LOCUS       sequence-298000-        6393 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 6393)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6393)
  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-m4046342
FEATURES             Location/Qualifiers
     source          1..6393
                     /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             2062..2079
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2080..2103
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(2148..2166)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2517..3320
                     /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      3403..4385
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4463..4492
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4514..5101
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5113..5140)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5164..5355)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5356..6147,6148..6216))
                     /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     5979..5998
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6217..6319)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6320..6351)
                     /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 gccgagaatg tcacaggcct attcaaggat tgctcgaaag tgatcaccgg tttacaccct
      541 acgcaggctc caactcattt gtccgttgac accaagttta agactgaagg attgtgtgtt
      601 gacataccag gtattcccaa agatatgact taccgtaggc tgatttctat gatgggtttc
      661 aagatgaact atcaagttaa cggttaccca aatatgttta tcaccagaga agaagctatt
      721 agacacgtac gtgcatggat cggtttcgat gtcgaaggtt gtcatgctac tagagaagcg
      781 gtcggtacaa acttgccatt acaacttggc ttttcaaccg gtgttaattt ggttgctgtt
      841 cctacagggt atgtcgacac tccaaacaac acggatttct ctcgagtgtc cgccaagcca
      901 ccacctggtg atcaattcaa acacttgatc ccgctcatgt acaagggttt gccttggaac
      961 gttgtcagaa ttaaaatagt acaaatgtta tctgatactt tgaagaatct gagtgacaga
     1021 gttgtcttcg tcctttgggc tcacggtttt gagctaacca gcatgaagta ctttgtgaag
     1081 attggaccag aacggacttg ttgcttgtgt gacagaagag ccacttgttt ctccaccgct
     1141 agcgatacat atgcatgttg gcatcattct attggtttcg actacgttta caaccccttt
     1201 atgatcgacg ttcaacagtg gggtttcacc ggtaatttgc aatcgaacca cgacttgtat
     1261 tgtcaagttc acggcaatgc tcacgtggct tcatgcgatg ctattatgac aaggtgttta
     1321 gccgtccatg aatgtttcgt taaaagagta gactggacta tcgaataccc aattatcggt
     1381 gatgaactca agattaacgc tgcctgtaga aaggtccaac atatggttgt aaaggcagct
     1441 ctattagctg acaaatttcc agttttgcac gatataggga atccaaaggc cataaagtgt
     1501 gttccacaag ctgacgtcga atggaaattc tacgatgcgc aaccctgctc tgataaagct
     1561 tataagatcg aggaattgtt ttactcttac gctacccact ccgataagtt cactgacggt
     1621 gtgtgtcttt tctggaactg caatgttgat agatatccag caaactccat cgtttgtaga
     1681 ttcgacacgc gagtcttgag caacttgaac ttaccaggtt gtgatggtgg ttccttgtac
     1741 gtcaataaac atgcatttca cactcccgct ttcgacaagt ctgcttttgt taatttaaag
     1801 caattgcctt ttttctatta ctctgatagt ccatgcgaat ctcatggtaa gcaagtcgtt
     1861 tccgacattg attacgtacc actgaagtca gccacctgta taacaagatg taaccttgga
     1921 ggtgctgtct gccgccacca tgctaacgag tacagactat acttggatgc ctataacatg
     1981 atgatttcag ccggtttctc tttatgggtt tacaagcagt ttgacactta caacttgtgg
     2041 aacaccttca cgcgtttgca actcgtgccg agaggttctt ggagccatcc gcagtttgaa
     2101 aaataaacag gccccttttc ctttgtcgat atcatgtaat tagttatgtc acgcttacat
     2161 tcacgccctc ctcccacatc cgctctaacc gaaaaggaag gagttagaca acctgaagtc
     2221 taggtcccta tttatttttt ttaatagtta tgttagtatt aagaacgtta tttatatttc
     2281 aaatttttct tttttttctg tacaaagcgg ccgccgaacc actagtgcac tgcagtacac
     2341 ggtttccttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg
     2401 agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc
     2461 cagtattctt aacccaactg cacagaacaa aaacgtgcag gaaacgaaga taaatcatgt
     2521 cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct gccaagctat
     2581 ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt cgtaccacca
     2641 aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta aaaacacatg
     2701 tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag gcattatccg
     2761 ccaagtacaa ttttttactc ttcgaggaca gaaaatttgc tgacattggt aatacagtca
     2821 aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt acgaatgcac
     2881 acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa gaagtaacaa
     2941 aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc ctatctactg
     3001 gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt gttatcggct
     3061 ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg attatgacac
     3121 ccggtgtggg tttagatgac aagggagatg cattgggtca acagtataga accgtggatg
     3181 atgtggtttc tacaggatct gacattatta ttgttggaag aggactattt gcaaagggaa
     3241 gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca tatttgagaa
     3301 gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata ctaaactcac
     3361 aaattagagc ttcaatttaa ttatatcagt tattacccga gtaacgaagc atctgtgctt
     3421 cattttgtag aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc
     3481 tgcattttta cagaacagaa atgcaacgcg aaagcgctat tttaccaacg aagaatctgt
     3541 gcttcatttt tgtaaaacaa aaatgcaacg cgagagcgct aatttttcaa acaaagaatc
     3601 tgagctgcat ttttacagaa cagaaatgca acgcgagagc gctattttac caacaaagaa
     3661 tctatacttc ttttttgttc tacaaaaatg catcccgaga gcgctatttt tctaacaaag
     3721 catcttagat tacttttttt ctcctttgtg cgctctataa tgcagtctct tgataacttt
     3781 ttgcactgta ggtccgttaa ggttagaaga aggctacttt ggtgtctatt ttctcttcca
     3841 taaaaaaagc ctgactccac ttcccgcgtt tactgattac tagcgaagct gcgggtgcat
     3901 tttttcaaga taaaggcatc cccgattata ttctataccg atgtggattg cgcatacttt
     3961 gtgaacagaa agtgatagcg ttgatgattc ttcattggtc agaaaattat gaacggtttc
     4021 ttctattttg tctctatata ctacgtatag gaaatgttta cattttcgta ttgttttcga
     4081 ttcactctat gaatagttct tactacaatt tttttgtcta aagagtaata ctagagataa
     4141 acataaaaaa tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag gtggatgggt
     4201 aggttatata gggatatagc acagagatat atagcaaaga gatacttttg agcaatgttt
     4261 gtggaagcgg tattcgcaat attttagtag ctcgttacag tccggtgcgt ttttggtttt
     4321 ttgaaagtgc gtcatcagag cgcttttggt tttcaaaagc gctctgaagt tcctatactt
     4381 tctagctaga gaataggaac ttcccgagcg gccgcgtgtt acaagcttga ctccagcgta
     4441 actggactgc aatcaactca ctggctcacc ttcacgggtg ggcctttctt cggtagaaaa
     4501 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     4561 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     4621 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     4681 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     4741 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     4801 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     4861 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     4921 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     4981 gcgcacgagg gagcttccag gggtaaacgc ctggtatctt tatagtcctg tcgggtttcg
     5041 ccacctctga cttgagcatc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     5101 aaacgccagc aacgcagaaa ggcccacccg aaggtgagcc aggtgattac atttgggccc
     5161 tcatcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc
     5221 agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag
     5281 tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt
     5341 ttcctgtttg gtcacttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5401 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tgcgggaggg
     5461 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5521 tttatcagca ataaaccagc cagccgggag ggccgagcgc agaagtgatc ctgcaacttt
     5581 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5641 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5701 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5761 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgccagaa gtaagttggc
     5821 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5881 cgtgagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5941 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6001 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgtaaactct caaggatctt
     6061 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6121 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6181 gggaataagg gcgacacgga aatgttgaat actcatttta gcttccttag ctcctgaaaa
     6241 tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt gaaagttgga
     6301 acctcttacg tgccgatcaa gtcaaaagcc tccggtcgga ggcttttgac tttctgctat
     6361 ggaggtcagg tatgatttaa atggtcagtt aga
//
