LOCUS       sequence-296270-        6435 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 6435)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6435)
  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-m4046587
FEATURES             Location/Qualifiers
     source          1..6435
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        62..503
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    62..179
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     474..497
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             2099..2116
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2117..2182
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(2227..2245)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2596..3399
                     /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      3482..4464
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4542..4571
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4593..5180
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5192..5219)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5243..5434)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5435..6226,6227..6295))
                     /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     6058..6077
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6296..6398)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6399..6430)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 ctatggaggt caggtatgat ttaaatggtc agttagagcg gccgcttgct atattgaagt
       61 acggattaga agccgccgag cgggcgacag ccctccgacg gaagactctc ctccgtgcgt
      121 cctcgtcttc accggtcgcg ttcctgaaac gcagatgtgc ctcgcgccgc actgctccga
      181 acaataaaga ttctacaata ctagctttta tggttatgaa gaggaaaaat tggcagtaac
      241 ctggccccac aaaccttcaa attaacgaat caaattaaca accataggat gataatgcga
      301 ttagtttttt agccttattt ctggggtaat taatcagcga agcgatgatt tttgatctat
      361 taacagatat ataaatggaa aagctgcata accactttaa ctaatacttt caacattttc
      421 agtttgtatt acttcttatt caaatgtcat aaaagtatca acaaaaaatt gttaatatac
      481 ctctatactt taacgtcaag gagaaaaaac tataatggcc gagaatgtca caggcctatt
      541 caaggattgc tcgaaagtga tcaccggttt acaccctacg caggctccaa ctcatttgtc
      601 cgttgacacc aagtttaaga ctgaaggatt gtgtgttgac ataccaggta ttcccaaaga
      661 tatgacttac cgtaggctga tttctatgat gggtttcaag atgaactatc aagttaacgg
      721 ttacccaaat atgtttatca ccagagaaga agctattaga cacgtacgtg catggatcgg
      781 tttcgatgtc gaaggttgtc atgctactag agaagcggtc ggtacaaact tgccattaca
      841 acttggcttt tcaaccggtg ttaatttggt tgctgttcct acagggtatg tcgacactcc
      901 aaacaacacg gatttctctc gagtgtccgc caagccacca cctggtgatc aattcaaaca
      961 cttgatcccg ctcatgtaca agggtttgcc ttggaacgtt gtcagaatta aaatagtaca
     1021 aatgttatct gatactttga agaatctgag tgacagagtt gtcttcgtcc tttgggctca
     1081 cggttttgag ctaaccagca tgaagtactt tgtgaagatt ggaccagaac ggacttgttg
     1141 cttgtgtgac agaagagcca cttgtttctc caccgctagc gatacatatg catgttggca
     1201 tcattctatt ggtttcgact acgtttacaa cccctttatg atcgacgttc aacagtgggg
     1261 tttcaccggt aatttgcaat cgaaccacga cttgtattgt caagttcacg gcaatgctca
     1321 cgtggcttca tgcgatgcta ttatgacaag gtgtttagcc gtccatgaat gtttcgttaa
     1381 aagagtagac tggactatcg aatacccaat tatcggtgat gaactcaaga ttaacgctgc
     1441 ctgtagaaag gtccaacata tggttgtaaa ggcagctcta ttagctgaca aatttccagt
     1501 tttgcacgat atagggaatc caaaggccat aaagtgtgtt ccacaagctg acgtcgaatg
     1561 gaaattctac gatgcgcaac cctgctctga taaagcttat aagatcgagg aattgtttta
     1621 ctcttacgct acccactccg ataagttcac tgacggtgtg tgtcttttct ggaactgcaa
     1681 tgttgataga tatccagcaa actccatcgt ttgtagattc gacacgcgag tcttgagcaa
     1741 cttgaactta ccaggttgtg atggtggttc cttgtacgtc aataaacatg catttcacac
     1801 tcccgctttc gacaagtctg cttttgttaa tttaaagcaa ttgccttttt tctattactc
     1861 tgatagtcca tgcgaatctc atggtaagca agtcgtttcc gacattgatt acgtaccact
     1921 gaagtcagcc acctgtataa caagatgtaa ccttggaggt gctgtctgcc gccaccatgc
     1981 taacgagtac agactatact tggatgccta taacatgatg atttcagccg gtttctcttt
     2041 atgggtttac aagcagtttg acacttacaa cttgtggaac accttcacgc gtttgcaact
     2101 cgtgccgaga ggttctgatt ataaagatca tgatggtgat tataaagatc atgatattga
     2161 ttataaagat gatgatgata aataaacagg ccccttttcc tttgtcgata tcatgtaatt
     2221 agttatgtca cgcttacatt cacgccctcc tcccacatcc gctctaaccg aaaaggaagg
     2281 agttagacaa cctgaagtct aggtccctat ttattttttt taatagttat gttagtatta
     2341 agaacgttat ttatatttca aatttttctt ttttttctgt acaaagcggc cgccgaacca
     2401 ctagtgcact gcagtacacg gtttccttga aatttttttg attcggtaat ctccgaacag
     2461 aaggaagaac gaaggaagga gcacagactt agattggtat atatacgcat atgtagtgtt
     2521 gaagaaacat gaaattgccc agtattctta acccaactgc acagaacaaa aacgtgcagg
     2581 aaacgaagat aaatcatgtc gaaagctaca tataaggaac gtgctgctac tcatcctagt
     2641 cctgttgctg ccaagctatt taatatcatg cacgaaaagc aaacaaactt gtgtgcttca
     2701 ttggatgttc gtaccaccaa ggaattactg gagttagttg aagcattagg tcccaaaatt
     2761 tgtttactaa aaacacatgt ggatatcttg actgattttt ccatggaggg cacagttaag
     2821 ccgctaaagg cattatccgc caagtacaat tttttactct tcgaggacag aaaatttgct
     2881 gacattggta atacagtcaa attgcagtac tctgcgggtg tatacagaat agcagaatgg
     2941 gcagacatta cgaatgcaca cggtgtggtg ggcccaggta ttgttagcgg tttgaagcag
     3001 gcggcagaag aagtaacaaa ggaacctaga ggccttttga tgttagcaga attgtcatgc
     3061 aagggctccc tatctactgg agaatatact aagggtactg ttgacattgc gaagagcgac
     3121 aaagattttg ttatcggctt tattgctcaa agagacatgg gtggaagaga tgaaggttac
     3181 gattggttga ttatgacacc cggtgtgggt ttagatgaca agggagatgc attgggtcaa
     3241 cagtatagaa ccgtggatga tgtggtttct acaggatctg acattattat tgttggaaga
     3301 ggactatttg caaagggaag ggatgctaag gtagagggtg aacgttacag aaaagcaggc
     3361 tgggaagcat atttgagaag atgcggccag caaaactaaa aaactgtatt ataagtaaat
     3421 gcatgtatac taaactcaca aattagagct tcaatttaat tatatcagtt attacccgag
     3481 taacgaagca tctgtgcttc attttgtaga acaaaaatgc aacgcgagag cgctaatttt
     3541 tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga aagcgctatt
     3601 ttaccaacga agaatctgtg cttcattttt gtaaaacaaa aatgcaacgc gagagcgcta
     3661 atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgagagcg
     3721 ctattttacc aacaaagaat ctatacttct tttttgttct acaaaaatgc atcccgagag
     3781 cgctattttt ctaacaaagc atcttagatt actttttttc tcctttgtgc gctctataat
     3841 gcagtctctt gataactttt tgcactgtag gtccgttaag gttagaagaa ggctactttg
     3901 gtgtctattt tctcttccat aaaaaaagcc tgactccact tcccgcgttt actgattact
     3961 agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc ccgattatat tctataccga
     4021 tgtggattgc gcatactttg tgaacagaaa gtgatagcgt tgatgattct tcattggtca
     4081 gaaaattatg aacggtttct tctattttgt ctctatatac tacgtatagg aaatgtttac
     4141 attttcgtat tgttttcgat tcactctatg aatagttctt actacaattt ttttgtctaa
     4201 agagtaatac tagagataaa cataaaaaat gtagaggtcg agtttagatg caagttcaag
     4261 gagcgaaagg tggatgggta ggttatatag ggatatagca cagagatata tagcaaagag
     4321 atacttttga gcaatgtttg tggaagcggt attcgcaata ttttagtagc tcgttacagt
     4381 ccggtgcgtt tttggttttt tgaaagtgcg tcatcagagc gcttttggtt ttcaaaagcg
     4441 ctctgaagtt cctatacttt ctagctagag aataggaact tcccgagcgg ccgcgtgtta
     4501 caagcttgac tccagcgtaa ctggactgca atcaactcac tggctcacct tcacgggtgg
     4561 gcctttcttc ggtagaaaat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     4621 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     4681 ctaccaactc tttttccgag gtaactggct tcagcagagc gcagatacca aatactgttc
     4741 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     4801 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     4861 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     4921 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     4981 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     5041 gcagggtcgg aacaggagag cgcacgaggg agcttccagg ggtaaacgcc tggtatcttt
     5101 atagtcctgt cgggtttcgc cacctctgac ttgagcatcg atttttgtga tgctcgtcag
     5161 gggggcggag cctatggaaa aacgccagca acgcagaaag gcccacccga aggtgagcca
     5221 ggtgattaca tttgggccct catcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc
     5281 agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat
     5341 ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg
     5401 ccatgttaag ggcggttttt tcctgtttgg tcacttacca atgcttaatc agtgaggcac
     5461 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     5521 taactacgat gcgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     5581 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccgggagg gccgagcgca
     5641 gaagtgatcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     5701 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     5761 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     5821 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     5881 ttgccagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     5941 ctcttactgt catgccatcc gtgagatgct tttctgtgac tggtgagtac tcaaccaagt
     6001 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     6061 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     6121 gtaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     6181 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     6241 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcattttag
     6301 cttccttagc tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt
     6361 cattatggtg aaagttggaa cctcttacgt gccgatcaag tcaaaagcct ccggtcggag
     6421 gcttttgact ttctg
//
