LOCUS       sequence-291545-        6429 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 6429)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6429)
  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-m4046579
FEATURES             Location/Qualifiers
     source          1..6429
                     /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..2139
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(2184..2202)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2553..3356
                     /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      3439..4421
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4499..4528
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4550..5137
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5149..5176)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5200..5391)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5392..6183,6184..6252))
                     /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     6015..6034
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6253..6355)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6356..6387)
                     /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 tctgattata aagatcatga tggtgattat
     2101 aaagatcatg atattgatta taaagatgat gatgataaat aaacaggccc cttttccttt
     2161 gtcgatatca tgtaattagt tatgtcacgc ttacattcac gccctcctcc cacatccgct
     2221 ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta ttttttttaa
     2281 tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt tttctgtaca
     2341 aagcggccgc cgaaccacta gtgcactgca gtacacggtt tccttgaaat ttttttgatt
     2401 cggtaatctc cgaacagaag gaagaacgaa ggaaggagca cagacttaga ttggtatata
     2461 tacgcatatg tagtgttgaa gaaacatgaa attgcccagt attcttaacc caactgcaca
     2521 gaacaaaaac gtgcaggaaa cgaagataaa tcatgtcgaa agctacatat aaggaacgtg
     2581 ctgctactca tcctagtcct gttgctgcca agctatttaa tatcatgcac gaaaagcaaa
     2641 caaacttgtg tgcttcattg gatgttcgta ccaccaagga attactggag ttagttgaag
     2701 cattaggtcc caaaatttgt ttactaaaaa cacatgtgga tatcttgact gatttttcca
     2761 tggagggcac agttaagccg ctaaaggcat tatccgccaa gtacaatttt ttactcttcg
     2821 aggacagaaa atttgctgac attggtaata cagtcaaatt gcagtactct gcgggtgtat
     2881 acagaatagc agaatgggca gacattacga atgcacacgg tgtggtgggc ccaggtattg
     2941 ttagcggttt gaagcaggcg gcagaagaag taacaaagga acctagaggc cttttgatgt
     3001 tagcagaatt gtcatgcaag ggctccctat ctactggaga atatactaag ggtactgttg
     3061 acattgcgaa gagcgacaaa gattttgtta tcggctttat tgctcaaaga gacatgggtg
     3121 gaagagatga aggttacgat tggttgatta tgacacccgg tgtgggttta gatgacaagg
     3181 gagatgcatt gggtcaacag tatagaaccg tggatgatgt ggtttctaca ggatctgaca
     3241 ttattattgt tggaagagga ctatttgcaa agggaaggga tgctaaggta gagggtgaac
     3301 gttacagaaa agcaggctgg gaagcatatt tgagaagatg cggccagcaa aactaaaaaa
     3361 ctgtattata agtaaatgca tgtatactaa actcacaaat tagagcttca atttaattat
     3421 atcagttatt acccgagtaa cgaagcatct gtgcttcatt ttgtagaaca aaaatgcaac
     3481 gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc
     3541 aacgcgaaag cgctatttta ccaacgaaga atctgtgctt catttttgta aaacaaaaat
     3601 gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga
     3661 aatgcaacgc gagagcgcta ttttaccaac aaagaatcta tacttctttt ttgttctaca
     3721 aaaatgcatc ccgagagcgc tatttttcta acaaagcatc ttagattact ttttttctcc
     3781 tttgtgcgct ctataatgca gtctcttgat aactttttgc actgtaggtc cgttaaggtt
     3841 agaagaaggc tactttggtg tctattttct cttccataaa aaaagcctga ctccacttcc
     3901 cgcgtttact gattactagc gaagctgcgg gtgcattttt tcaagataaa ggcatccccg
     3961 attatattct ataccgatgt ggattgcgca tactttgtga acagaaagtg atagcgttga
     4021 tgattcttca ttggtcagaa aattatgaac ggtttcttct attttgtctc tatatactac
     4081 gtataggaaa tgtttacatt ttcgtattgt tttcgattca ctctatgaat agttcttact
     4141 acaatttttt tgtctaaaga gtaatactag agataaacat aaaaaatgta gaggtcgagt
     4201 ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt tatataggga tatagcacag
     4261 agatatatag caaagagata cttttgagca atgtttgtgg aagcggtatt cgcaatattt
     4321 tagtagctcg ttacagtccg gtgcgttttt ggttttttga aagtgcgtca tcagagcgct
     4381 tttggttttc aaaagcgctc tgaagttcct atactttcta gctagagaat aggaacttcc
     4441 cgagcggccg cgtgttacaa gcttgactcc agcgtaactg gactgcaatc aactcactgg
     4501 ctcaccttca cgggtgggcc tttcttcggt agaaaatcaa aggatcttct tgagatcctt
     4561 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     4621 gtttgccgga tcaagagcta ccaactcttt ttccgaggta actggcttca gcagagcgca
     4681 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4741 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4801 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4861 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4921 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4981 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggt
     5041 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcatcgatt
     5101 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cagaaaggcc
     5161 cacccgaagg tgagccaggt gattacattt gggccctcat cagaggtttt caccgtcatc
     5221 accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca
     5281 gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg
     5341 gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca cttaccaatg
     5401 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     5461 actccccgtc gtgtagataa ctacgatgcg ggagggctta ccatctggcc ccagtgctgc
     5521 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     5581 cgggagggcc gagcgcagaa gtgatcctgc aactttatcc gcctccatcc agtctattaa
     5641 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     5701 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     5761 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     5821 cttcggtcct ccgatcgttg ccagaagtaa gttggccgca gtgttatcac tcatggttat
     5881 ggcagcactg cataattctc ttactgtcat gccatccgtg agatgctttt ctgtgactgg
     5941 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6001 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6061 aaaacgttct tcggggcgta aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6121 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6181 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6241 ttgaatactc attttagctt ccttagctcc tgaaaatctc gataactcaa aaaatacgcc
     6301 cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc gatcaagtca
     6361 aaagcctccg gtcggaggct tttgactttc tgctatggag gtcaggtatg atttaaatgg
     6421 tcagttaga
//
