LOCUS       sequence-287745-        5844 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 5844)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5844)
  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-m4046438
FEATURES             Location/Qualifiers
     source          1..5844
                     /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..1554
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1599..1617)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1968..2771
                     /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      2854..3836
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3914..3943
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3965..4552
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4564..4591)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4615..4806)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4807..5598,5599..5667))
                     /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     5430..5449
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5668..5770)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5771..5802)
                     /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 ggttctcacc atcatcacca tcattaaaca
     1561 ggcccctttt cctttgtcga tatcatgtaa ttagttatgt cacgcttaca ttcacgccct
     1621 cctcccacat ccgctctaac cgaaaaggaa ggagttagac aacctgaagt ctaggtccct
     1681 atttattttt tttaatagtt atgttagtat taagaacgtt atttatattt caaatttttc
     1741 ttttttttct gtacaaagcg gccgccgaac cactagtgca ctgcagtaca cggtttcctt
     1801 gaaatttttt tgattcggta atctccgaac agaaggaaga acgaaggaag gagcacagac
     1861 ttagattggt atatatacgc atatgtagtg ttgaagaaac atgaaattgc ccagtattct
     1921 taacccaact gcacagaaca aaaacgtgca ggaaacgaag ataaatcatg tcgaaagcta
     1981 catataagga acgtgctgct actcatccta gtcctgttgc tgccaagcta tttaatatca
     2041 tgcacgaaaa gcaaacaaac ttgtgtgctt cattggatgt tcgtaccacc aaggaattac
     2101 tggagttagt tgaagcatta ggtcccaaaa tttgtttact aaaaacacat gtggatatct
     2161 tgactgattt ttccatggag ggcacagtta agccgctaaa ggcattatcc gccaagtaca
     2221 attttttact cttcgaggac agaaaatttg ctgacattgg taatacagtc aaattgcagt
     2281 actctgcggg tgtatacaga atagcagaat gggcagacat tacgaatgca cacggtgtgg
     2341 tgggcccagg tattgttagc ggtttgaagc aggcggcaga agaagtaaca aaggaaccta
     2401 gaggcctttt gatgttagca gaattgtcat gcaagggctc cctatctact ggagaatata
     2461 ctaagggtac tgttgacatt gcgaagagcg acaaagattt tgttatcggc tttattgctc
     2521 aaagagacat gggtggaaga gatgaaggtt acgattggtt gattatgaca cccggtgtgg
     2581 gtttagatga caagggagat gcattgggtc aacagtatag aaccgtggat gatgtggttt
     2641 ctacaggatc tgacattatt attgttggaa gaggactatt tgcaaaggga agggatgcta
     2701 aggtagaggg tgaacgttac agaaaagcag gctgggaagc atatttgaga agatgcggcc
     2761 agcaaaacta aaaaactgta ttataagtaa atgcatgtat actaaactca caaattagag
     2821 cttcaattta attatatcag ttattacccg agtaacgaag catctgtgct tcattttgta
     2881 gaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt
     2941 acagaacaga aatgcaacgc gaaagcgcta ttttaccaac gaagaatctg tgcttcattt
     3001 ttgtaaaaca aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca
     3061 tttttacaga acagaaatgc aacgcgagag cgctatttta ccaacaaaga atctatactt
     3121 cttttttgtt ctacaaaaat gcatcccgag agcgctattt ttctaacaaa gcatcttaga
     3181 ttactttttt tctcctttgt gcgctctata atgcagtctc ttgataactt tttgcactgt
     3241 aggtccgtta aggttagaag aaggctactt tggtgtctat tttctcttcc ataaaaaaag
     3301 cctgactcca cttcccgcgt ttactgatta ctagcgaagc tgcgggtgca ttttttcaag
     3361 ataaaggcat ccccgattat attctatacc gatgtggatt gcgcatactt tgtgaacaga
     3421 aagtgatagc gttgatgatt cttcattggt cagaaaatta tgaacggttt cttctatttt
     3481 gtctctatat actacgtata ggaaatgttt acattttcgt attgttttcg attcactcta
     3541 tgaatagttc ttactacaat ttttttgtct aaagagtaat actagagata aacataaaaa
     3601 atgtagaggt cgagtttaga tgcaagttca aggagcgaaa ggtggatggg taggttatat
     3661 agggatatag cacagagata tatagcaaag agatactttt gagcaatgtt tgtggaagcg
     3721 gtattcgcaa tattttagta gctcgttaca gtccggtgcg tttttggttt tttgaaagtg
     3781 cgtcatcaga gcgcttttgg ttttcaaaag cgctctgaag ttcctatact ttctagctag
     3841 agaataggaa cttcccgagc ggccgcgtgt tacaagcttg actccagcgt aactggactg
     3901 caatcaactc actggctcac cttcacgggt gggcctttct tcggtagaaa atcaaaggat
     3961 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     4021 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aggtaactgg
     4081 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     4141 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4201 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4261 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     4321 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     4381 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     4441 ggagcttcca ggggtaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     4501 acttgagcat cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     4561 caacgcagaa aggcccaccc gaaggtgagc caggtgatta catttgggcc ctcatcagag
     4621 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
     4681 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
     4741 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
     4801 ggtcacttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     4861 atccatagtt gcctgactcc ccgtcgtgta gataactacg atgcgggagg gcttaccatc
     4921 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     4981 aataaaccag ccagccggga gggccgagcg cagaagtgat cctgcaactt tatccgcctc
     5041 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5101 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5161 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     5221 aaaagcggtt agctccttcg gtcctccgat cgttgccaga agtaagttgg ccgcagtgtt
     5281 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtgagatg
     5341 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5401 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5461 agtgctcatc attggaaaac gttcttcggg gcgtaaactc tcaaggatct taccgctgtt
     5521 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5581 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     5641 ggcgacacgg aaatgttgaa tactcatttt agcttcctta gctcctgaaa atctcgataa
     5701 ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg aacctcttac
     5761 gtgccgatca agtcaaaagc ctccggtcgg aggcttttga ctttctgcta tggaggtcag
     5821 gtatgattta aatggtcagt taga
//
