggtccctat
     2941 ttattttttt taatagttat gttagtatta agaacgttat ttatatttca aatttttctt
     3001 ttttttctgt acaaagcggc cgccgaacca ctagtgcact gcagtacacg gtttccttga
     3061 aatttttttg attcggtaat ctccgaacag aaggaagaac gaaggaagga gcacagactt
     3121 agattggtat atatacgcat atgtagtgtt gaagaaacat gaaattgccc agtattctta
     3181 acccaactgc acagaacaaa aacgtgcagg aaacgaagat aaatcatgtc gaaagctaca
     3241 tataaggaac gtgctgctac tcatcctagt cctgttgctg ccaagctatt taatatcatg
     3301 cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc gtaccaccaa ggaattactg
     3361 gagttagttg aagcattagg tcccaaaatt tgtttactaa aaacacatgt ggatatcttg
     3421 actgattttt ccatggaggg cacagttaag ccgctaaagg cattatccgc caagtacaat
     3481 tttttactct tcgaggacag aaaatttgct gacattggta atacagtcaa attgcagtac
     3541 tctgcgggtg tatacagaat agcagaatgg gcagacatta cgaatgcaca cggtgtggtg
     3601 ggcccaggta ttgttagcgg tttgaagcag gcggcagaag aagtaacaaa ggaacctaga
     3661 ggccttttga tgttagcaga attgtcatgc aagggctccc tatctactgg agaatatact
     3721 aagggtactg ttgacattgc gaagagcgac aaagattttg ttatcggctt tattgctcaa
     3781 agagacatgg gtggaagaga tgaaggttac gattggttga ttatgacacc cggtgtgggt
     3841 ttagatgaca agggagatgc attgggtcaa cagtatagaa ccgtggatga tgtggtttct
     3901 acaggatctg acattattat tgttggaaga ggactatttg caaagggaag ggatgctaag
     3961 gtagagggtg aacgttacag aaaagcaggc tgggaagcat atttgagaag atgcggccag
     4021 caaaactaaa aaactgtatt ataagtaaat gcatgtatac taaactcaca aattagagct
     4081 tcaatttaat tatatcagtt attacccgag taacgaagca tctgtgcttc attttgtaga
     4141 acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac
     4201 agaacagaaa tgcaacgcga aagcgctatt ttaccaacga agaatctgtg cttcattttt
     4261 gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt
     4321 tttacagaac agaaatgcaa cgcgagagcg ctattttacc aacaaagaat ctatacttct
     4381 tttttgttct acaaaaatgc atcccgagag cgctattttt ctaacaaagc atcttagatt
     4441 actttttttc tcctttgtgc gctctataat gcagtctctt gataactttt tgcactgtag
     4501 gtccgttaag gttagaagaa ggctactttg gtgtctattt tctcttccat aaaaaaagcc
     4561 tgactccact tcccgcgttt actgattact agcgaagctg cgggtgcatt ttttcaagat
     4621 aaaggcatcc ccgattatat tctataccga tgtggattgc gcatactttg tgaacagaaa
     4681 gtgatagcgt tgatgattct tcattggtca gaaaattatg aacggtttct tctattttgt
     4741 ctctatatac tacgtatagg aaatgtttac attttcgtat tgttttcgat tcactctatg
     4801 aatagttctt actacaattt ttttgtctaa agagtaatac tagagataaa cataaaaaat
     4861 gtagaggtcg agtttagatg caagttcaag gagcgaaagg tggatgggta ggttatatag
     4921 ggatatagca cagagatata tagcaaagag atacttttga gcaatgtttg tggaagcggt
     4981 attcgcaata ttttagtagc tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg
     5041 tcatcagagc gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt ctagctagag
     5101 aataggaact tcccgagcgg ccgcgtgtta caagcttgac tccagcgtaa ctggactgca
     5161 atcaactcac tggctcacct tcacgggtgg gcctttcttc ggtagaaaat caaaggatct
     5221 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     5281 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgag gtaactggct
     5341 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     5401 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     5461 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     5521 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     5581 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     5641 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     5701 agcttccagg ggtaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     5761 ttgagcatcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     5821 acgcagaaag gcccacccga aggtgagcca ggtgattaca tttgggccct catcagaggt
     5881 tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt
     5941 gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa
     6001 gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg
     6061 tcacttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     6121 ccatagttgc ctgactcccc gtcgtgtaga taactacgat gcgggagggc ttaccatctg
     6181 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     6241 taaaccagcc agccgggagg gccgagcgca gaagtgatcc tgcaacttta tccgcctcca
     6301 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6361 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6421 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     6481 aagcggttag ctccttcggt cctccgatcg ttgccagaag taagttggcc gcagtgttat
     6541 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtgagatgct
     6601 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6661 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     6721 tgctcatcat tggaaaacgt tcttcggggc gtaaactctc aaggatctta ccgctgttga
     6781 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     6841 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     6901 cgacacggaa atgttgaata ctcattttag cttccttagc tcctgaaaat ctcgataact
     6961 caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt
     7021 gccgatcaag tcaaaagcct ccggtcggag gcttttgact ttctgctatg gaggtcaggt
     7081 atgatttaaa tggtcagtta gagcggccgc cccacacacc atagcttcaa aatgtttcta
     7141 ctcctttttt actcttccag attttctcgg actccgcgca tcgccgtacc acttcaaaac
     7201 acccaagcac agcatactaa atttcccctc tttcttcctc tagggtgtcg ttaattaccc
     7261 gtactaaagg tttggaaaag aaaaaagaga ccgcctcgtt tctttttctt cgtcgaaaaa
     7321 ggcaataaaa atttttatca cgtttctttt tcttgaaaat tttttttttt gatttttttc
     7381 tctttcgatg acctcccatt gatatttaag ttaataaacg gtcttcaatt tctcaagttt
     7441 cagtttcatt tttcttgttc tattacaact ttttttactt cttgctcatt agaaagaaag
     7501 catagcaatc taatctaag
//
