LOCUS       sequence-297808-        6351 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 6351)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6351)
  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-m4046653
FEATURES             Location/Qualifiers
     source          1..6351
                     /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"
     primer_bind     complement(2106..2124)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2475..3278
                     /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      3361..4343
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4421..4450
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4472..5059
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5071..5098)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5122..5313)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5314..6105,6106..6174))
                     /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     5937..5956
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6175..6277)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6278..6309)
                     /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 ataaacaggc cccttttcct ttgtcgatat catgtaatta
     2101 gttatgtcac gcttacattc acgccctcct cccacatccg ctctaaccga aaaggaagga
     2161 gttagacaac ctgaagtcta ggtccctatt tatttttttt aatagttatg ttagtattaa
     2221 gaacgttatt tatatttcaa atttttcttt tttttctgta caaagcggcc gccgaaccac
     2281 tagtgcactg cagtacacgg tttccttgaa atttttttga ttcggtaatc tccgaacaga
     2341 aggaagaacg aaggaaggag cacagactta gattggtata tatacgcata tgtagtgttg
     2401 aagaaacatg aaattgccca gtattcttaa cccaactgca cagaacaaaa acgtgcagga
     2461 aacgaagata aatcatgtcg aaagctacat ataaggaacg tgctgctact catcctagtc
     2521 ctgttgctgc caagctattt aatatcatgc acgaaaagca aacaaacttg tgtgcttcat
     2581 tggatgttcg taccaccaag gaattactgg agttagttga agcattaggt cccaaaattt
     2641 gtttactaaa aacacatgtg gatatcttga ctgatttttc catggagggc acagttaagc
     2701 cgctaaaggc attatccgcc aagtacaatt ttttactctt cgaggacaga aaatttgctg
     2761 acattggtaa tacagtcaaa ttgcagtact ctgcgggtgt atacagaata gcagaatggg
     2821 cagacattac gaatgcacac ggtgtggtgg gcccaggtat tgttagcggt ttgaagcagg
     2881 cggcagaaga agtaacaaag gaacctagag gccttttgat gttagcagaa ttgtcatgca
     2941 agggctccct atctactgga gaatatacta agggtactgt tgacattgcg aagagcgaca
     3001 aagattttgt tatcggcttt attgctcaaa gagacatggg tggaagagat gaaggttacg
     3061 attggttgat tatgacaccc ggtgtgggtt tagatgacaa gggagatgca ttgggtcaac
     3121 agtatagaac cgtggatgat gtggtttcta caggatctga cattattatt gttggaagag
     3181 gactatttgc aaagggaagg gatgctaagg tagagggtga acgttacaga aaagcaggct
     3241 gggaagcata tttgagaaga tgcggccagc aaaactaaaa aactgtatta taagtaaatg
     3301 catgtatact aaactcacaa attagagctt caatttaatt atatcagtta ttacccgagt
     3361 aacgaagcat ctgtgcttca ttttgtagaa caaaaatgca acgcgagagc gctaattttt
     3421 caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgaa agcgctattt
     3481 taccaacgaa gaatctgtgc ttcatttttg taaaacaaaa atgcaacgcg agagcgctaa
     3541 tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgagagcgc
     3601 tattttacca acaaagaatc tatacttctt ttttgttcta caaaaatgca tcccgagagc
     3661 gctatttttc taacaaagca tcttagatta ctttttttct cctttgtgcg ctctataatg
     3721 cagtctcttg ataacttttt gcactgtagg tccgttaagg ttagaagaag gctactttgg
     3781 tgtctatttt ctcttccata aaaaaagcct gactccactt cccgcgttta ctgattacta
     3841 gcgaagctgc gggtgcattt tttcaagata aaggcatccc cgattatatt ctataccgat
     3901 gtggattgcg catactttgt gaacagaaag tgatagcgtt gatgattctt cattggtcag
     3961 aaaattatga acggtttctt ctattttgtc tctatatact acgtatagga aatgtttaca
     4021 ttttcgtatt gttttcgatt cactctatga atagttctta ctacaatttt tttgtctaaa
     4081 gagtaatact agagataaac ataaaaaatg tagaggtcga gtttagatgc aagttcaagg
     4141 agcgaaaggt ggatgggtag gttatatagg gatatagcac agagatatat agcaaagaga
     4201 tacttttgag caatgtttgt ggaagcggta ttcgcaatat tttagtagct cgttacagtc
     4261 cggtgcgttt ttggtttttt gaaagtgcgt catcagagcg cttttggttt tcaaaagcgc
     4321 tctgaagttc ctatactttc tagctagaga ataggaactt cccgagcggc cgcgtgttac
     4381 aagcttgact ccagcgtaac tggactgcaa tcaactcact ggctcacctt cacgggtggg
     4441 cctttcttcg gtagaaaatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     4501 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     4561 taccaactct ttttccgagg taactggctt cagcagagcg cagataccaa atactgttct
     4621 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     4681 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     4741 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     4801 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     4861 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     4921 cagggtcgga acaggagagc gcacgaggga gcttccaggg gtaaacgcct ggtatcttta
     4981 tagtcctgtc gggtttcgcc acctctgact tgagcatcga tttttgtgat gctcgtcagg
     5041 ggggcggagc ctatggaaaa acgccagcaa cgcagaaagg cccacccgaa ggtgagccag
     5101 gtgattacat ttgggccctc atcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca
     5161 gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc
     5221 cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc
     5281 catgttaagg gcggtttttt cctgtttggt cacttaccaa tgcttaatca gtgaggcacc
     5341 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     5401 aactacgatg cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     5461 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccgggaggg ccgagcgcag
     5521 aagtgatcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     5581 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5641 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5701 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5761 tgccagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5821 tcttactgtc atgccatccg tgagatgctt ttctgtgact ggtgagtact caaccaagtc
     5881 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     5941 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     6001 taaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     6061 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     6121 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcattttagc
     6181 ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg atcttatttc
     6241 attatggtga aagttggaac ctcttacgtg ccgatcaagt caaaagcctc cggtcggagg
     6301 cttttgactt tctgctatgg aggtcaggta tgatttaaat ggtcagttag a
//
