LOCUS       sequence-291448-        7549 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 7549)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 7549)
  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-m4046531
FEATURES             Location/Qualifiers
     source          1..7549
                     /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             3224..3241
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             3242..3259
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3304..3322)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             3673..4476
                     /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      4559..5541
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      5619..5648
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      5670..6257
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(6269..6296)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(6320..6511)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6512..7303,7304..7372))
                     /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     7135..7154
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7373..7475)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(7476..7507)
                     /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 ttactcgtgc cgagaggttc
     3241 tcaccatcat caccatcatt aaacaggccc cttttccttt gtcgatatca tgtaattagt
     3301 tatgtcacgc ttacattcac gccctcctcc cacatccgct ctaaccgaaa aggaaggagt
     3361 tagacaacct gaagtctagg tccctattta ttttttttaa tagttatgtt agtattaaga
     3421 acgttattta tatttcaaat ttttcttttt tttctgtaca aagcggccgc cgaaccacta
     3481 gtgcactgca gtacacggtt tccttgaaat ttttttgatt cggtaatctc cgaacagaag
     3541 gaagaacgaa ggaaggagca cagacttaga ttggtatata tacgcatatg tagtgttgaa
     3601 gaaacatgaa attgcccagt attcttaacc caactgcaca gaacaaaaac gtgcaggaaa
     3661 cgaagataaa tcatgtcgaa agctacatat aaggaacgtg ctgctactca tcctagtcct
     3721 gttgctgcca agctatttaa tatcatgcac gaaaagcaaa caaacttgtg tgcttcattg
     3781 gatgttcgta ccaccaagga attactggag ttagttgaag cattaggtcc caaaatttgt
     3841 ttactaaaaa cacatgtgga tatcttgact gatttttcca tggagggcac agttaagccg
     3901 ctaaaggcat tatccgccaa gtacaatttt ttactcttcg aggacagaaa atttgctgac
     3961 attggtaata cagtcaaatt gcagtactct gcgggtgtat acagaatagc agaatgggca
     4021 gacattacga atgcacacgg tgtggtgggc ccaggtattg ttagcggttt gaagcaggcg
     4081 gcagaagaag taacaaagga acctagaggc cttttgatgt tagcagaatt gtcatgcaag
     4141 ggctccctat ctactggaga atatactaag ggtactgttg acattgcgaa gagcgacaaa
     4201 gattttgtta tcggctttat tgctcaaaga gacatgggtg gaagagatga aggttacgat
     4261 tggttgatta tgacacccgg tgtgggttta gatgacaagg gagatgcatt gggtcaacag
     4321 tatagaaccg tggatgatgt ggtttctaca ggatctgaca ttattattgt tggaagagga
     4381 ctatttgcaa agggaaggga tgctaaggta gagggtgaac gttacagaaa agcaggctgg
     4441 gaagcatatt tgagaagatg cggccagcaa aactaaaaaa ctgtattata agtaaatgca
     4501 tgtatactaa actcacaaat tagagcttca atttaattat atcagttatt acccgagtaa
     4561 cgaagcatct gtgcttcatt ttgtagaaca aaaatgcaac gcgagagcgc taatttttca
     4621 aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgaaag cgctatttta
     4681 ccaacgaaga atctgtgctt catttttgta aaacaaaaat gcaacgcgag agcgctaatt
     4741 tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc gagagcgcta
     4801 ttttaccaac aaagaatcta tacttctttt ttgttctaca aaaatgcatc ccgagagcgc
     4861 tatttttcta acaaagcatc ttagattact ttttttctcc tttgtgcgct ctataatgca
     4921 gtctcttgat aactttttgc actgtaggtc cgttaaggtt agaagaaggc tactttggtg
     4981 tctattttct cttccataaa aaaagcctga ctccacttcc cgcgtttact gattactagc
     5041 gaagctgcgg gtgcattttt tcaagataaa ggcatccccg attatattct ataccgatgt
     5101 ggattgcgca tactttgtga acagaaagtg atagcgttga tgattcttca ttggtcagaa
     5161 aattatgaac ggtttcttct attttgtctc tatatactac gtataggaaa tgtttacatt
     5221 ttcgtattgt tttcgattca ctctatgaat agttcttact acaatttttt tgtctaaaga
     5281 gtaatactag agataaacat aaaaaatgta gaggtcgagt ttagatgcaa gttcaaggag
     5341 cgaaaggtgg atgggtaggt tatataggga tatagcacag agatatatag caaagagata
     5401 cttttgagca atgtttgtgg aagcggtatt cgcaatattt tagtagctcg ttacagtccg
     5461 gtgcgttttt ggttttttga aagtgcgtca tcagagcgct tttggttttc aaaagcgctc
     5521 tgaagttcct atactttcta gctagagaat aggaacttcc cgagcggccg cgtgttacaa
     5581 gcttgactcc agcgtaactg gactgcaatc aactcactgg ctcaccttca cgggtgggcc
     5641 tttcttcggt agaaaatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5701 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5761 ccaactcttt ttccgaggta actggcttca gcagagcgca gataccaaat actgttcttc
     5821 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     5881 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5941 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     6001 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     6061 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6121 gggtcggaac aggagagcgc acgagggagc ttccaggggt aaacgcctgg tatctttata
     6181 gtcctgtcgg gtttcgccac ctctgacttg agcatcgatt tttgtgatgc tcgtcagggg
     6241 ggcggagcct atggaaaaac gccagcaacg cagaaaggcc cacccgaagg tgagccaggt
     6301 gattacattt gggccctcat cagaggtttt caccgtcatc accgaaacgc gcgaggcagc
     6361 tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg
     6421 cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca
     6481 tgttaagggc ggttttttcc tgtttggtca cttaccaatg cttaatcagt gaggcaccta
     6541 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     6601 ctacgatgcg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     6661 gctcaccggc tccagattta tcagcaataa accagccagc cgggagggcc gagcgcagaa
     6721 gtgatcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     6781 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     6841 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     6901 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     6961 ccagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     7021 ttactgtcat gccatccgtg agatgctttt ctgtgactgg tgagtactca accaagtcat
     7081 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     7141 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgta
     7201 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     7261 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     7321 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc attttagctt
     7381 ccttagctcc tgaaaatctc gataactcaa aaaatacgcc cggtagtgat cttatttcat
     7441 tatggtgaaa gttggaacct cttacgtgcc gatcaagtca aaagcctccg gtcggaggct
     7501 tttgactttc tgctatggag gtcaggtatg atttaaatgg tcagttaga
//
