LOCUS       sequence-287722-        5892 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 endoRNAse.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5892)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5892)
  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-m4046367
FEATURES             Location/Qualifiers
     source          1..5892
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        25..466
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    25..142
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     437..460
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             1519..1536
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1537..1602
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(1647..1665)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2016..2819
                     /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      2902..3884
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3962..3991
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4013..4600
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4612..4639)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4663..4854)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4855..5646,5647..5715))
                     /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     5478..5497
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5716..5818)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5819..5850)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgctt gctatattga agtacggatt agaagccgcc gagcgggcga cagccctccg
       61 acggaagact ctcctccgtg cgtcctcgtc ttcaccggtc gcgttcctga aacgcagatg
      121 tgcctcgcgc cgcactgctc cgaacaataa agattctaca atactagctt ttatggttat
      181 gaagaggaaa aattggcagt aacctggccc cacaaacctt caaattaacg aatcaaatta
      241 acaaccatag gatgataatg cgattagttt tttagcctta tttctggggt aattaatcag
      301 cgaagcgatg atttttgatc tattaacaga tatataaatg gaaaagctgc ataaccactt
      361 taactaatac tttcaacatt ttcagtttgt attacttctt attcaaatgt cataaaagta
      421 tcaacaaaaa attgttaata tacctctata ctttaacgtc aaggagaaaa aactataatg
      481 agcctggaaa acgttgcctt caatgtagtg aacaaaggac actttgatgg gcaacagggt
      541 gaggttcctg tctccattat aaataacacc gtttatacta aggtcgacgg cgttgatgtg
      601 gaattgttcg aaaataagac gacattaccc gtcaacgttg catttgaact ttgggcgaag
      661 cgtaacatca aaccagtccc agaagttaag attttgaaca atttgggtgt tgacatcgct
      721 gctaataccg tcatttggga ttacaagaga gatgctccgg cccatatctc aactataggt
      781 gtctgctcga tgactgacat tgctaaaaag ccaaccgaga ctatctgtgc tccattaacc
      841 gttttctttg acggtagagt agatggtcaa gtcgacctct tcagaaacgc aaggaacggt
      901 gttttgatta cagaaggtag tgttaaaggt ctacaacctt ctgtaggtcc aaagcaagcg
      961 tccttgaacg gcgtcacgtt gattggcgaa gccgttaaga ctcaattcaa ttactataag
     1021 aaggtcgatg gtgttgtgca acaattacca gaaacttact tcactcagtc tcgcaatttg
     1081 caagaattta aaccacgatc tcaaatggaa atcgattttc ttgaactagc tatggacgag
     1141 ttcatcgaac gttacaagtt ggaaggttat gcttttgaac acattgtcta cggtgacttc
     1201 tctcattccc agctgggtgg tttacacttg ctaattggac tggctaaaag attcaaggaa
     1261 tcaccattcg agttggaaga ttttatccct atggactcta ccgttaaaaa ctacttcatc
     1321 actgatgctc aaaccggttc gagtaagtgt gtttgttccg tcattgactt gttactagat
     1381 gatttcgtag aaataattaa gtctcaagac ttgtcagttg tttccaaggt tgtcaaggta
     1441 accattgact atacggaaat ctcatttatg ttgtggtgca aagacggtca tgttgaaacc
     1501 ttctacccga aattgcaact cgtgccgaga ggttctgatt ataaagatca tgatggtgat
     1561 tataaagatc atgatattga ttataaagat gatgatgata aataaacagg ccccttttcc
     1621 tttgtcgata tcatgtaatt agttatgtca cgcttacatt cacgccctcc tcccacatcc
     1681 gctctaaccg aaaaggaagg agttagacaa cctgaagtct aggtccctat ttattttttt
     1741 taatagttat gttagtatta agaacgttat ttatatttca aatttttctt ttttttctgt
     1801 acaaagcggc cgccgaacca ctagtgcact gcagtacacg gtttccttga aatttttttg
     1861 attcggtaat ctccgaacag aaggaagaac gaaggaagga gcacagactt agattggtat
     1921 atatacgcat atgtagtgtt gaagaaacat gaaattgccc agtattctta acccaactgc
     1981 acagaacaaa aacgtgcagg aaacgaagat aaatcatgtc gaaagctaca tataaggaac
     2041 gtgctgctac tcatcctagt cctgttgctg ccaagctatt taatatcatg cacgaaaagc
     2101 aaacaaactt gtgtgcttca ttggatgttc gtaccaccaa ggaattactg gagttagttg
     2161 aagcattagg tcccaaaatt tgtttactaa aaacacatgt ggatatcttg actgattttt
     2221 ccatggaggg cacagttaag ccgctaaagg cattatccgc caagtacaat tttttactct
     2281 tcgaggacag aaaatttgct gacattggta atacagtcaa attgcagtac tctgcgggtg
     2341 tatacagaat agcagaatgg gcagacatta cgaatgcaca cggtgtggtg ggcccaggta
     2401 ttgttagcgg tttgaagcag gcggcagaag aagtaacaaa ggaacctaga ggccttttga
     2461 tgttagcaga attgtcatgc aagggctccc tatctactgg agaatatact aagggtactg
     2521 ttgacattgc gaagagcgac aaagattttg ttatcggctt tattgctcaa agagacatgg
     2581 gtggaagaga tgaaggttac gattggttga ttatgacacc cggtgtgggt ttagatgaca
     2641 agggagatgc attgggtcaa cagtatagaa ccgtggatga tgtggtttct acaggatctg
     2701 acattattat tgttggaaga ggactatttg caaagggaag ggatgctaag gtagagggtg
     2761 aacgttacag aaaagcaggc tgggaagcat atttgagaag atgcggccag caaaactaaa
     2821 aaactgtatt ataagtaaat gcatgtatac taaactcaca aattagagct tcaatttaat
     2881 tatatcagtt attacccgag taacgaagca tctgtgcttc attttgtaga acaaaaatgc
     2941 aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa
     3001 tgcaacgcga aagcgctatt ttaccaacga agaatctgtg cttcattttt gtaaaacaaa
     3061 aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac
     3121 agaaatgcaa cgcgagagcg ctattttacc aacaaagaat ctatacttct tttttgttct
     3181 acaaaaatgc atcccgagag cgctattttt ctaacaaagc atcttagatt actttttttc
     3241 tcctttgtgc gctctataat gcagtctctt gataactttt tgcactgtag gtccgttaag
     3301 gttagaagaa ggctactttg gtgtctattt tctcttccat aaaaaaagcc tgactccact
     3361 tcccgcgttt actgattact agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc
     3421 ccgattatat tctataccga tgtggattgc gcatactttg tgaacagaaa gtgatagcgt
     3481 tgatgattct tcattggtca gaaaattatg aacggtttct tctattttgt ctctatatac
     3541 tacgtatagg aaatgtttac attttcgtat tgttttcgat tcactctatg aatagttctt
     3601 actacaattt ttttgtctaa agagtaatac tagagataaa cataaaaaat gtagaggtcg
     3661 agtttagatg caagttcaag gagcgaaagg tggatgggta ggttatatag ggatatagca
     3721 cagagatata tagcaaagag atacttttga gcaatgtttg tggaagcggt attcgcaata
     3781 ttttagtagc tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg tcatcagagc
     3841 gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt ctagctagag aataggaact
     3901 tcccgagcgg ccgcgtgtta caagcttgac tccagcgtaa ctggactgca atcaactcac
     3961 tggctcacct tcacgggtgg gcctttcttc ggtagaaaat caaaggatct tcttgagatc
     4021 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     4081 tttgtttgcc ggatcaagag ctaccaactc tttttccgag gtaactggct tcagcagagc
     4141 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     4201 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     4261 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4321 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4381 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4441 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4501 ggtaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcatcg
     4561 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcagaaag
     4621 gcccacccga aggtgagcca ggtgattaca tttgggccct catcagaggt tttcaccgtc
     4681 atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc
     4741 acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt
     4801 ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcacttacca
     4861 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4921 ctgactcccc gtcgtgtaga taactacgat gcgggagggc ttaccatctg gccccagtgc
     4981 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     5041 agccgggagg gccgagcgca gaagtgatcc tgcaacttta tccgcctcca tccagtctat
     5101 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     5161 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     5221 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     5281 ctccttcggt cctccgatcg ttgccagaag taagttggcc gcagtgttat cactcatggt
     5341 tatggcagca ctgcataatt ctcttactgt catgccatcc gtgagatgct tttctgtgac
     5401 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     5461 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     5521 tggaaaacgt tcttcggggc gtaaactctc aaggatctta ccgctgttga gatccagttc
     5581 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     5641 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     5701 atgttgaata ctcattttag cttccttagc tcctgaaaat ctcgataact caaaaaatac
     5761 gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt gccgatcaag
     5821 tcaaaagcct ccggtcggag gcttttgact ttctgctatg gaggtcaggt atgatttaaa
     5881 tggtcagtta ga
//
