LOCUS       sequence-297594-        7513 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 7513)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 7513)
  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-m4046641
FEATURES             Location/Qualifiers
     source          1..7513
                     /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(3268..3286)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             3637..4440
                     /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      4523..5505
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      5583..5612
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      5634..6221
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(6233..6260)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(6284..6475)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6476..7267,7268..7336))
                     /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     7099..7118
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7337..7439)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(7440..7471)
                     /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 atggctgatg cacaatcgtt tttaaaccgg gtttgcggtg taagtgcagc
      481 ccgtcttaca ccgtgcggca caggcactag tactgatgtc gtatacaggg cttttgacat
      541 ctacaatgat aaagtagctg gttttgctaa attcctaaaa actaattgtt gtcgcttcca
      601 agaaaaggac gaagatgaca atttaattga ttcttacttt gtagttaaga gacacacttt
      661 ctctaactac caacatgaag aaacaattta taatttactt aaggattgtc cagctgttgc
      721 taaacatgac ttctttaagt ttagaataga cggtgacatg gtaccacata tatcacgtca
      781 acgtcttact aaatacacaa tggcagacct cgtctatgct ttaaggcatt ttgatgaagg
      841 taattgtgac acattaaaag aaatacttgt cacatacaat tgttgtgatg atgattattt
      901 caataaaaag gactggtatg attttgtaga aaacccagat atattacgcg tatacgccaa
      961 cttaggtgaa cgtgtacgcc aagctttgtt aaaaacagta caattctgtg atgccatgcg
     1021 aaatgctggt attgttggtg tactgacatt agataatcaa gatctcaatg gtaactggta
     1081 tgatttcggt gatttcatac aaaccacgcc aggtagtgga gttcctgttg tagattctta
     1141 ttattcattg ttaatgccta tattaacctt gaccagggct ttaactgcag agtcacatgt
     1201 tgacactgac ttaacaaagc cttacattaa gtgggatttg ttaaaatatg acttcacgga
     1261 agagaggtta aaactctttg accgttattt taaatattgg gatcagacat accacccaaa
     1321 ttgtgttaac tgtttggatg acagatgcat tctgcattgt gcaaacttta atgttttatt
     1381 ctctacagtg ttcccaccta caagttttgg accactagtg agaaaaatat ttgttgatgg
     1441 tgttccattt gtagtttcaa ctggatacca cttcagagag ctaggtgttg tacataatca
     1501 ggatgtaaac ttacatagct ctagacttag ttttaaggaa ttacttgtgt atgctgctga
     1561 ccctgctatg cacgctgctt ctggtaatct attactagat aaacgcacta cgtgcttttc
     1621 agtagctgca cttactaaca atgttgcttt tcaaactgtc aaacccggta attttaacaa
     1681 agacttctat gactttgctg tgtctaaggg tttctttaag gaaggaagtt ctgttgaatt
     1741 aaaacacttc ttctttgctc aggatggtaa tgctgctatc agcgattatg actactatcg
     1801 ttataatcta ccaacaatgt gtgatatcag acaactacta tttgtagttg aagttgttga
     1861 taagtacttt gattgttacg atggtggctg tattaatgct aaccaagtca tcgtcaacaa
     1921 cctagacaaa tcagctggtt ttccatttaa taaatggggt aaggctagac tttattatga
     1981 ttcaatgagt tatgaggatc aagatgcact tttcgcatat acaaaacgta atgtcatccc
     2041 tactataact caaatgaatc ttaagtatgc cattagtgca aagaatagag ctcgcaccgt
     2101 agctggtgtc tctatctgta gtactatgac caatagacag tttcatcaaa aattattgaa
     2161 atcaatagcc gccactagag gagctactgt agtaattgga acaagcaaat tctatggtgg
     2221 ttggcacaac atgttaaaaa ctgtttatag tgatgtagaa aaccctcacc ttatgggttg
     2281 ggattatcct aaatgtgata gagccatgcc taacatgctt agaattatgg cctcacttgt
     2341 tcttgctcgc aaacatacaa cgtgttgtag cttgtcacac cgtttctata gattagctaa
     2401 tgagtgtgct caagtattga gtgaaatggt catgtgtggc ggttcactat atgttaaacc
     2461 aggtggaacc tcatcaggag atgccacaac tgcttatgct aatagtgttt ttaacatttg
     2521 tcaagctgtc acggccaatg ttaatgcact tttatctact gatggtaaca aaattgccga
     2581 taagtatgtc cgcaatttac aacacagact ttatgagtgt ctctatagaa atagagatgt
     2641 tgacacagac tttgtgaatg agttttacgc atatttgcgt aaacatttct caatgatgat
     2701 actctctgac gatgctgttg tgtgtttcaa tagcacttat gcatctcaag gtctagtggc
     2761 tagcataaag aactttaagt cagttcttta ttatcaaaac aatgttttta tgtctgaagc
     2821 aaaatgttgg actgagactg accttactaa aggacctcat gaattttgct ctcaacatac
     2881 aatgctagtt aaacagggtg atgattatgt gtaccttcct tacccagatc catcaagaat
     2941 cctaggggcc ggctgttttg tagatgatat cgtaaaaaca gatggtacac ttatgattga
     3001 acggttcgtg tctttagcta tagatgctta cccacttact aaacatccta atcaggagta
     3061 tgctgatgtc tttcatttgt acttacaata cataagaaag ctacatgatg agttaacagg
     3121 acacatgtta gacatgtatt ctgttatgct tactaatgat aacacttcaa ggtattggga
     3181 acctgagttt tatgaggcta tgtacacacc gcatacagtc ttataaacag gccccttttc
     3241 ctttgtcgat atcatgtaat tagttatgtc acgcttacat tcacgccctc ctcccacatc
     3301 cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt
     3361 ttaatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg
     3421 tacaaagcgg ccgccgaacc actagtgcac tgcagtacac ggtttccttg aaattttttt
     3481 gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg agcacagact tagattggta
     3541 tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt aacccaactg
     3601 cacagaacaa aaacgtgcag gaaacgaaga taaatcatgt cgaaagctac atataaggaa
     3661 cgtgctgcta ctcatcctag tcctgttgct gccaagctat ttaatatcat gcacgaaaag
     3721 caaacaaact tgtgtgcttc attggatgtt cgtaccacca aggaattact ggagttagtt
     3781 gaagcattag gtcccaaaat ttgtttacta aaaacacatg tggatatctt gactgatttt
     3841 tccatggagg gcacagttaa gccgctaaag gcattatccg ccaagtacaa ttttttactc
     3901 ttcgaggaca gaaaatttgc tgacattggt aatacagtca aattgcagta ctctgcgggt
     3961 gtatacagaa tagcagaatg ggcagacatt acgaatgcac acggtgtggt gggcccaggt
     4021 attgttagcg gtttgaagca ggcggcagaa gaagtaacaa aggaacctag aggccttttg
     4081 atgttagcag aattgtcatg caagggctcc ctatctactg gagaatatac taagggtact
     4141 gttgacattg cgaagagcga caaagatttt gttatcggct ttattgctca aagagacatg
     4201 ggtggaagag atgaaggtta cgattggttg attatgacac ccggtgtggg tttagatgac
     4261 aagggagatg cattgggtca acagtataga accgtggatg atgtggtttc tacaggatct
     4321 gacattatta ttgttggaag aggactattt gcaaagggaa gggatgctaa ggtagagggt
     4381 gaacgttaca gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa
     4441 aaaactgtat tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa
     4501 ttatatcagt tattacccga gtaacgaagc atctgtgctt cattttgtag aacaaaaatg
     4561 caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa
     4621 atgcaacgcg aaagcgctat tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa
     4681 aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa
     4741 cagaaatgca acgcgagagc gctattttac caacaaagaa tctatacttc ttttttgttc
     4801 tacaaaaatg catcccgaga gcgctatttt tctaacaaag catcttagat tacttttttt
     4861 ctcctttgtg cgctctataa tgcagtctct tgataacttt ttgcactgta ggtccgttaa
     4921 ggttagaaga aggctacttt ggtgtctatt ttctcttcca taaaaaaagc ctgactccac
     4981 ttcccgcgtt tactgattac tagcgaagct gcgggtgcat tttttcaaga taaaggcatc
     5041 cccgattata ttctataccg atgtggattg cgcatacttt gtgaacagaa agtgatagcg
     5101 ttgatgattc ttcattggtc agaaaattat gaacggtttc ttctattttg tctctatata
     5161 ctacgtatag gaaatgttta cattttcgta ttgttttcga ttcactctat gaatagttct
     5221 tactacaatt tttttgtcta aagagtaata ctagagataa acataaaaaa tgtagaggtc
     5281 gagtttagat gcaagttcaa ggagcgaaag gtggatgggt aggttatata gggatatagc
     5341 acagagatat atagcaaaga gatacttttg agcaatgttt gtggaagcgg tattcgcaat
     5401 attttagtag ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc gtcatcagag
     5461 cgcttttggt tttcaaaagc gctctgaagt tcctatactt tctagctaga gaataggaac
     5521 ttcccgagcg gccgcgtgtt acaagcttga ctccagcgta actggactgc aatcaactca
     5581 ctggctcacc ttcacgggtg ggcctttctt cggtagaaaa tcaaaggatc ttcttgagat
     5641 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     5701 gtttgtttgc cggatcaaga gctaccaact ctttttccga ggtaactggc ttcagcagag
     5761 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     5821 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5881 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     5941 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6001 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6061 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6121 gggtaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcatc
     6181 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcagaaa
     6241 ggcccacccg aaggtgagcc aggtgattac atttgggccc tcatcagagg ttttcaccgt
     6301 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt
     6361 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg
     6421 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcacttacc
     6481 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     6541 cctgactccc cgtcgtgtag ataactacga tgcgggaggg cttaccatct ggccccagtg
     6601 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     6661 cagccgggag ggccgagcgc agaagtgatc ctgcaacttt atccgcctcc atccagtcta
     6721 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     6781 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     6841 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     6901 gctccttcgg tcctccgatc gttgccagaa gtaagttggc cgcagtgtta tcactcatgg
     6961 ttatggcagc actgcataat tctcttactg tcatgccatc cgtgagatgc ttttctgtga
     7021 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     7081 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     7141 ttggaaaacg ttcttcgggg cgtaaactct caaggatctt accgctgttg agatccagtt
     7201 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     7261 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     7321 aatgttgaat actcatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata
     7381 cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa
     7441 gtcaaaagcc tccggtcgga ggcttttgac tttctgctat ggaggtcagg tatgatttaa
     7501 atggtcagtt aga
//
