LOCUS       sequence-297615-        8635 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 8635)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 8635)
  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-m4046219
FEATURES             Location/Qualifiers
     source          1..8635
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             3230..3247
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(4390..4408)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             4759..5562
                     /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      5645..6627
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      6705..6734
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      6756..7343
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(7355..7382)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(7406..7597)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(7598..8389,8390..8458))
                     /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     8221..8240
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8459..8561)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(8562..8593)
                     /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 gtattgcagc tcgtgccgag
     3241 aggttctaaa atcgaggaag gtaagctagt catttggata aatggagata agggttacaa
     3301 cggtttggca gaagtaggca agaaatttga aaaggacaca ggtatcaagg tgacggttga
     3361 acatccagac aagttagaag aaaaattccc tcaagttgct gctactggtg atggtcccga
     3421 cattattttc tgggcccacg atcgttttgg tggatatgct cagtcaggtt tgctggctga
     3481 gatcacccca gacaaggcct tccaagataa attgtaccca ttcacttggg atgctgtcag
     3541 atacaacggt aagcttatag catatccaat tgctgttgaa gccttatcgt tgatctacaa
     3601 taaggacttg ttacctaacc caccaaagac atgggaagaa attccagctt tggataaaga
     3661 gctaaaggcc aagggaaaaa gcgctttgat gtttaactta caagaacctt atttcacctg
     3721 gccattaatc gctgctgacg gtggttacgc atttaaatac gaaaacggta agtatgacat
     3781 taaggatgtc ggcgttgata atgctggtgc aaaggctggt ttgaccttct tggttgattt
     3841 aataaagaat aaacacatga acgctgacac tgactactct atcgccgaag ccgctttcaa
     3901 caagggtgaa actgctatga ccataaatgg tccatgggcc tggagtaaca ttgatacttc
     3961 taaagtaaac tatggtgtca ctgttttgcc tacattcaaa ggccaaccat ccaagccctt
     4021 tgtcggtgtt ctatccgctg gcatcaatgc tgcttctcca aacaaggagc tagctaaaga
     4081 atttttggaa aattacttgt taactgacga aggtttggag gccgttaaca aggacaagcc
     4141 attgggtgct gtcgctttga aatcttacga agaagaattg gctaaggatc ccagaattgc
     4201 cgctaccatg gagaacgccc aaaagggtga aattatgcca aacatcccac aaatgtcagc
     4261 cttctggtac gcagttagga ctgccgtgat taatgctgct tccggtagac aaaccgtcga
     4321 cgaagcatta aaagatgctc aaacgtaaac aggccccttt tcctttgtcg atatcatgta
     4381 attagttatg tcacgcttac attcacgccc tcctcccaca tccgctctaa ccgaaaagga
     4441 aggagttaga caacctgaag tctaggtccc tatttatttt ttttaatagt tatgttagta
     4501 ttaagaacgt tatttatatt tcaaattttt cttttttttc tgtacaaagc ggccgccgaa
     4561 ccactagtgc actgcagtac acggtttcct tgaaattttt ttgattcggt aatctccgaa
     4621 cagaaggaag aacgaaggaa ggagcacaga cttagattgg tatatatacg catatgtagt
     4681 gttgaagaaa catgaaattg cccagtattc ttaacccaac tgcacagaac aaaaacgtgc
     4741 aggaaacgaa gataaatcat gtcgaaagct acatataagg aacgtgctgc tactcatcct
     4801 agtcctgttg ctgccaagct atttaatatc atgcacgaaa agcaaacaaa cttgtgtgct
     4861 tcattggatg ttcgtaccac caaggaatta ctggagttag ttgaagcatt aggtcccaaa
     4921 atttgtttac taaaaacaca tgtggatatc ttgactgatt tttccatgga gggcacagtt
     4981 aagccgctaa aggcattatc cgccaagtac aattttttac tcttcgagga cagaaaattt
     5041 gctgacattg gtaatacagt caaattgcag tactctgcgg gtgtatacag aatagcagaa
     5101 tgggcagaca ttacgaatgc acacggtgtg gtgggcccag gtattgttag cggtttgaag
     5161 caggcggcag aagaagtaac aaaggaacct agaggccttt tgatgttagc agaattgtca
     5221 tgcaagggct ccctatctac tggagaatat actaagggta ctgttgacat tgcgaagagc
     5281 gacaaagatt ttgttatcgg ctttattgct caaagagaca tgggtggaag agatgaaggt
     5341 tacgattggt tgattatgac acccggtgtg ggtttagatg acaagggaga tgcattgggt
     5401 caacagtata gaaccgtgga tgatgtggtt tctacaggat ctgacattat tattgttgga
     5461 agaggactat ttgcaaaggg aagggatgct aaggtagagg gtgaacgtta cagaaaagca
     5521 ggctgggaag catatttgag aagatgcggc cagcaaaact aaaaaactgt attataagta
     5581 aatgcatgta tactaaactc acaaattaga gcttcaattt aattatatca gttattaccc
     5641 gagtaacgaa gcatctgtgc ttcattttgt agaacaaaaa tgcaacgcga gagcgctaat
     5701 ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg cgaaagcgct
     5761 attttaccaa cgaagaatct gtgcttcatt tttgtaaaac aaaaatgcaa cgcgagagcg
     5821 ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg caacgcgaga
     5881 gcgctatttt accaacaaag aatctatact tcttttttgt tctacaaaaa tgcatcccga
     5941 gagcgctatt tttctaacaa agcatcttag attacttttt ttctcctttg tgcgctctat
     6001 aatgcagtct cttgataact ttttgcactg taggtccgtt aaggttagaa gaaggctact
     6061 ttggtgtcta ttttctcttc cataaaaaaa gcctgactcc acttcccgcg tttactgatt
     6121 actagcgaag ctgcgggtgc attttttcaa gataaaggca tccccgatta tattctatac
     6181 cgatgtggat tgcgcatact ttgtgaacag aaagtgatag cgttgatgat tcttcattgg
     6241 tcagaaaatt atgaacggtt tcttctattt tgtctctata tactacgtat aggaaatgtt
     6301 tacattttcg tattgttttc gattcactct atgaatagtt cttactacaa tttttttgtc
     6361 taaagagtaa tactagagat aaacataaaa aatgtagagg tcgagtttag atgcaagttc
     6421 aaggagcgaa aggtggatgg gtaggttata tagggatata gcacagagat atatagcaaa
     6481 gagatacttt tgagcaatgt ttgtggaagc ggtattcgca atattttagt agctcgttac
     6541 agtccggtgc gtttttggtt ttttgaaagt gcgtcatcag agcgcttttg gttttcaaaa
     6601 gcgctctgaa gttcctatac tttctagcta gagaatagga acttcccgag cggccgcgtg
     6661 ttacaagctt gactccagcg taactggact gcaatcaact cactggctca ccttcacggg
     6721 tgggcctttc ttcggtagaa aatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6781 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6841 gagctaccaa ctctttttcc gaggtaactg gcttcagcag agcgcagata ccaaatactg
     6901 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6961 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     7021 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     7081 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     7141 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     7201 gcggcagggt cggaacagga gagcgcacga gggagcttcc aggggtaaac gcctggtatc
     7261 tttatagtcc tgtcgggttt cgccacctct gacttgagca tcgatttttg tgatgctcgt
     7321 caggggggcg gagcctatgg aaaaacgcca gcaacgcaga aaggcccacc cgaaggtgag
     7381 ccaggtgatt acatttgggc cctcatcaga ggttttcacc gtcatcaccg aaacgcgcga
     7441 ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt
     7501 catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc
     7561 gggccatgtt aagggcggtt ttttcctgtt tggtcactta ccaatgctta atcagtgagg
     7621 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     7681 agataactac gatgcgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     7741 acccacgctc accggctcca gatttatcag caataaacca gccagccggg agggccgagc
     7801 gcagaagtga tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     7861 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     7921 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     7981 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     8041 tcgttgccag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     8101 attctcttac tgtcatgcca tccgtgagat gcttttctgt gactggtgag tactcaacca
     8161 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     8221 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     8281 ggcgtaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     8341 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     8401 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcattt
     8461 tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta
     8521 tttcattatg gtgaaagttg gaacctctta cgtgccgatc aagtcaaaag cctccggtcg
     8581 gaggcttttg actttctgct atggaggtca ggtatgattt aaatggtcag ttaga
//
