LOCUS       sequence-297715-        7461 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 7461)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 7461)
  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-m4046344
FEATURES             Location/Qualifiers
     source          1..7461
                     /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"
     primer_bind     complement(3216..3234)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             3585..4388
                     /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      4471..5453
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      5531..5560
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      5582..6169
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(6181..6208)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(6232..6423)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6424..7215,7216..7284))
                     /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     7047..7066
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7285..7387)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(7388..7419)
                     /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 tctaaaatcg aggaaggtaa gctagtcatt
     2101 tggataaatg gagataaggg ttacaacggt ttggcagaag taggcaagaa atttgaaaag
     2161 gacacaggta tcaaggtgac ggttgaacat ccagacaagt tagaagaaaa attccctcaa
     2221 gttgctgcta ctggtgatgg tcccgacatt attttctggg cccacgatcg ttttggtgga
     2281 tatgctcagt caggtttgct ggctgagatc accccagaca aggccttcca agataaattg
     2341 tacccattca cttgggatgc tgtcagatac aacggtaagc ttatagcata tccaattgct
     2401 gttgaagcct tatcgttgat ctacaataag gacttgttac ctaacccacc aaagacatgg
     2461 gaagaaattc cagctttgga taaagagcta aaggccaagg gaaaaagcgc tttgatgttt
     2521 aacttacaag aaccttattt cacctggcca ttaatcgctg ctgacggtgg ttacgcattt
     2581 aaatacgaaa acggtaagta tgacattaag gatgtcggcg ttgataatgc tggtgcaaag
     2641 gctggtttga ccttcttggt tgatttaata aagaataaac acatgaacgc tgacactgac
     2701 tactctatcg ccgaagccgc tttcaacaag ggtgaaactg ctatgaccat aaatggtcca
     2761 tgggcctgga gtaacattga tacttctaaa gtaaactatg gtgtcactgt tttgcctaca
     2821 ttcaaaggcc aaccatccaa gccctttgtc ggtgttctat ccgctggcat caatgctgct
     2881 tctccaaaca aggagctagc taaagaattt ttggaaaatt acttgttaac tgacgaaggt
     2941 ttggaggccg ttaacaagga caagccattg ggtgctgtcg ctttgaaatc ttacgaagaa
     3001 gaattggcta aggatcccag aattgccgct accatggaga acgcccaaaa gggtgaaatt
     3061 atgccaaaca tcccacaaat gtcagccttc tggtacgcag ttaggactgc cgtgattaat
     3121 gctgcttccg gtagacaaac cgtcgacgaa gcattaaaag atgctcaaac gtaaacaggc
     3181 cccttttcct ttgtcgatat catgtaatta gttatgtcac gcttacattc acgccctcct
     3241 cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta ggtccctatt
     3301 tatttttttt aatagttatg ttagtattaa gaacgttatt tatatttcaa atttttcttt
     3361 tttttctgta caaagcggcc gccgaaccac tagtgcactg cagtacacgg tttccttgaa
     3421 atttttttga ttcggtaatc tccgaacaga aggaagaacg aaggaaggag cacagactta
     3481 gattggtata tatacgcata tgtagtgttg aagaaacatg aaattgccca gtattcttaa
     3541 cccaactgca cagaacaaaa acgtgcagga aacgaagata aatcatgtcg aaagctacat
     3601 ataaggaacg tgctgctact catcctagtc ctgttgctgc caagctattt aatatcatgc
     3661 acgaaaagca aacaaacttg tgtgcttcat tggatgttcg taccaccaag gaattactgg
     3721 agttagttga agcattaggt cccaaaattt gtttactaaa aacacatgtg gatatcttga
     3781 ctgatttttc catggagggc acagttaagc cgctaaaggc attatccgcc aagtacaatt
     3841 ttttactctt cgaggacaga aaatttgctg acattggtaa tacagtcaaa ttgcagtact
     3901 ctgcgggtgt atacagaata gcagaatggg cagacattac gaatgcacac ggtgtggtgg
     3961 gcccaggtat tgttagcggt ttgaagcagg cggcagaaga agtaacaaag gaacctagag
     4021 gccttttgat gttagcagaa ttgtcatgca agggctccct atctactgga gaatatacta
     4081 agggtactgt tgacattgcg aagagcgaca aagattttgt tatcggcttt attgctcaaa
     4141 gagacatggg tggaagagat gaaggttacg attggttgat tatgacaccc ggtgtgggtt
     4201 tagatgacaa gggagatgca ttgggtcaac agtatagaac cgtggatgat gtggtttcta
     4261 caggatctga cattattatt gttggaagag gactatttgc aaagggaagg gatgctaagg
     4321 tagagggtga acgttacaga aaagcaggct gggaagcata tttgagaaga tgcggccagc
     4381 aaaactaaaa aactgtatta taagtaaatg catgtatact aaactcacaa attagagctt
     4441 caatttaatt atatcagtta ttacccgagt aacgaagcat ctgtgcttca ttttgtagaa
     4501 caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca
     4561 gaacagaaat gcaacgcgaa agcgctattt taccaacgaa gaatctgtgc ttcatttttg
     4621 taaaacaaaa atgcaacgcg agagcgctaa tttttcaaac aaagaatctg agctgcattt
     4681 ttacagaaca gaaatgcaac gcgagagcgc tattttacca acaaagaatc tatacttctt
     4741 ttttgttcta caaaaatgca tcccgagagc gctatttttc taacaaagca tcttagatta
     4801 ctttttttct cctttgtgcg ctctataatg cagtctcttg ataacttttt gcactgtagg
     4861 tccgttaagg ttagaagaag gctactttgg tgtctatttt ctcttccata aaaaaagcct
     4921 gactccactt cccgcgttta ctgattacta gcgaagctgc gggtgcattt tttcaagata
     4981 aaggcatccc cgattatatt ctataccgat gtggattgcg catactttgt gaacagaaag
     5041 tgatagcgtt gatgattctt cattggtcag aaaattatga acggtttctt ctattttgtc
     5101 tctatatact acgtatagga aatgtttaca ttttcgtatt gttttcgatt cactctatga
     5161 atagttctta ctacaatttt tttgtctaaa gagtaatact agagataaac ataaaaaatg
     5221 tagaggtcga gtttagatgc aagttcaagg agcgaaaggt ggatgggtag gttatatagg
     5281 gatatagcac agagatatat agcaaagaga tacttttgag caatgtttgt ggaagcggta
     5341 ttcgcaatat tttagtagct cgttacagtc cggtgcgttt ttggtttttt gaaagtgcgt
     5401 catcagagcg cttttggttt tcaaaagcgc tctgaagttc ctatactttc tagctagaga
     5461 ataggaactt cccgagcggc cgcgtgttac aagcttgact ccagcgtaac tggactgcaa
     5521 tcaactcact ggctcacctt cacgggtggg cctttcttcg gtagaaaatc aaaggatctt
     5581 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     5641 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgagg taactggctt
     5701 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     5761 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5821 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5881 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5941 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     6001 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     6061 gcttccaggg gtaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     6121 tgagcatcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     6181 cgcagaaagg cccacccgaa ggtgagccag gtgattacat ttgggccctc atcagaggtt
     6241 ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg
     6301 aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag
     6361 cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt
     6421 cacttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     6481 catagttgcc tgactccccg tcgtgtagat aactacgatg cgggagggct taccatctgg
     6541 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     6601 aaaccagcca gccgggaggg ccgagcgcag aagtgatcct gcaactttat ccgcctccat
     6661 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     6721 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     6781 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     6841 agcggttagc tccttcggtc ctccgatcgt tgccagaagt aagttggccg cagtgttatc
     6901 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg tgagatgctt
     6961 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     7021 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     7081 gctcatcatt ggaaaacgtt cttcggggcg taaactctca aggatcttac cgctgttgag
     7141 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     7201 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     7261 gacacggaaa tgttgaatac tcattttagc ttccttagct cctgaaaatc tcgataactc
     7321 aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac ctcttacgtg
     7381 ccgatcaagt caaaagcctc cggtcggagg cttttgactt tctgctatgg aggtcaggta
     7441 tgatttaaat ggtcagttag a
//
