LOCUS       sequence-297877-        6381 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 3'-to-5' exonuclease.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6381)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6381)
  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-m4046242
FEATURES             Location/Qualifiers
     source          1..6381
                     /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             2056..2073
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2074..2091
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2136..2154)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2505..3308
                     /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      3391..4373
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4451..4480
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4502..5089
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5101..5128)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5152..5343)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5344..6135,6136..6204))
                     /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     5967..5986
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6205..6307)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6308..6339)
                     /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 gaaaatgtaa caggactctt taaagattgt agtaaggtaa tcactgggtt acatcctaca
      541 caggcaccta cacacctcag tgttgacact aaattcaaaa ctgaaggttt atgtgttgac
      601 atacctggca tacctaagga catgacctat agaagactca tctctatgat gggttttaaa
      661 atgaattatc aagttaatgg ttaccctaac atgtttatca cccgcgaaga agctataaga
      721 catgtacgtg catggattgg cttcgatgtc gaggggtgtc atgctactag agaagctgtt
      781 ggtaccaatt tacctttaca gctaggtttt tctacaggtg ttaacctagt tgctgtacct
      841 acaggttatg ttgatacacc taataataca gatttttcca gagttagtgc taaaccaccg
      901 cctggagatc aatttaaaca cctcatacca cttatgtaca aaggacttcc ttggaatgta
      961 gtgcgtataa agattgtaca aatgttaagt gacacactta aaaatctctc tgacagagtc
     1021 gtatttgtct tatgggcaca tggctttgag ttgacatcta tgaagtattt tgtgaaaata
     1081 ggacctgagc gcacctgttg tctatgtgat agacgtgcca catgcttttc cactgcttca
     1141 gacacttatg cctgttggca tcattctatt ggatttgatt acgtctataa tccgtttatg
     1201 attgatgttc aacaatgggg ttttacaggt aacctacaaa gcaaccatga tctgtattgt
     1261 caagtccatg gtaatgcaca tgtagctagt tgtgatgcaa tcatgactag gtgtctagct
     1321 gtccacgagt gctttgttaa gcgtgttgac tggactattg aatatcctat aattggtgat
     1381 gaactgaaga ttaatgcggc ttgtagaaag gttcaacaca tggttgttaa agctgcatta
     1441 ttagcagaca aattcccagt tcttcacgac attggtaacc ctaaagctat taagtgtgta
     1501 cctcaagctg atgtagaatg gaagttctat gatgcacagc cttgtagtga caaagcttat
     1561 aaaatagaag aattattcta ttcttatgcc acacattctg acaaattcac agatggtgta
     1621 tgcctatttt ggaattgcaa tgtcgataga tatcctgcta attccattgt ttgtagattt
     1681 gacactagag tgctatctaa ccttaacttg cctggttgtg atggtggcag tttgtatgta
     1741 aataaacatg cattccacac accagctttt gataaaagtg cttttgttaa tttaaaacaa
     1801 ttaccatttt tctattactc tgacagtcca tgtgagtctc atggaaaaca agtagtgtca
     1861 gatatagatt atgtaccact aaagtctgct acgtgtataa cacgttgcaa tttaggtggt
     1921 gctgtctgta gacatcatgc taatgagtac agattgtatc tcgatgctta taacatgatg
     1981 atctcagctg gctttagctt gtgggtttac aaacaatttg atacttataa cctctggaac
     2041 acttttacaa gacttctcgt gccgagaggt tctcaccatc atcaccatca ttaaacaggc
     2101 cccttttcct ttgtcgatat catgtaatta gttatgtcac gcttacattc acgccctcct
     2161 cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta ggtccctatt
     2221 tatttttttt aatagttatg ttagtattaa gaacgttatt tatatttcaa atttttcttt
     2281 tttttctgta caaagcggcc gccgaaccac tagtgcactg cagtacacgg tttccttgaa
     2341 atttttttga ttcggtaatc tccgaacaga aggaagaacg aaggaaggag cacagactta
     2401 gattggtata tatacgcata tgtagtgttg aagaaacatg aaattgccca gtattcttaa
     2461 cccaactgca cagaacaaaa acgtgcagga aacgaagata aatcatgtcg aaagctacat
     2521 ataaggaacg tgctgctact catcctagtc ctgttgctgc caagctattt aatatcatgc
     2581 acgaaaagca aacaaacttg tgtgcttcat tggatgttcg taccaccaag gaattactgg
     2641 agttagttga agcattaggt cccaaaattt gtttactaaa aacacatgtg gatatcttga
     2701 ctgatttttc catggagggc acagttaagc cgctaaaggc attatccgcc aagtacaatt
     2761 ttttactctt cgaggacaga aaatttgctg acattggtaa tacagtcaaa ttgcagtact
     2821 ctgcgggtgt atacagaata gcagaatggg cagacattac gaatgcacac ggtgtggtgg
     2881 gcccaggtat tgttagcggt ttgaagcagg cggcagaaga agtaacaaag gaacctagag
     2941 gccttttgat gttagcagaa ttgtcatgca agggctccct atctactgga gaatatacta
     3001 agggtactgt tgacattgcg aagagcgaca aagattttgt tatcggcttt attgctcaaa
     3061 gagacatggg tggaagagat gaaggttacg attggttgat tatgacaccc ggtgtgggtt
     3121 tagatgacaa gggagatgca ttgggtcaac agtatagaac cgtggatgat gtggtttcta
     3181 caggatctga cattattatt gttggaagag gactatttgc aaagggaagg gatgctaagg
     3241 tagagggtga acgttacaga aaagcaggct gggaagcata tttgagaaga tgcggccagc
     3301 aaaactaaaa aactgtatta taagtaaatg catgtatact aaactcacaa attagagctt
     3361 caatttaatt atatcagtta ttacccgagt aacgaagcat ctgtgcttca ttttgtagaa
     3421 caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca
     3481 gaacagaaat gcaacgcgaa agcgctattt taccaacgaa gaatctgtgc ttcatttttg
     3541 taaaacaaaa atgcaacgcg agagcgctaa tttttcaaac aaagaatctg agctgcattt
     3601 ttacagaaca gaaatgcaac gcgagagcgc tattttacca acaaagaatc tatacttctt
     3661 ttttgttcta caaaaatgca tcccgagagc gctatttttc taacaaagca tcttagatta
     3721 ctttttttct cctttgtgcg ctctataatg cagtctcttg ataacttttt gcactgtagg
     3781 tccgttaagg ttagaagaag gctactttgg tgtctatttt ctcttccata aaaaaagcct
     3841 gactccactt cccgcgttta ctgattacta gcgaagctgc gggtgcattt tttcaagata
     3901 aaggcatccc cgattatatt ctataccgat gtggattgcg catactttgt gaacagaaag
     3961 tgatagcgtt gatgattctt cattggtcag aaaattatga acggtttctt ctattttgtc
     4021 tctatatact acgtatagga aatgtttaca ttttcgtatt gttttcgatt cactctatga
     4081 atagttctta ctacaatttt tttgtctaaa gagtaatact agagataaac ataaaaaatg
     4141 tagaggtcga gtttagatgc aagttcaagg agcgaaaggt ggatgggtag gttatatagg
     4201 gatatagcac agagatatat agcaaagaga tacttttgag caatgtttgt ggaagcggta
     4261 ttcgcaatat tttagtagct cgttacagtc cggtgcgttt ttggtttttt gaaagtgcgt
     4321 catcagagcg cttttggttt tcaaaagcgc tctgaagttc ctatactttc tagctagaga
     4381 ataggaactt cccgagcggc cgcgtgttac aagcttgact ccagcgtaac tggactgcaa
     4441 tcaactcact ggctcacctt cacgggtggg cctttcttcg gtagaaaatc aaaggatctt
     4501 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     4561 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgagg taactggctt
     4621 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     4681 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     4741 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     4801 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     4861 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     4921 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     4981 gcttccaggg gtaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     5041 tgagcatcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     5101 cgcagaaagg cccacccgaa ggtgagccag gtgattacat ttgggccctc atcagaggtt
     5161 ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg
     5221 aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag
     5281 cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt
     5341 cacttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     5401 catagttgcc tgactccccg tcgtgtagat aactacgatg cgggagggct taccatctgg
     5461 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     5521 aaaccagcca gccgggaggg ccgagcgcag aagtgatcct gcaactttat ccgcctccat
     5581 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     5641 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     5701 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     5761 agcggttagc tccttcggtc ctccgatcgt tgccagaagt aagttggccg cagtgttatc
     5821 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg tgagatgctt
     5881 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     5941 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     6001 gctcatcatt ggaaaacgtt cttcggggcg taaactctca aggatcttac cgctgttgag
     6061 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     6121 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     6181 gacacggaaa tgttgaatac tcattttagc ttccttagct cctgaaaatc tcgataactc
     6241 aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac ctcttacgtg
     6301 ccgatcaagt caaaagcctc cggtcggagg cttttgactt tctgctatgg aggtcaggta
     6361 tgatttaaat ggtcagttag a
//
