LOCUS       sequence-297566-        7414 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 7414)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 7414)
  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-m4046289
FEATURES             Location/Qualifiers
     source          1..7414
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             2009..2026
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(3169..3187)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             3538..4341
                     /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      4424..5406
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      5484..5513
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      5535..6122
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(6134..6161)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(6185..6376)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6377..7168,7169..7237))
                     /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     7000..7019
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7238..7340)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(7341..7372)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgccc cacacaccat agcttcaaaa tgtttctact ccttttttac tcttccagat
       61 tttctcggac tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag catactaaat
      121 ttcccctctt tcttcctcta gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa
      181 aaaagagacc gcctcgtttc tttttcttcg tcgaaaaagg caataaaaat ttttatcacg
      241 tttctttttc ttgaaaattt ttttttttga tttttttctc tttcgatgac ctcccattga
      301 tatttaagtt aataaacggt cttcaatttc tcaagtttca gtttcatttt tcttgttcta
      361 ttacaacttt ttttacttct tgctcattag aaagaaagca tagcaatcta atctaagttt
      421 taattacaaa atggaaaatg taacaggact ctttaaagat tgtagtaagg taatcactgg
      481 gttacatcct acacaggcac ctacacacct cagtgttgac actaaattca aaactgaagg
      541 tttatgtgtt gacatacctg gcatacctaa ggacatgacc tatagaagac tcatctctat
      601 gatgggtttt aaaatgaatt atcaagttaa tggttaccct aacatgttta tcacccgcga
      661 agaagctata agacatgtac gtgcatggat tggcttcgat gtcgaggggt gtcatgctac
      721 tagagaagct gttggtacca atttaccttt acagctaggt ttttctacag gtgttaacct
      781 agttgctgta cctacaggtt atgttgatac acctaataat acagattttt ccagagttag
      841 tgctaaacca ccgcctggag atcaatttaa acacctcata ccacttatgt acaaaggact
      901 tccttggaat gtagtgcgta taaagattgt acaaatgtta agtgacacac ttaaaaatct
      961 ctctgacaga gtcgtatttg tcttatgggc acatggcttt gagttgacat ctatgaagta
     1021 ttttgtgaaa ataggacctg agcgcacctg ttgtctatgt gatagacgtg ccacatgctt
     1081 ttccactgct tcagacactt atgcctgttg gcatcattct attggatttg attacgtcta
     1141 taatccgttt atgattgatg ttcaacaatg gggttttaca ggtaacctac aaagcaacca
     1201 tgatctgtat tgtcaagtcc atggtaatgc acatgtagct agttgtgatg caatcatgac
     1261 taggtgtcta gctgtccacg agtgctttgt taagcgtgtt gactggacta ttgaatatcc
     1321 tataattggt gatgaactga agattaatgc ggcttgtaga aaggttcaac acatggttgt
     1381 taaagctgca ttattagcag acaaattccc agttcttcac gacattggta accctaaagc
     1441 tattaagtgt gtacctcaag ctgatgtaga atggaagttc tatgatgcac agccttgtag
     1501 tgacaaagct tataaaatag aagaattatt ctattcttat gccacacatt ctgacaaatt
     1561 cacagatggt gtatgcctat tttggaattg caatgtcgat agatatcctg ctaattccat
     1621 tgtttgtaga tttgacacta gagtgctatc taaccttaac ttgcctggtt gtgatggtgg
     1681 cagtttgtat gtaaataaac atgcattcca cacaccagct tttgataaaa gtgcttttgt
     1741 taatttaaaa caattaccat ttttctatta ctctgacagt ccatgtgagt ctcatggaaa
     1801 acaagtagtg tcagatatag attatgtacc actaaagtct gctacgtgta taacacgttg
     1861 caatttaggt ggtgctgtct gtagacatca tgctaatgag tacagattgt atctcgatgc
     1921 ttataacatg atgatctcag ctggctttag cttgtgggtt tacaaacaat ttgatactta
     1981 taacctctgg aacactttta caagacttct cgtgccgaga ggttctaaaa tcgaggaagg
     2041 taagctagtc atttggataa atggagataa gggttacaac ggtttggcag aagtaggcaa
     2101 gaaatttgaa aaggacacag gtatcaaggt gacggttgaa catccagaca agttagaaga
     2161 aaaattccct caagttgctg ctactggtga tggtcccgac attattttct gggcccacga
     2221 tcgttttggt ggatatgctc agtcaggttt gctggctgag atcaccccag acaaggcctt
     2281 ccaagataaa ttgtacccat tcacttggga tgctgtcaga tacaacggta agcttatagc
     2341 atatccaatt gctgttgaag ccttatcgtt gatctacaat aaggacttgt tacctaaccc
     2401 accaaagaca tgggaagaaa ttccagcttt ggataaagag ctaaaggcca agggaaaaag
     2461 cgctttgatg tttaacttac aagaacctta tttcacctgg ccattaatcg ctgctgacgg
     2521 tggttacgca tttaaatacg aaaacggtaa gtatgacatt aaggatgtcg gcgttgataa
     2581 tgctggtgca aaggctggtt tgaccttctt ggttgattta ataaagaata aacacatgaa
     2641 cgctgacact gactactcta tcgccgaagc cgctttcaac aagggtgaaa ctgctatgac
     2701 cataaatggt ccatgggcct ggagtaacat tgatacttct aaagtaaact atggtgtcac
     2761 tgttttgcct acattcaaag gccaaccatc caagcccttt gtcggtgttc tatccgctgg
     2821 catcaatgct gcttctccaa acaaggagct agctaaagaa tttttggaaa attacttgtt
     2881 aactgacgaa ggtttggagg ccgttaacaa ggacaagcca ttgggtgctg tcgctttgaa
     2941 atcttacgaa gaagaattgg ctaaggatcc cagaattgcc gctaccatgg agaacgccca
     3001 aaagggtgaa attatgccaa acatcccaca aatgtcagcc ttctggtacg cagttaggac
     3061 tgccgtgatt aatgctgctt ccggtagaca aaccgtcgac gaagcattaa aagatgctca
     3121 aacgtaaaca ggcccctttt cctttgtcga tatcatgtaa ttagttatgt cacgcttaca
     3181 ttcacgccct cctcccacat ccgctctaac cgaaaaggaa ggagttagac aacctgaagt
     3241 ctaggtccct atttattttt tttaatagtt atgttagtat taagaacgtt atttatattt
     3301 caaatttttc ttttttttct gtacaaagcg gccgccgaac cactagtgca ctgcagtaca
     3361 cggtttcctt gaaatttttt tgattcggta atctccgaac agaaggaaga acgaaggaag
     3421 gagcacagac ttagattggt atatatacgc atatgtagtg ttgaagaaac atgaaattgc
     3481 ccagtattct taacccaact gcacagaaca aaaacgtgca ggaaacgaag ataaatcatg
     3541 tcgaaagcta catataagga acgtgctgct actcatccta gtcctgttgc tgccaagcta
     3601 tttaatatca tgcacgaaaa gcaaacaaac ttgtgtgctt cattggatgt tcgtaccacc
     3661 aaggaattac tggagttagt tgaagcatta ggtcccaaaa tttgtttact aaaaacacat
     3721 gtggatatct tgactgattt ttccatggag ggcacagtta agccgctaaa ggcattatcc
     3781 gccaagtaca attttttact cttcgaggac agaaaatttg ctgacattgg taatacagtc
     3841 aaattgcagt actctgcggg tgtatacaga atagcagaat gggcagacat tacgaatgca
     3901 cacggtgtgg tgggcccagg tattgttagc ggtttgaagc aggcggcaga agaagtaaca
     3961 aaggaaccta gaggcctttt gatgttagca gaattgtcat gcaagggctc cctatctact
     4021 ggagaatata ctaagggtac tgttgacatt gcgaagagcg acaaagattt tgttatcggc
     4081 tttattgctc aaagagacat gggtggaaga gatgaaggtt acgattggtt gattatgaca
     4141 cccggtgtgg gtttagatga caagggagat gcattgggtc aacagtatag aaccgtggat
     4201 gatgtggttt ctacaggatc tgacattatt attgttggaa gaggactatt tgcaaaggga
     4261 agggatgcta aggtagaggg tgaacgttac agaaaagcag gctgggaagc atatttgaga
     4321 agatgcggcc agcaaaacta aaaaactgta ttataagtaa atgcatgtat actaaactca
     4381 caaattagag cttcaattta attatatcag ttattacccg agtaacgaag catctgtgct
     4441 tcattttgta gaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag
     4501 ctgcattttt acagaacaga aatgcaacgc gaaagcgcta ttttaccaac gaagaatctg
     4561 tgcttcattt ttgtaaaaca aaaatgcaac gcgagagcgc taatttttca aacaaagaat
     4621 ctgagctgca tttttacaga acagaaatgc aacgcgagag cgctatttta ccaacaaaga
     4681 atctatactt cttttttgtt ctacaaaaat gcatcccgag agcgctattt ttctaacaaa
     4741 gcatcttaga ttactttttt tctcctttgt gcgctctata atgcagtctc ttgataactt
     4801 tttgcactgt aggtccgtta aggttagaag aaggctactt tggtgtctat tttctcttcc
     4861 ataaaaaaag cctgactcca cttcccgcgt ttactgatta ctagcgaagc tgcgggtgca
     4921 ttttttcaag ataaaggcat ccccgattat attctatacc gatgtggatt gcgcatactt
     4981 tgtgaacaga aagtgatagc gttgatgatt cttcattggt cagaaaatta tgaacggttt
     5041 cttctatttt gtctctatat actacgtata ggaaatgttt acattttcgt attgttttcg
     5101 attcactcta tgaatagttc ttactacaat ttttttgtct aaagagtaat actagagata
     5161 aacataaaaa atgtagaggt cgagtttaga tgcaagttca aggagcgaaa ggtggatggg
     5221 taggttatat agggatatag cacagagata tatagcaaag agatactttt gagcaatgtt
     5281 tgtggaagcg gtattcgcaa tattttagta gctcgttaca gtccggtgcg tttttggttt
     5341 tttgaaagtg cgtcatcaga gcgcttttgg ttttcaaaag cgctctgaag ttcctatact
     5401 ttctagctag agaataggaa cttcccgagc ggccgcgtgt tacaagcttg actccagcgt
     5461 aactggactg caatcaactc actggctcac cttcacgggt gggcctttct tcggtagaaa
     5521 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     5581 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     5641 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     5701 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     5761 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     5821 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     5881 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     5941 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6001 agcgcacgag ggagcttcca ggggtaaacg cctggtatct ttatagtcct gtcgggtttc
     6061 gccacctctg acttgagcat cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6121 aaaacgccag caacgcagaa aggcccaccc gaaggtgagc caggtgatta catttgggcc
     6181 ctcatcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat
     6241 cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga
     6301 gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt
     6361 tttcctgttt ggtcacttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6421 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atgcgggagg
     6481 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     6541 atttatcagc aataaaccag ccagccggga gggccgagcg cagaagtgat cctgcaactt
     6601 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     6661 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     6721 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     6781 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgccaga agtaagttgg
     6841 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     6901 ccgtgagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     6961 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     7021 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgtaaactc tcaaggatct
     7081 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     7141 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     7201 agggaataag ggcgacacgg aaatgttgaa tactcatttt agcttcctta gctcctgaaa
     7261 atctcgataa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg
     7321 aacctcttac gtgccgatca agtcaaaagc ctccggtcgg aggcttttga ctttctgcta
     7381 tggaggtcag gtatgattta aatggtcagt taga
//
