LOCUS       sequence-300431-        6340 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 6340)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6340)
  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-m4046226
FEATURES             Location/Qualifiers
     source          1..6340
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(348..1151)
                     /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"
     primer_bind     1502..1520
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(1565..1588)
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             complement(1589..1606)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     promoter        complement(3198..3611)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      3657..3688
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        3689..3791
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(3792..3860,3861..4652)
                     /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     complement(4010..4029)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             4653..4844
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      4868..4895
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(4907..5494)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5516..5545)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      join(5623..6340,1..265)
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 gtatagattc tttgttggta aaatagcgct ctcgcgttgc atttctgttc tgtaaaaatg
       61 cagctcagat tctttgtttg aaaaattagc gctctcgcgt tgcatttttg ttttacaaaa
      121 atgaagcaca gattcttcgt tggtaaaata gcgctttcgc gttgcatttc tgttctgtaa
      181 aaatgcagct cagattcttt gtttgaaaaa ttagcgctct cgcgttgcat ttttgttcta
      241 caaaatgaag cacagatgct tcgttactcg ggtaataact gatataatta aattgaagct
      301 ctaatttgtg agtttagtat acatgcattt acttataata cagtttttta gttttgctgg
      361 ccgcatcttc tcaaatatgc ttcccagcct gcttttctgt aacgttcacc ctctacctta
      421 gcatcccttc cctttgcaaa tagtcctctt ccaacaataa taatgtcaga tcctgtagaa
      481 accacatcat ccacggttct atactgttga cccaatgcat ctcccttgtc atctaaaccc
      541 acaccgggtg tcataatcaa ccaatcgtaa ccttcatctc ttccacccat gtctctttga
      601 gcaataaagc cgataacaaa atctttgtcg ctcttcgcaa tgtcaacagt acccttagta
      661 tattctccag tagataggga gcccttgcat gacaattctg ctaacatcaa aaggcctcta
      721 ggttcctttg ttacttcttc tgccgcctgc ttcaaaccgc taacaatacc tgggcccacc
      781 acaccgtgtg cattcgtaat gtctgcccat tctgctattc tgtatacacc cgcagagtac
      841 tgcaatttga ctgtattacc aatgtcagca aattttctgt cctcgaagag taaaaaattg
      901 tacttggcgg ataatgcctt tagcggctta actgtgccct ccatggaaaa atcagtcaag
      961 atatccacat gtgtttttag taaacaaatt ttgggaccta atgcttcaac taactccagt
     1021 aattccttgg tggtacgaac atccaatgaa gcacacaagt ttgtttgctt ttcgtgcatg
     1081 atattaaata gcttggcagc aacaggacta ggatgagtag cagcacgttc cttatatgta
     1141 gctttcgaca tgatttatct tcgtttcctg cacgtttttg ttctgtgcag ttgggttaag
     1201 aatactgggc aatttcatgt ttcttcaaca ctacatatgc gtatatatac caatctaagt
     1261 ctgtgctcct tccttcgttc ttccttctgt tcggagatta ccgaatcaaa aaaatttcaa
     1321 ggaaaccgtg tactgcagtg cactagtggt tcggcggccg ctttgtacag aaaaaaaaga
     1381 aaaatttgaa atataaataa cgttcttaat actaacataa ctattaaaaa aaataaatag
     1441 ggacctagac ttcaggttgt ctaactcctt ccttttcggt tagagcggat gtgggaggag
     1501 ggcgtgaatg taagcgtgac ataactaatt acatgatatc gacaaaggaa aaggggcctg
     1561 tttatttttc aaactgcgga tggctccaag aacctctcgg cacgagaagt cttgtaaaag
     1621 tgttccagag gttataagta tcaaattgtt tgtaaaccca caagctaaag ccagctgaga
     1681 tcatcatgtt ataagcatcg agatacaatc tgtactcatt agcatgatgt ctacagacag
     1741 caccacctaa attgcaacgt gttatacacg tagcagactt tagtggtaca taatctatat
     1801 ctgacactac ttgttttcca tgagactcac atggactgtc agagtaatag aaaaatggta
     1861 attgttttaa attaacaaaa gcacttttat caaaagctgg tgtgtggaat gcatgtttat
     1921 ttacatacaa actgccacca tcacaaccag gcaagttaag gttagatagc actctagtgt
     1981 caaatctaca aacaatggaa ttagcaggat atctatcgac attgcaattc caaaataggc
     2041 atacaccatc tgtgaatttg tcagaatgtg tggcataaga atagaataat tcttctattt
     2101 tataagcttt gtcactacaa ggctgtgcat catagaactt ccattctaca tcagcttgag
     2161 gtacacactt aatagcttta gggttaccaa tgtcgtgaag aactgggaat ttgtctgcta
     2221 ataatgcagc tttaacaacc atgtgttgaa cctttctaca agccgcatta atcttcagtt
     2281 catcaccaat tataggatat tcaatagtcc agtcaacacg cttaacaaag cactcgtgga
     2341 cagctagaca cctagtcatg attgcatcac aactagctac atgtgcatta ccatggactt
     2401 gacaatacag atcatggttg ctttgtaggt tacctgtaaa accccattgt tgaacatcaa
     2461 tcataaacgg attatagacg taatcaaatc caatagaatg atgccaacag gcataagtgt
     2521 ctgaagcagt ggaaaagcat gtggcacgtc tatcacatag acaacaggtg cgctcaggtc
     2581 ctattttcac aaaatacttc atagatgtca actcaaagcc atgtgcccat aagacaaata
     2641 cgactctgtc agagagattt ttaagtgtgt cacttaacat ttgtacaatc tttatacgca
     2701 ctacattcca aggaagtcct ttgtacataa gtggtatgag gtgtttaaat tgatctccag
     2761 gcggtggttt agcactaact ctggaaaaat ctgtattatt aggtgtatca acataacctg
     2821 taggtacagc aactaggtta acacctgtag aaaaacctag ctgtaaaggt aaattggtac
     2881 caacagcttc tctagtagca tgacacccct cgacatcgaa gccaatccat gcacgtacat
     2941 gtcttatagc ttcttcgcgg gtgataaaca tgttagggta accattaact tgataattca
     3001 ttttaaaacc catcatagag atgagtcttc tataggtcat gtccttaggt atgccaggta
     3061 tgtcaacaca taaaccttca gttttgaatt tagtgtcaac actgaggtgt gtaggtgcct
     3121 gtgtaggatg taacccagtg attaccttac tacaatcttt aaagagtcct gttacatttt
     3181 ccattttgta attaaaactt agattagatt gctatgcttt ctttctaatg agcaagaagt
     3241 aaaaaaagtt gtaatagaac aagaaaaatg aaactgaaac ttgagaaatt gaagaccgtt
     3301 tattaactta aatatcaatg ggaggtcatc gaaagagaaa aaaatcaaaa aaaaaaattt
     3361 tcaagaaaaa gaaacgtgat aaaaattttt attgcctttt tcgacgaaga aaaagaaacg
     3421 aggcggtctc ttttttcttt tccaaacctt tagtacgggt aattaacgac accctagagg
     3481 aagaaagagg ggaaatttag tatgctgtgc ttgggtgttt tgaagtggta cggcgatgcg
     3541 cggagtccga gaaaatctgg aagagtaaaa aaggagtaga aacattttga agctatggtg
     3601 tgtggggcgg ccgctctaac tgaccattta aatcatacct gacctccata gcagaaagtc
     3661 aaaagcctcc gaccggaggc ttttgacttg atcggcacgt aagaggttcc aactttcacc
     3721 ataatgaaat aagatcacta ccgggcgtat tttttgagtt atcgagattt tcaggagcta
     3781 aggaagctaa aatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     3841 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     3901 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     3961 gtttacgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     4021 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     4081 agaatgactt ggttgagtac tcaccagtca cagaaaagca tctcacggat ggcatgacag
     4141 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     4201 tggcaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     4261 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     4321 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     4381 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggat
     4441 cacttctgcg ctcggccctc ccggctggct ggtttattgc tgataaatct ggagccggtg
     4501 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgcatcg
     4561 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     4621 agataggtgc ctcactgatt aagcattggt aagtgaccaa acaggaaaaa accgccctta
     4681 acatggcccg ctttatcaga agccagacat taacgcttct ggagaaactc aacgagctgg
     4741 acgcggatga acaggcagac atctgtgaat cgcttcacga ccacgctgat gagctttacc
     4801 gcagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgatgaggg cccaaatgta
     4861 atcacctggc tcaccttcgg gtgggccttt ctgcgttgct ggcgtttttc cataggctcc
     4921 gcccccctga cgagcatcac aaaaatcgat gctcaagtca gaggtggcga aacccgacag
     4981 gactataaag ataccaggcg tttacccctg gaagctccct cgtgcgctct cctgttccga
     5041 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     5101 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5161 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     5221 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5281 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5341 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttacctcg gaaaaagagt
     5401 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5461 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgattt tctaccgaag
     5521 aaaggcccac ccgtgaaggt gagccagtga gttgattgca gtccagttac gctggagtca
     5581 agcttgtaac acgcggccgc tcgggaagtt cctattctct agctagaaag tataggaact
     5641 tcagagcgct tttgaaaacc aaaagcgctc tgatgacgca ctttcaaaaa accaaaaacg
     5701 caccggactg taacgagcta ctaaaatatt gcgaataccg cttccacaaa cattgctcaa
     5761 aagtatctct ttgctatata tctctgtgct atatccctat ataacctacc catccacctt
     5821 tcgctccttg aacttgcatc taaactcgac ctctacattt tttatgttta tctctagtat
     5881 tactctttag acaaaaaaat tgtagtaaga actattcata gagtgaatcg aaaacaatac
     5941 gaaaatgtaa acatttccta tacgtagtat atagagacaa aatagaagaa accgttcata
     6001 attttctgac caatgaagaa tcatcaacgc tatcactttc tgttcacaaa gtatgcgcaa
     6061 tccacatcgg tatagaatat aatcggggat gcctttatct tgaaaaaatg cacccgcagc
     6121 ttcgctagta atcagtaaac gcgggaagtg gagtcaggct ttttttatgg aagagaaaat
     6181 agacaccaaa gtagccttct tctaacctta acggacctac agtgcaaaaa gttatcaaga
     6241 gactgcatta tagagcgcac aaaggagaaa aaaagtaatc taagatgctt tgttagaaaa
     6301 atagcgctct cgggatgcat ttttgtagaa caaaaaagaa
//
