LOCUS       sequence-300491-        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"
     CDS             complement(390..1193)
                     /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"
     primer_bind     1544..1562
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(2705..2722)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     promoter        complement(5535..5948)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      5994..6025
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        6026..6128
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(6129..6197,6198..6989)
                     /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     complement(6347..6366)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             6990..7181
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      7205..7232
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(7244..7831)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(7853..7882)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      join(7960..8635,1..307)
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 aaatagcgct ctcgggatgc atttttgtag aacaaaaaag aagtatagat tctttgttgg
       61 taaaatagcg ctctcgcgtt gcatttctgt tctgtaaaaa tgcagctcag attctttgtt
      121 tgaaaaatta gcgctctcgc gttgcatttt tgttttacaa aaatgaagca cagattcttc
      181 gttggtaaaa tagcgctttc gcgttgcatt tctgttctgt aaaaatgcag ctcagattct
      241 ttgtttgaaa aattagcgct ctcgcgttgc atttttgttc tacaaaatga agcacagatg
      301 cttcgttact cgggtaataa ctgatataat taaattgaag ctctaatttg tgagtttagt
      361 atacatgcat ttacttataa tacagttttt tagttttgct ggccgcatct tctcaaatat
      421 gcttcccagc ctgcttttct gtaacgttca ccctctacct tagcatccct tccctttgca
      481 aatagtcctc ttccaacaat aataatgtca gatcctgtag aaaccacatc atccacggtt
      541 ctatactgtt gacccaatgc atctcccttg tcatctaaac ccacaccggg tgtcataatc
      601 aaccaatcgt aaccttcatc tcttccaccc atgtctcttt gagcaataaa gccgataaca
      661 aaatctttgt cgctcttcgc aatgtcaaca gtacccttag tatattctcc agtagatagg
      721 gagcccttgc atgacaattc tgctaacatc aaaaggcctc taggttcctt tgttacttct
      781 tctgccgcct gcttcaaacc gctaacaata cctgggccca ccacaccgtg tgcattcgta
      841 atgtctgccc attctgctat tctgtataca cccgcagagt actgcaattt gactgtatta
      901 ccaatgtcag caaattttct gtcctcgaag agtaaaaaat tgtacttggc ggataatgcc
      961 tttagcggct taactgtgcc ctccatggaa aaatcagtca agatatccac atgtgttttt
     1021 agtaaacaaa ttttgggacc taatgcttca actaactcca gtaattcctt ggtggtacga
     1081 acatccaatg aagcacacaa gtttgtttgc ttttcgtgca tgatattaaa tagcttggca
     1141 gcaacaggac taggatgagt agcagcacgt tccttatatg tagctttcga catgatttat
     1201 cttcgtttcc tgcacgtttt tgttctgtgc agttgggtta agaatactgg gcaatttcat
     1261 gtttcttcaa cactacatat gcgtatatat accaatctaa gtctgtgctc cttccttcgt
     1321 tcttccttct gttcggagat taccgaatca aaaaaatttc aaggaaaccg tgtactgcag
     1381 tgcactagtg gttcggcggc cgctttgtac agaaaaaaaa gaaaaatttg aaatataaat
     1441 aacgttctta atactaacat aactattaaa aaaaataaat agggacctag acttcaggtt
     1501 gtctaactcc ttccttttcg gttagagcgg atgtgggagg agggcgtgaa tgtaagcgtg
     1561 acataactaa ttacatgata tcgacaaagg aaaaggggcc tgtttacgtt tgagcatctt
     1621 ttaatgcttc gtcgacggtt tgtctaccgg aagcagcatt aatcacggca gtcctaactg
     1681 cgtaccagaa ggctgacatt tgtgggatgt ttggcataat ttcacccttt tgggcgttct
     1741 ccatggtagc ggcaattctg ggatccttag ccaattcttc ttcgtaagat ttcaaagcga
     1801 cagcacccaa tggcttgtcc ttgttaacgg cctccaaacc ttcgtcagtt aacaagtaat
     1861 tttccaaaaa ttctttagct agctccttgt ttggagaagc agcattgatg ccagcggata
     1921 gaacaccgac aaagggcttg gatggttggc ctttgaatgt aggcaaaaca gtgacaccat
     1981 agtttacttt agaagtatca atgttactcc aggcccatgg accatttatg gtcatagcag
     2041 tttcaccctt gttgaaagcg gcttcggcga tagagtagtc agtgtcagcg ttcatgtgtt
     2101 tattctttat taaatcaacc aagaaggtca aaccagcctt tgcaccagca ttatcaacgc
     2161 cgacatcctt aatgtcatac ttaccgtttt cgtatttaaa tgcgtaacca ccgtcagcag
     2221 cgattaatgg ccaggtgaaa taaggttctt gtaagttaaa catcaaagcg ctttttccct
     2281 tggcctttag ctctttatcc aaagctggaa tttcttccca tgtctttggt gggttaggta
     2341 acaagtcctt attgtagatc aacgataagg cttcaacagc aattggatat gctataagct
     2401 taccgttgta tctgacagca tcccaagtga atgggtacaa tttatcttgg aaggccttgt
     2461 ctggggtgat ctcagccagc aaacctgact gagcatatcc accaaaacga tcgtgggccc
     2521 agaaaataat gtcgggacca tcaccagtag cagcaacttg agggaatttt tcttctaact
     2581 tgtctggatg ttcaaccgtc accttgatac ctgtgtcctt ttcaaatttc ttgcctactt
     2641 ctgccaaacc gttgtaaccc ttatctccat ttatccaaat gactagctta ccttcctcga
     2701 ttttagaacc tctcggcacg agctgcaata ctgtgtgtgg agtgtacatc gcttcgtaaa
     2761 attcgggttc ccagtaacgt gacgtattat cgttcgtcaa catcacagag tacatatcta
     2821 acatgtgacc ggtcaattcg tcatgtaatt ttctaatgta ttgaagatat aaatgaaaaa
     2881 catcagcata ttcttggtta gggtgcttag tcaatgggta agcgtctatg gccagagaga
     2941 caaaccgttc gatcatcaag gtaccgtcgg tcttgactat gtcgtccacg aaacaaccgg
     3001 cacccaaaat tctgcttgga tctgggtagg gtaggtaaac gtagtcgtcc ccttgtttca
     3061 ctaacattgt gtgttgacta caaaattcat gtggaccctt agtcaaatct gtttccgtcc
     3121 agcattttgc ctcggacata aaaacattgt tttggtagta taaaacagac ttgaagtttt
     3181 tgattgaagc gaccaaacct tggctagcgt aagtagaatt gaaacaaaca acagcgtcgt
     3241 ctgacaaaat catcatggag aagtgctttc ttaagtacgc gtaaaattcg ttaacgaagt
     3301 cagtatcgac atctcgattt ctatacaaac attcataaag tctatgttgc aagttgcgca
     3361 catacttatc agcgatcttg ttaccatccg tagacagcaa agcgtttacg ttggcggtaa
     3421 cggcttgaca aatgttaaaa accgagttag cgtaagcagt ggtggcatct ccagacgaag
     3481 ttccgcctgg ctttacatat agactaccac cacacataac catttcagaa aggacttgag
     3541 cacattcgtt ggccaatctg taaaaacggt gggacaatga gcagcatgtg gtatgctttc
     3601 tagccaagac caaggatgcc ataattctta gcatatttgg catggctcta tcacacttag
     3661 ggtagtccca acccataaga tgtgggtttt cgacatcaga gtaaacggtc ttgagcatat
     3721 tgtgccaacc accatagaac tttgaggtac cgatgactac agtagcacct ctagtagcag
     3781 ctatggattt caatagcttc tgatgaaatt gacggttagt catcgtagaa cagatagaaa
     3841 caccagcaac ggttctggcc cgattttttg cagagatagc gtactttaga ttcatttgag
     3901 taattgtagg aataacgttt ctcttggtgt acgcgaacaa agcatcttgg tcctcgtagg
     3961 acatggaatc atagtacaat ctggctttac cccacttgtt gaatgggaag ccagcagact
     4021 tatctaagtt gttgacaatg acttgatttg cattgatgca accaccatca taacagtcaa
     4081 aatatttatc aacaacttcg acgacgaaaa gcaactgtct gatatcacac attgttggta
     4141 agttgtaacg gtaatagtca taatcactga tagcagcatt gccgtcttga gcaaagaaaa
     4201 agtgtttcaa ctcaacacta gaaccttctt tgaaaaaacc cttgctaaca gcaaaatcgt
     4261 agaagtcttt attaaagtta cccggcttta cagtttggaa cgcgacgtta ttagtaagag
     4321 cggcaacgct gaaacaggtt gttctcttat ccagcaataa gtttccagaa gcggcgtgca
     4381 ttgcaggatc agctgcgtag accaacaatt ccttgaagga caatcttgat gagtgcaagt
     4441 tcacgtcttg gttatgaacg acccctaatt ctctgaaatg atatcccgta ctgaccacga
     4501 atggaacacc gtcaacaaaa atctttcgaa ccagcggacc gaaagaggtt ggtggaaaaa
     4561 cagtcgagaa caacacatta aagttagcgc agtgaagaat acatctgtcg tccaaacagt
     4621 tgacacaatt agggtggtag gtttggtccc agtatttaaa atatctgtcg aacaacttta
     4681 aacgttcttc ggtgaaatcg tactttagaa ggtcccattt aatgtatggc ttagttaaat
     4741 cagtgtcaac atgggattcg gcagtcaatg ctctagttag ggtgagtatt ggcattaaca
     4801 gagaatagta agaatcgaca actggtaccc cagaacccgg ggtagtctgg ataaaatcac
     4861 cgaagtcata ccagttaccg ttcaaatctt gattatccaa agtcaaaaca ccaacaatac
     4921 ccgcatttct catagcatcg caaaattgga ctgtctttaa caaagcttgt ctgactcttt
     4981 cacctaagtt agcgtaaacc ctcaagatgt cgggattctc tacgaagtcg taccagtcct
     5041 ttttgttgaa gtaatcatcg tcacagcagt tataggtaac caaaatttcc ttcagagtat
     5101 cacagtttcc ttcgtcaaag tgtcgcaagg cgtaaactaa atcagccata gtgtacttgg
     5161 tcaaccgttg tctgctgata tgtggcacca tgtccccatc aattctgaac ttgaaaaaat
     5221 cgtgtttagc gacggctgga cagtccttga gcaaattgta tattgtttct tcgtgttgat
     5281 agttggaaaa agtatgtctt ttaacaacga agtacgaatc gataagattg tcgtcttcat
     5341 ccttctcctg gaatctacaa cagttggtct tcaaaaattt agcgaaacca gcgaccttgt
     5401 cgttataaat atcaaacgcc ctgtaaacta cgtctgttga agtacccgta ccacaagggg
     5461 ttagacgagc agcactcacg ccgcagactc tatttaagaa agattgtgca tcggcggaca
     5521 ttttgtaatt aaaacttaga ttagattgct atgctttctt tctaatgagc aagaagtaaa
     5581 aaaagttgta atagaacaag aaaaatgaaa ctgaaacttg agaaattgaa gaccgtttat
     5641 taacttaaat atcaatggga ggtcatcgaa agagaaaaaa atcaaaaaaa aaaattttca
     5701 agaaaaagaa acgtgataaa aatttttatt gcctttttcg acgaagaaaa agaaacgagg
     5761 cggtctcttt tttcttttcc aaacctttag tacgggtaat taacgacacc ctagaggaag
     5821 aaagagggga aatttagtat gctgtgcttg ggtgttttga agtggtacgg cgatgcgcgg
     5881 agtccgagaa aatctggaag agtaaaaaag gagtagaaac attttgaagc tatggtgtgt
     5941 ggggcggccg ctctaactga ccatttaaat catacctgac ctccatagca gaaagtcaaa
     6001 agcctccgac cggaggcttt tgacttgatc ggcacgtaag aggttccaac tttcaccata
     6061 atgaaataag atcactaccg ggcgtatttt ttgagttatc gagattttca ggagctaagg
     6121 aagctaaaat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     6181 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     6241 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     6301 tacgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     6361 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     6421 atgacttggt tgagtactca ccagtcacag aaaagcatct cacggatggc atgacagtaa
     6481 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctgg
     6541 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     6601 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     6661 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     6721 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggatcac
     6781 ttctgcgctc ggccctcccg gctggctggt ttattgctga taaatctgga gccggtgagc
     6841 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgcatcgtag
     6901 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     6961 taggtgcctc actgattaag cattggtaag tgaccaaaca ggaaaaaacc gcccttaaca
     7021 tggcccgctt tatcagaagc cagacattaa cgcttctgga gaaactcaac gagctggacg
     7081 cggatgaaca ggcagacatc tgtgaatcgc ttcacgacca cgctgatgag ctttaccgca
     7141 gctgcctcgc gcgtttcggt gatgacggtg aaaacctctg atgagggccc aaatgtaatc
     7201 acctggctca ccttcgggtg ggcctttctg cgttgctggc gtttttccat aggctccgcc
     7261 cccctgacga gcatcacaaa aatcgatgct caagtcagag gtggcgaaac ccgacaggac
     7321 tataaagata ccaggcgttt acccctggaa gctccctcgt gcgctctcct gttccgaccc
     7381 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     7441 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     7501 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     7561 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     7621 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     7681 gaagaacagt atttggtatc tgcgctctgc tgaagccagt tacctcggaa aaagagttgg
     7741 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     7801 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgattttct accgaagaaa
     7861 ggcccacccg tgaaggtgag ccagtgagtt gattgcagtc cagttacgct ggagtcaagc
     7921 ttgtaacacg cggccgctcg ggaagttcct attctctagc tagaaagtat aggaacttca
     7981 gagcgctttt gaaaaccaaa agcgctctga tgacgcactt tcaaaaaacc aaaaacgcac
     8041 cggactgtaa cgagctacta aaatattgcg aataccgctt ccacaaacat tgctcaaaag
     8101 tatctctttg ctatatatct ctgtgctata tccctatata acctacccat ccacctttcg
     8161 ctccttgaac ttgcatctaa actcgacctc tacatttttt atgtttatct ctagtattac
     8221 tctttagaca aaaaaattgt agtaagaact attcatagag tgaatcgaaa acaatacgaa
     8281 aatgtaaaca tttcctatac gtagtatata gagacaaaat agaagaaacc gttcataatt
     8341 ttctgaccaa tgaagaatca tcaacgctat cactttctgt tcacaaagta tgcgcaatcc
     8401 acatcggtat agaatataat cggggatgcc tttatcttga aaaaatgcac ccgcagcttc
     8461 gctagtaatc agtaaacgcg ggaagtggag tcaggctttt tttatggaag agaaaataga
     8521 caccaaagta gccttcttct aaccttaacg gacctacagt gcaaaaagtt atcaagagac
     8581 tgcattatag agcgcacaaa ggagaaaaaa agtaatctaa gatgctttgt tagaa
//
