LOCUS       sequence-298842-        7996 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 7996)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 7996)
  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..7996
                     /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             2857..2874
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2875..2892
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2937..2955)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             3306..4109
                     /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"
     terminator      6066..6095
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      6117..6704
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(6716..6743)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(6767..6958)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6959..7750,7751..7819))
                     /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     7582..7601
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7820..7922)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(7923..7954)
                     /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 cgtaacactg agagatcccc tcataatttc cccaaagcgt aaccatgtgt gaataaattt
      721 tgagctagta gggttgcagc cacgagtaag tcttcccttg ttattgtgta gccagaatgc
      781 cgcaaaactt ccatgcctaa gcgaactgtt gagagtacgt ttcgatttct gactgtgtta
      841 gcctggaagt gcttgtccca accttgtttc tgagcatgaa cgcccgcaag ccaacatgtt
      901 agttgaagca tcagggcgat tagcagcatg atatcaaaac gctctgagct gctcgttcgg
      961 ctatggcgta ggcctagtcc gtaggcagga cttttcaagt ctcggaaggt ttcttcaatc
     1021 tgcattcgct tcgaatagat attaacaagt tgtttgggtg ttcgaatttc aacaggtaag
     1081 ttagttgcta gaacccatgg ctcctttgcc gacgctgagt agattttagg tgacgggtgg
     1141 tgacaatgag tccgtgtcga gcgctgattt tttcggcctt tagagcgaga tttatacaat
     1201 agaatttggc atgagattgg attgctttta gtcagcctct tatagcctaa agtctttgag
     1261 tgactagatg acatatcatg taagttgctg ataggtttcc agttttccgc tcctaggtct
     1321 gcatattgta cttttcctct tactcgactt aaccagtacc aacccagctt ctcaacggat
     1381 ttataccatg gcactttaaa gccagcatca ctgacaatga gcggtgtggt gttactcggt
     1441 agaatgctcg caaggtcggc tagaaattgg tcatgagctt tctttgaaca ttgctctgaa
     1501 agcgggaacg ctttctcata aagagtaaca gaacgaccgt gtagtgcgac tgaagctcgc
     1561 aataccataa gtcgtttttg ctcacgaata tcagaccagt caacaagtac aatgggcatc
     1621 gtattgcccg aacagataaa gctagcatgc caacggtata cagcgagtcg ctctttgtgg
     1681 aggtgacgat tacctaacaa tcggtcgatt cgtttgatgt tatgttttgt tctcgctttg
     1741 gttggcaggt tacggccaag ttcggtaaga gtgagagttt tacagtcaag taatgcgtgg
     1801 caagccaacg ttaagctgtt gagtcgtttt aagtgtaatt cggggcagaa ttggtaaaga
     1861 gagtcgtgta aaatatcgag ttcgcacatc ttgttgtctg attattgatt tttcgcgaaa
     1921 ccatttgatc atatgacaag atgtgtatcc accttaactt aatgattttt accaaaatca
     1981 ttaggggatt catcagagcg taacatcaaa ccagtcccag aagttaagat tttgaacaat
     2041 ttgggtgttg acatcgctgc taataccgtc atttgggatt acaagagaga tgctccggcc
     2101 catatctcaa ctataggtgt ctgctcgatg actgacattg ctaaaaagcc aaccgagact
     2161 atctgtgctc cattaaccgt tttctttgac ggtagagtag atggtcaagt cgacctcttc
     2221 agaaacgcaa ggaacggtgt tttgattaca gaaggtagtg ttaaaggtct acaaccttct
     2281 gtaggtccaa agcaagcgtc cttgaacggc gtcacgttga ttggcgaagc cgttaagact
     2341 caattcaatt actataagaa ggtcgatggt gttgtgcaac aattaccaga aacttacttc
     2401 actcagtctc gcaatttgca agaatttaaa ccacgatctc aaatggaaat cgattttctt
     2461 gaactagcta tggacgagtt catcgaacgt tacaagttgg aaggttatgc ttttgaacac
     2521 attgtctacg gtgacttctc tcattcccag ctgggtggtt tacacttgct aattggactg
     2581 gctaaaagat tcaaggaatc accattcgag ttggaagatt ttatccctat ggactctacc
     2641 gttaaaaact acttcatcac tgatgctcaa accggttcga gtaagtgtgt ttgttccgtc
     2701 attgacttgt tactagatga tttcgtagaa ataattaagt ctcaagactt gtcagttgtt
     2761 tccaaggttg tcaaggtaac cattgactat acggaaatct catttatgtt gtggtgcaaa
     2821 gacggtcatg ttgaaacctt ctacccgaaa ttgcaactcg tgccgagagg ttctcaccat
     2881 catcaccatc attaaacagg ccccttttcc tttgtcgata tcatgtaatt agttatgtca
     2941 cgcttacatt cacgccctcc tcccacatcc gctctaaccg aaaaggaagg agttagacaa
     3001 cctgaagtct aggtccctat ttattttttt taatagttat gttagtatta agaacgttat
     3061 ttatatttca aatttttctt ttttttctgt acaaagcggc cgccgaacca ctagtgcact
     3121 gcagtacacg gtttccttga aatttttttg attcggtaat ctccgaacag aaggaagaac
     3181 gaaggaagga gcacagactt agattggtat atatacgcat atgtagtgtt gaagaaacat
     3241 gaaattgccc agtattctta acccaactgc acagaacaaa aacgtgcagg aaacgaagat
     3301 aaatcatgtc gaaagctaca tataaggaac gtgctgctac tcatcctagt cctgttgctg
     3361 ccaagctatt taatatcatg cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc
     3421 gtaccaccaa ggaattactg gagttagttg aagcattagg tcccaaaatt tgtttactaa
     3481 aaacacatgt ggatatcttg actgattttt ccatggaggg cacagttaag ccgctaaagg
     3541 cattatccgc caagtacaat tttttactct tcgaggacag aaaatttgct gacattggta
     3601 atacagtcaa attgcagtac tctgcgggtg tatacagaat agcagaatgg gcagacatta
     3661 cgaatgcaca cggtgtggtg ggcccaggta ttgttagcgg tttgaagcag gcggcagaag
     3721 aagtaacaaa ggaacctaga ggccttttga tgttagcaga attgtcatgc aagggctccc
     3781 tatctactgg agaatatact aagggtactg ttgacattgc gaagagcgac aaagattttg
     3841 ttatcggctt tattgctcaa agagacatgg gtggaagaga tgaaggttac gattggttga
     3901 ttatgacacc cggtgtgggt ttagatgaca agggagatgc attgggtcaa cagtatagaa
     3961 ccgtggatga tgtggtttct acaggatctg acattattat tgttggaaga ggactatttg
     4021 caaagggaag ggatgctaag gtagagggtg aacgttacag aaaagcaggc tgggaagcat
     4081 atttgagaag atgcggccag caaaactaaa aaactgtatt ataagtaaat gcatgtatac
     4141 taaactcaca aattagagct tcaatttaat tatatcagtt attacccgag taacgaagca
     4201 tctgtgcttc attttgtaga acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag
     4261 aatctgagct gcatttttac agaacagaaa tgcaacgcga aagcgctatt ttaccaacga
     4321 agaatctgtg cttcattttt gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa
     4381 caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgagagcg ctattttacc
     4441 aacaaagaat ctatacttct tttttgttct acaaaaatgc atcccgagag cgctattttt
     4501 ctaacaaagc atcttagatt actttttttc tcctttgtgc gctctataat gcagtctctt
     4561 gataactttt tgcactgtag gtccgttaag gttagaagaa ggctactttg gtgtctattt
     4621 tctcttccat aaaaaaagcc tgactccact tcccgcgttt actgattact agcgaagctg
     4681 cgggtgcatt ttttcaagat aaaggcatcc ccgattatat tctataccga tgtggattgc
     4741 gcatactttg tgaacagaaa gtgatagcgt tgatgattct tcattggtca gaaaattatg
     4801 aacggtttct tctattttgt ctctatatac tacgtatagg aaatgtttac attttcgtat
     4861 tgttttcgat tcactctatg aatggtcaac agtatagaac cgtggatgat gtggtttcta
     4921 caggatctga cattattatt gttggaagag gactatttgc aaagggaagg gatgctaagg
     4981 tagagggtga acgttacaga aaagcaggct gggaagcata tttgagaaga tgcggccagc
     5041 aaaactaaaa aactgtatta taagtaaatg catgtatact aaactcacaa attagagctt
     5101 caatttaatt atatcagtta ttacccgagt aacgaagcat ctgtgcttca ttttgtagaa
     5161 caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca
     5221 gaacagaaat gcaacgcgag agcgctattt taccaacaaa gaatctatac ttcttttttg
     5281 ttctacaaaa atgcatcccg agagcgctat ttttctaaca aagcatctta gattactttt
     5341 tttctccttt gtgcgctcta taatgcagtc tcttgataac tttttgcact gtaggtccgt
     5401 taaggttaga agaaggctac tttggtgtct attttctctt ccataaaaaa agcctgactc
     5461 cacttcccgc gtttactgat tactagcgaa gctgcgggtg cattttttca agataaaggc
     5521 atccccgatt atattctata ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata
     5581 gcgttgatga ttcttcattg gtcagaaaat tatgaacggt ttcttctatt ttgtctctat
     5641 atactacgta taggaaatgt ttacattttc gtattgtttt cgattcactc tatgaatagt
     5701 tcttactaca atttttttgt ctaaagagta atactagaga taaacataaa aaatgtagag
     5761 gtcgagttta gatgcaagtt caaggagcga aaggtggatg ggtaggttat atagggatat
     5821 agcacagaga tatatagcaa agagatactt ttgagcaatg tttgtggaag cggtattcgc
     5881 aatattttag tagctcgtta cagtccggtg cgtttttggt tttttgaaag tgcgtcatca
     5941 gagcgctttt ggttttcaaa agcgctctga agttcctata ctttctagct agagaatagg
     6001 aacttcccga gcggccgcgt gttacaagct tgactccagc gtaactggac tgcaatcaac
     6061 tcactggctc accttcacgg gtgggccttt cttcggtaga aaatcaaagg atcttcttga
     6121 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     6181 gtggtttgtt tgccggatca agagctacca actctttttc cgaggtaact ggcttcagca
     6241 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     6301 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     6361 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     6421 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     6481 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     6541 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     6601 caggggtaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     6661 atcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcag
     6721 aaaggcccac ccgaaggtga gccaggtgat tacatttggg ccctcatcag aggttttcac
     6781 cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg
     6841 attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta
     6901 atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactt
     6961 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     7021 ttgcctgact ccccgtcgtg tagataacta cgatgcggga gggcttacca tctggcccca
     7081 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     7141 agccagccgg gagggccgag cgcagaagtg atcctgcaac tttatccgcc tccatccagt
     7201 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     7261 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     7321 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     7381 ttagctcctt cggtcctccg atcgttgcca gaagtaagtt ggccgcagtg ttatcactca
     7441 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtgaga tgcttttctg
     7501 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     7561 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     7621 tcattggaaa acgttcttcg gggcgtaaac tctcaaggat cttaccgctg ttgagatcca
     7681 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     7741 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7801 ggaaatgttg aatactcatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa
     7861 atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat
     7921 caagtcaaaa gcctccggtc ggaggctttt gactttctgc tatggaggtc aggtatgatt
     7981 taaatggtca gttaga
//
