LOCUS       sequence-297827-        6382 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 6382)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6382)
  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-m4046535
FEATURES             Location/Qualifiers
     source          1..6382
                     /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"
     CDS             2027..2092
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(2137..2155)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2506..3309
                     /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      3392..4374
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4452..4481
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4503..5090
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5102..5129)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5153..5344)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5345..6136,6137..6205))
                     /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     5968..5987
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6206..6308)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6309..6340)
                     /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 ggttctgatt ataaagatca
     2041 tgatggtgat tataaagatc atgatattga ttataaagat gatgatgata aataaacagg
     2101 ccccttttcc tttgtcgata tcatgtaatt agttatgtca cgcttacatt cacgccctcc
     2161 tcccacatcc gctctaaccg aaaaggaagg agttagacaa cctgaagtct aggtccctat
     2221 ttattttttt taatagttat gttagtatta agaacgttat ttatatttca aatttttctt
     2281 ttttttctgt acaaagcggc cgccgaacca ctagtgcact gcagtacacg gtttccttga
     2341 aatttttttg attcggtaat ctccgaacag aaggaagaac gaaggaagga gcacagactt
     2401 agattggtat atatacgcat atgtagtgtt gaagaaacat gaaattgccc agtattctta
     2461 acccaactgc acagaacaaa aacgtgcagg aaacgaagat aaatcatgtc gaaagctaca
     2521 tataaggaac gtgctgctac tcatcctagt cctgttgctg ccaagctatt taatatcatg
     2581 cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc gtaccaccaa ggaattactg
     2641 gagttagttg aagcattagg tcccaaaatt tgtttactaa aaacacatgt ggatatcttg
     2701 actgattttt ccatggaggg cacagttaag ccgctaaagg cattatccgc caagtacaat
     2761 tttttactct tcgaggacag aaaatttgct gacattggta atacagtcaa attgcagtac
     2821 tctgcgggtg tatacagaat agcagaatgg gcagacatta cgaatgcaca cggtgtggtg
     2881 ggcccaggta ttgttagcgg tttgaagcag gcggcagaag aagtaacaaa ggaacctaga
     2941 ggccttttga tgttagcaga attgtcatgc aagggctccc tatctactgg agaatatact
     3001 aagggtactg ttgacattgc gaagagcgac aaagattttg ttatcggctt tattgctcaa
     3061 agagacatgg gtggaagaga tgaaggttac gattggttga ttatgacacc cggtgtgggt
     3121 ttagatgaca agggagatgc attgggtcaa cagtatagaa ccgtggatga tgtggtttct
     3181 acaggatctg acattattat tgttggaaga ggactatttg caaagggaag ggatgctaag
     3241 gtagagggtg aacgttacag aaaagcaggc tgggaagcat atttgagaag atgcggccag
     3301 caaaactaaa aaactgtatt ataagtaaat gcatgtatac taaactcaca aattagagct
     3361 tcaatttaat tatatcagtt attacccgag taacgaagca tctgtgcttc attttgtaga
     3421 acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac
     3481 agaacagaaa tgcaacgcga aagcgctatt ttaccaacga agaatctgtg cttcattttt
     3541 gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt
     3601 tttacagaac agaaatgcaa cgcgagagcg ctattttacc aacaaagaat ctatacttct
     3661 tttttgttct acaaaaatgc atcccgagag cgctattttt ctaacaaagc atcttagatt
     3721 actttttttc tcctttgtgc gctctataat gcagtctctt gataactttt tgcactgtag
     3781 gtccgttaag gttagaagaa ggctactttg gtgtctattt tctcttccat aaaaaaagcc
     3841 tgactccact tcccgcgttt actgattact agcgaagctg cgggtgcatt ttttcaagat
     3901 aaaggcatcc ccgattatat tctataccga tgtggattgc gcatactttg tgaacagaaa
     3961 gtgatagcgt tgatgattct tcattggtca gaaaattatg aacggtttct tctattttgt
     4021 ctctatatac tacgtatagg aaatgtttac attttcgtat tgttttcgat tcactctatg
     4081 aatagttctt actacaattt ttttgtctaa agagtaatac tagagataaa cataaaaaat
     4141 gtagaggtcg agtttagatg caagttcaag gagcgaaagg tggatgggta ggttatatag
     4201 ggatatagca cagagatata tagcaaagag atacttttga gcaatgtttg tggaagcggt
     4261 attcgcaata ttttagtagc tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg
     4321 tcatcagagc gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt ctagctagag
     4381 aataggaact tcccgagcgg ccgcgtgtta caagcttgac tccagcgtaa ctggactgca
     4441 atcaactcac tggctcacct tcacgggtgg gcctttcttc ggtagaaaat caaaggatct
     4501 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4561 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgag gtaactggct
     4621 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     4681 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     4741 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     4801 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     4861 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     4921 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     4981 agcttccagg ggtaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     5041 ttgagcatcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     5101 acgcagaaag gcccacccga aggtgagcca ggtgattaca tttgggccct catcagaggt
     5161 tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt
     5221 gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa
     5281 gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg
     5341 tcacttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     5401 ccatagttgc ctgactcccc gtcgtgtaga taactacgat gcgggagggc ttaccatctg
     5461 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     5521 taaaccagcc agccgggagg gccgagcgca gaagtgatcc tgcaacttta tccgcctcca
     5581 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     5641 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     5701 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     5761 aagcggttag ctccttcggt cctccgatcg ttgccagaag taagttggcc gcagtgttat
     5821 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtgagatgct
     5881 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     5941 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     6001 tgctcatcat tggaaaacgt tcttcggggc gtaaactctc aaggatctta ccgctgttga
     6061 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     6121 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     6181 cgacacggaa atgttgaata ctcattttag cttccttagc tcctgaaaat ctcgataact
     6241 caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt
     6301 gccgatcaag tcaaaagcct ccggtcggag gcttttgact ttctgctatg gaggtcaggt
     6361 atgatttaaa tggtcagtta ga
//
