LOCUS       sequence-298140-        6361 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 3'-to-5' exonuclease.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6361)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6361)
  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-m4046374
FEATURES             Location/Qualifiers
     source          1..6361
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     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             2062..2079
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2080..2097
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2142..2160)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2485..3288
                     /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      3371..4353
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4431..4460
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4482..5069
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5081..5108)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5132..5323)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5324..6115,6116..6184))
                     /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     5947..5966
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6185..6287)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6288..6319)
                     /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 gccgagaatg tcacaggcct attcaaggat tgctcgaaag tgatcaccgg tttacaccct
      541 acgcaggctc caactcattt gtccgttgac accaagttta agactgaagg attgtgtgtt
      601 gacataccag gtattcccaa agatatgact taccgtaggc tgatttctat gatgggtttc
      661 aagatgaact atcaagttaa cggttaccca aatatgttta tcaccagaga agaagctatt
      721 agacacgtac gtgcatggat cggtttcgat gtcgaaggtt gtcatgctac tagagaagcg
      781 gtcggtacaa acttgccatt acaacttggc ttttcaaccg gtgttaattt ggttgctgtt
      841 cctacagggt atgtcgacac tccaaacaac acggatttct ctcgagtgtc cgccaagcca
      901 ccacctggtg atcaattcaa acacttgatc ccgctcatgt acaagggttt gccttggaac
      961 gttgtcagaa ttaaaatagt acaaatgtta tctgatactt tgaagaatct gagtgacaga
     1021 gttgtcttcg tcctttgggc tcacggtttt gagctaacca gcatgaagta ctttgtgaag
     1081 attggaccag aacggacttg ttgcttgtgt gacagaagag ccacttgttt ctccaccgct
     1141 agcgatacat atgcatgttg gcatcattct attggtttcg actacgttta caaccccttt
     1201 atgatcgacg ttcaacagtg gggtttcacc ggtaatttgc aatcgaacca cgacttgtat
     1261 tgtcaagttc acggcaatgc tcacgtggct tcatgcgatg ctattatgac aaggtgttta
     1321 gccgtccatg aatgtttcgt taaaagagta gactggacta tcgaataccc aattatcggt
     1381 gatgaactca agattaacgc tgcctgtaga aaggtccaac atatggttgt aaaggcagct
     1441 ctattagctg acaaatttcc agttttgcac gatataggga atccaaaggc cataaagtgt
     1501 gttccacaag ctgacgtcga atggaaattc tacgatgcgc aaccctgctc tgataaagct
     1561 tataagatcg aggaattgtt ttactcttac gctacccact ccgataagtt cactgacggt
     1621 gtgtgtcttt tctggaactg caatgttgat agatatccag caaactccat cgtttgtaga
     1681 ttcgacacgc gagtcttgag caacttgaac ttaccaggtt gtgatggtgg ttccttgtac
     1741 gtcaataaac atgcatttca cactcccgct ttcgacaagt ctgcttttgt taatttaaag
     1801 caattgcctt ttttctatta ctctgatagt ccatgcgaat ctcatggtaa gcaagtcgtt
     1861 tccgacattg attacgtacc actgaagtca gccacctgta taacaagatg taaccttgga
     1921 ggtgctgtct gccgccacca tgctaacgag tacagactat acttggatgc ctataacatg
     1981 atgatttcag ccggtttctc tttatgggtt tacaagcagt ttgacactta caacttgtgg
     2041 aacaccttca cgcgtttgca actcgtgccg agaggttctc accatcatca ccatcattaa
     2101 acaggcccct tttcctttgt cgatatcatg taattagtta tgtcacgctt acattcacgc
     2161 cctcctccca catccgctct aaccgaaaag gaaggagtta gacaacctga agtctaggtc
     2221 cctatttatt ttttttaata gttatttcaa atttttcttt tttttctgta caaagcggcc
     2281 gccgaaccac tagtgcactg cagtacacgg tttccttgaa atttttttga ttcggtaatc
     2341 tccgaacaga aggaagaacg aaggaaggag cacagactta gattggtata tatacgcata
     2401 tgtagtgttg aagaaacatg aaattgccca gtattcttaa cccaactgca cagaacaaaa
     2461 acgtgcagga aacgaagata aatcatgtcg aaagctacat ataaggaacg tgctgctact
     2521 catcctagtc ctgttgctgc caagctattt aatatcatgc acgaaaagca aacaaacttg
     2581 tgtgcttcat tggatgttcg taccaccaag gaattactgg agttagttga agcattaggt
     2641 cccaaaattt gtttactaaa aacacatgtg gatatcttga ctgatttttc catggagggc
     2701 acagttaagc cgctaaaggc attatccgcc aagtacaatt ttttactctt cgaggacaga
     2761 aaatttgctg acattggtaa tacagtcaaa ttgcagtact ctgcgggtgt atacagaata
     2821 gcagaatggg cagacattac gaatgcacac ggtgtggtgg gcccaggtat tgttagcggt
     2881 ttgaagcagg cggcagaaga agtaacaaag gaacctagag gccttttgat gttagcagaa
     2941 ttgtcatgca agggctccct atctactgga gaatatacta agggtactgt tgacattgcg
     3001 aagagcgaca aagattttgt tatcggcttt attgctcaaa gagacatggg tggaagagat
     3061 gaaggttacg attggttgat tatgacaccc ggtgtgggtt tagatgacaa gggagatgca
     3121 ttgggtcaac agtatagaac cgtggatgat gtggtttcta caggatctga cattattatt
     3181 gttggaagag gactatttgc aaagggaagg gatgctaagg tagagggtga acgttacaga
     3241 aaagcaggct gggaagcata tttgagaaga tgcggccagc aaaactaaaa aactgtatta
     3301 taagtaaatg catgtatact aaactcacaa attagagctt caatttaatt atatcagtta
     3361 ttacccgagt aacgaagcat ctgtgcttca ttttgtagaa caaaaatgca acgcgagagc
     3421 gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgaa
     3481 agcgctattt taccaacgaa gaatctgtgc ttcatttttg taaaacaaaa atgcaacgcg
     3541 agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac
     3601 gcgagagcgc tattttacca acaaagaatc tatacttctt ttttgttcta caaaaatgca
     3661 tcccgagagc gctatttttc taacaaagca tcttagatta ctttttttct cctttgtgcg
     3721 ctctataatg cagtctcttg ataacttttt gcactgtagg tccgttaagg ttagaagaag
     3781 gctactttgg tgtctatttt ctcttccata aaaaaagcct gactccactt cccgcgttta
     3841 ctgattacta gcgaagctgc gggtgcattt tttcaagata aaggcatccc cgattatatt
     3901 ctataccgat gtggattgcg catactttgt gaacagaaag tgatagcgtt gatgattctt
     3961 cattggtcag aaaattatga acggtttctt ctattttgtc tctatatact acgtatagga
     4021 aatgtttaca ttttcgtatt gttttcgatt cactctatga atagttctta ctacaatttt
     4081 tttgtctaaa gagtaatact agagataaac ataaaaaatg tagaggtcga gtttagatgc
     4141 aagttcaagg agcgaaaggt ggatgggtag gttatatagg gatatagcac agagatatat
     4201 agcaaagaga tacttttgag caatgtttgt ggaagcggta ttcgcaatat tttagtagct
     4261 cgttacagtc cggtgcgttt ttggtttttt gaaagtgcgt catcagagcg cttttggttt
     4321 tcaaaagcgc tctgaagttc ctatactttc tagctagaga ataggaactt cccgagcggc
     4381 cgcgtgttac aagcttgact ccagcgtaac tggactgcaa tcaactcact ggctcacctt
     4441 cacgggtggg cctttcttcg gtagaaaatc aaaggatctt cttgagatcc tttttttctg
     4501 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4561 gatcaagagc taccaactct ttttccgagg taactggctt cagcagagcg cagataccaa
     4621 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     4681 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     4741 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     4801 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     4861 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     4921 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg gtaaacgcct
     4981 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcatcga tttttgtgat
     5041 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcagaaagg cccacccgaa
     5101 ggtgagccag gtgattacat ttgggccctc atcagaggtt ttcaccgtca tcaccgaaac
     5161 gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg
     5221 cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga
     5281 taaagcgggc catgttaagg gcggtttttt cctgtttggt cacttaccaa tgcttaatca
     5341 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     5401 tcgtgtagat aactacgatg cgggagggct taccatctgg ccccagtgct gcaatgatac
     5461 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccgggaggg
     5521 ccgagcgcag aagtgatcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     5581 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     5641 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5701 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5761 ctccgatcgt tgccagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5821 tgcataattc tcttactgtc atgccatccg tgagatgctt ttctgtgact ggtgagtact
     5881 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5941 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     6001 cttcggggcg taaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     6061 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     6121 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     6181 tcattttagc ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg
     6241 atcnnatttc attatggtga aagttggaac ctcttacgtg ccgatcaagt caaaagcctc
     6301 cggtcggagg cttttgactt tctgctatgg aggtcaggta tgatttaaat ggtcagttag
     6361 a
//
