LOCUS       sequence-298053-        8599 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 RNA-dependent RNA 
            polymerase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8599)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 8599)
  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-m4046241
FEATURES             Location/Qualifiers
     source          1..8599
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             3194..3211
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(4354..4372)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             4723..5526
                     /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      5609..6591
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      6669..6698
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      6720..7307
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(7319..7346)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(7370..7561)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(7562..8353,8354..8422))
                     /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     8185..8204
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8423..8525)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(8526..8557)
                     /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 caataaattt cttgaaaatt
      241 tttttttttg atttttttct ctttcgatga cctcccattg atatttaagt taataaacgg
      301 tcttcaattt ctcaagtttc agtttcattt ttcttgttct attacaactt tttttacttc
      361 ttgctcatta gaaagaaagc atagcaatct aatctaagtt ttaattacaa aatggctgat
      421 gcacaatcgt ttttaaaccg ggtttgcggt gtaagtgcag cccgtcttac accgtgcggc
      481 acaggcacta gtactgatgt cgtatacagg gcttttgaca tctacaatga taaagtagct
      541 ggttttgcta aattcctaaa aactaattgt tgtcgcttcc aagaaaagga cgaagatgac
      601 aatttaattg attcttacga gacacacttt ctctaactac caacatgaag aaacaattta
      661 taatttactt aaggattgtc cagctgttgc taaacatgac ttctttaagt ttagaataga
      721 cggtgacatg gtaccacata tatcacgtca acgtcttact aaatacacaa tggcagacct
      781 cgtctatgct ttaaggcatt ttgatgaagg taattgtgac acattaaaag aaatacttgt
      841 cacatacaat tgttgtgatg atgattattt caataaaaag gactggtatg attttgtaga
      901 aaacccagat atattacgcg tatacgccaa cttaggtgaa cgtgtacgcc aagctttgtt
      961 aaaaacagta caattctgtg atgccatgcg aaatgctggt attgttggtg tactgacatt
     1021 agataatcaa gatctcaatg gtaactggta tgatttcggt gatttcatac aaaccacgcc
     1081 aggtagtgga gttcctgttg tagattctta ttattcattg ttaatgccta tattaacctt
     1141 gaccagggct ttaactgcag agtcacatgt tgacactgac ttaacaaagc cttacattaa
     1201 gtgggatttg ttaaaatatg acttcacgga agagaggtta aaactctttg accgttattt
     1261 taaatattgg gatcagacat accacccaaa ttgtgttaac tgtttggatg acagatgcat
     1321 tctgcattgt gcaaacttta atgttttatt ctctacagtg ttcccaccta caagttttgg
     1381 accactagtg agaaaaatat ttgttgatgg tgttccattt gtagtttcaa ctggatacca
     1441 cttcagagag ctaggtgttg tacataatca ggatgtaaac ttacatagct ctagacttag
     1501 ttttaaggaa ttacttgtgt atgctgctga ccctgctatg cacgctgctt ctggtaatct
     1561 attactagat aaacgcacta cgtgcttttc agtagctgca cttactaaca atgttgcttt
     1621 tcaaactgtc aaacccggta attttaacaa agacttctat gactttgctg tgtctaaggg
     1681 tttctttaag gaaggaagtt ctgttgaatt aaaacacttc ttctttgctc aggatggtaa
     1741 tgctgctatc agcgattatg actactatcg ttataatcta ccaacaatgt gtgatatcag
     1801 acaactacta tttgtagttg aagttgttga taagtacttt gattgttacg atggtggctg
     1861 tattaatgct aaccaagtca tcgtcaacaa cctagacaaa tcagctggtt ttccatttaa
     1921 taaatggggt aaggctagac tttattatga ttcaatgagt tatgaggatc aagatgcact
     1981 tttcgcatat acaaaacgta atgtcatccc tactataact caaatgaatc ttaagtatgc
     2041 cattagtgca aagaatagag ctcgcaccgt agctggtgtc tctatctgta gtactatgac
     2101 caatagacag tttcatcaaa aattattgaa atcaatagcc gccactagag gagctactgt
     2161 agtaattgga acaagcaaat tctatggtgg ttggcacaac atgttaaaaa ctgtttatag
     2221 tgatgtagaa aaccctcacc ttatgggttg ggattatcct aaatgtgata gagccatgcc
     2281 taacatgctt agaattatgg cctcacttgt tcttgctcgc aaacatacaa cgtgttgtag
     2341 cttgtcacac cgtttctata gattagctaa tgagtgtgct caagtattga gtgaaatggt
     2401 catgtgtggc ggttcactat atgttaaacc aggtggaacc tcatcaggag atgccacaac
     2461 tgcttatgct aatagtgttt ttaacatttg tcaagctgtc acggccaatg ttaatgcact
     2521 tttatctact gatggtaaca aaattgccga taagtatgtc cgcaatttac aacacagact
     2581 ttatgagtgt ctctatagaa atagagatgt tgacacagac tttgtgaatg agttttacgc
     2641 atatttgcgt aaacatttct caatgatgat actctctgac gatgctgttg tgtgtttcaa
     2701 tagcacttat gcatctcaag gtctagtggc tagcataaag aactttaagt cagttcttta
     2761 ttatcaaaac aatgttttta tgtctgaagc aaaatgttgg actgagactg accttactaa
     2821 aggacctcat gaattttgct ctcaacatac aatgctagtt aaacagggtg atgattatgt
     2881 gtaccttcct tacccagatc catcaagaat cctaggggcc ggctgttttg tagatgatat
     2941 cgtaaaaaca gatggtacac ttatgattga acggttcgtg tctttagcta tagatgctta
     3001 cccacttact aaacatccta atcaggagta tgctgatgtc tttcatttgt acttacaata
     3061 cataagaaag ctacatgatg agttaacagg acacatgtta gacatgtatt ctgttatgct
     3121 tactaatgat aacacttcaa ggtattggga acctgagttt tatgaggcta tgtacacacc
     3181 gcatacagtc ttactcgtgc cgagaggttc taaaatcgag gaaggtaagc tagtcatttg
     3241 gataaatgga gataagggtt acaacggttt ggcagaagta ggcaagaaat ttgaaaagga
     3301 cacaggtatc aaggtgacgg ttgaacatcc agacaagtta gaagaaaaat tccctcaagt
     3361 tgctgctact ggtgatggtc ccgacattat tttctgggcc cacgatcgtt ttggtggata
     3421 tgctcagtca ggtttgctgg ctgagatcac cccagacaag gccttccaag ataaattgta
     3481 cccattcact tgggatgctg tcagatacaa cggtaagctt atagcatatc caattgctgt
     3541 tgaagcctta tcgttgatct acaataagga cttgttacct aacccaccaa agacatggga
     3601 agaaattcca gctttggata aagagctaaa ggccaaggga aaaagcgctt tgatgtttaa
     3661 cttacaagaa ccttatttca cctggccatt aatcgctgct gacggtggtt acgcatttaa
     3721 atacgaaaac ggtaagtatg acattaagga tgtcggcgtt gataatgctg gtgcaaaggc
     3781 tggtttgacc ttcttggttg atttaataaa gaataaacac atgaacgctg acactgacta
     3841 ctctatcgcc gaagccgctt tcaacaaggg tgaaactgct atgaccataa atggtccatg
     3901 ggcctggagt aacattgata cttctaaagt aaactatggt gtcactgttt tgcctacatt
     3961 caaaggccaa ccatccaagc cctttgtcgg tgttctatcc gctggcatca atgctgcttc
     4021 tccaaacaag gagctagcta aagaattttt ggaaaattac ttgttaactg acgaaggttt
     4081 ggaggccgtt aacaaggaca agccattggg tgctgtcgct ttgaaatctt acgaagaaga
     4141 attggctaag gatcccagaa ttgccgctac catggagaac gcccaaaagg gtgaaattat
     4201 gccaaacatc ccacaaatgt cagccttctg gtacgcagtt aggactgccg tgattaatgc
     4261 tgcttccggt agacaaaccg tcgacgaagc attaaaagat gctcaaacgt aaacaggccc
     4321 cttttccttt gtcgatatca tgtaattagt tatgtcacgc ttacattcac gccctcctcc
     4381 cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta
     4441 ttttttttaa tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt
     4501 tttctgtaca aagcggccgc cgaaccacta gtgcactgca gtacacggtt tccttgaaat
     4561 ttttttgatt cggtaatctc cgaacagaag gaagaacgaa ggaaggagca cagacttaga
     4621 ttggtatata tacgcatatg tagtgttgaa gaaacatgaa attgcccagt attcttaacc
     4681 caactgcaca gaacaaaaac gtgcaggaaa cgaagataaa tcatgtcgaa agctacatat
     4741 aaggaacgtg ctgctactca tcctagtcct gttgctgcca agctatttaa tatcatgcac
     4801 gaaaagcaaa caaacttgtg tgcttcattg gatgttcgta ccaccaagga attactggag
     4861 ttagttgaag cattaggtcc caaaatttgt ttactaaaaa cacatgtgga tatcttgact
     4921 gatttttcca tggagggcac agttaagccg ctaaaggcat tatcngccaa gtacaatttt
     4981 ttactcttcg aggacagaaa atttgctgac attggtaata cagtcaaatt gcagtactct
     5041 gcgggtgtat acagaatagc agaatgggca gacattacga atgcacacgg tgtggtgggc
     5101 ccaggtattg ttagcggttt gaagcaggcg gcagaagaag taacaaagga acctagaggc
     5161 cttttgatgt tagcagaatt gtcatgcaag ggctccctat ctactggaga atatactaag
     5221 ggtactgttg acattgcgaa gagcgacaaa gattttgtta tcggctttat tgctcaaaga
     5281 gacatgggtg gaagagatga aggttacgat tggttgatta tgacacccgg tgtgggttta
     5341 gatgacaagg gagatgcatt gggtcaacag tatagaaccg tggatgatgt ggtttctaca
     5401 ggatctgaca ttattattgt tggaagagga ctatttgcaa agggaaggga tgctaaggta
     5461 gagggtgaac gttacagaaa agcaggctgg gaagcatatt tgagaagatg cggccagcaa
     5521 aactaaaaaa ctgtattata agtaaatgca tgtatactaa actcacaaat tagagcttca
     5581 atttaattat atcagttatt acccgagtaa cgaagcatct gtgcttcatt ttgtagaaca
     5641 aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga
     5701 acagaaatgc aacgcgaaag cgctatttta ccaacgaaga atctgtgctt catttttgta
     5761 aaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt
     5821 acagaacaga aatgcaacgc gagagcgcta ttttaccaac aaagaatcta tacttctttt
     5881 ttgttctaca aaaatgcatc ccgagagcgc tattttycta acaaagcatc ttagattact
     5941 ttttttctcc tttgtgcgct ctataatgca gtctcttgat aactttttgc actgtaggtc
     6001 cgttaaggtt agaagaaggc tactttggtg tctattttct cttccataaa aaaagcctga
     6061 ctccacttcc cgcgtttact gattactagc gaagctgcgg gtgcattttt tcaagataaa
     6121 ggcatccccg attatattct ataccgatgt ggattgcgca tactttgtga acagaaagtg
     6181 atagcgttga tgattcttca ttggtcagaa aattatgaac ggtttcttct attttgtctc
     6241 tatatactac gtataggaaa tgtttacatt ttcgtattgt tttcgattca ctctatgaat
     6301 agttcttact acaatttttt tgtctaaaga gtaatactag agataaacat aaaaaatgta
     6361 gaggtcgagt ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt tatataggga
     6421 tatagcacag agatatatag caaagagata cttttgagca atgtttgtgg aagcggtatt
     6481 cgcaatattt tagtagctcg ttacagtccg gtgcgttttt ggttttttga aagtgcgtca
     6541 tcagagcgct tttggttttc aaaagcgctc tgaagttcct atactttcta gctagagaat
     6601 aggaacttcc cgagcggccg cgtgttacaa gcttgactcc agcgtaactg gactgcaatc
     6661 aactcactgg ctcaccttca cgggtgggcc tttcttcggt agaaaatcaa aggatcttct
     6721 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     6781 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaggta actggcttca
     6841 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     6901 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     6961 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     7021 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     7081 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     7141 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     7201 ttccaggggt aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     7261 agcatcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     7321 cagaaaggcc cacccgaagg tgagccaggt gattacattt gggccctcat cagaggtttt
     7381 caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa
     7441 gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg
     7501 ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca
     7561 cttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     7621 tagttgcctg actccccgtc gtgtagataa ctacgatgcg ggagggctta ccatctggcc
     7681 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     7741 accagccagc cgggagggcc gagcgcagaa gtgatcctgc aactttatcc gcctccatcc
     7801 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     7861 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     7921 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     7981 cggttagctc cttcggtcct ccgatcgttg ccagaagtaa gttggccgca gtgttatcac
     8041 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgtg agatgctttt
     8101 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     8161 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     8221 tcatcattgg aaaacgttct tcggggcgta aactctcaag gatcttaccg ctgttgagat
     8281 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     8341 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     8401 cacggaaatg ttgaatactc attttagctt ccttagctcc tgaaaatctc gataactcaa
     8461 aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc
     8521 gatcaagtca aaagcctccg gtcggaggct tttgactttc tgctatggag gtcaggtatg
     8581 atttaaatgg tcagttaga
//
