LOCUS       sequence-307359-        6345 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 6345)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6345)
  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-m4046657
FEATURES             Location/Qualifiers
     source          1..6345
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        40..481
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    40..157
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     452..475
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     complement(2115..2133)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2484..3287
                     /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      3370..4352
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4430..4459
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4481..5068
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5080..5107)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5131..5322)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5323..6114,6115..6183))
                     /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     5946..5965
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6184..6286)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6287..6318)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 aaatggtcag ttagagcggc cgcttgctat attgaagtac ggattagaag ccgccgagcg
       61 ggcgacagcc ctccgacgga agactctcct ccgtgcgtcc tcgtcttcac cggtcgcgtt
      121 cctgaaacgc agatgtgcct cgcgccgcac tgctccgaac aataaagatt ctacaatact
      181 agcttttatg gttatgaaga ggaaaaattg gcagtaacct ggccccacaa accttcaaat
      241 taacgaatca aattaacaac cataggatga taatgcgatt agttttttag ccttatttct
      301 ggggtaatta atcagcgaag cgatgatttt tgatctatta acagatatat aaatggaaaa
      361 gctgcataac cactttaact aatactttca acattttcag tttgtattac ttcttattca
      421 aatgtcataa aagtatcaac aaaaaattgt taatatacct ctatacttta acgtcaagga
      481 gaaaaaacta taatggaaaa tgtaacagga ctctttaaag attgtagtaa ggtaatcact
      541 gggttacatc ctacacaggc acctacacac ctcagtgttg acactaaatt caaaactgaa
      601 ggtttatgtg ttgacatacc tggcatacct aaggacatga cctatagaag actcatctct
      661 atgatgggtt ttaaaatgaa ttatcaagtt aatggttacc ctaacatgtt tatcacccgc
      721 gaagaagcta taagacatgt acgtgcatgg attggcttcg atgtcgaggg gtgtcatgct
      781 actagagaag ctgttggtac caatttacct ttacagctag gtttttctac aggtgttaac
      841 ctagttgctg tacctacagg ttatgttgat acacctaata atacagattt ttccagagtt
      901 agtgctaaac caccgcctgg agatcaattt aaacacctca taccacttat gtacaaagga
      961 cttccttgga atgtagtgcg tataaagatt gtacaaatgt taagtgacac acttaaaaat
     1021 ctctctgaca gagtcgtatt tgtcttatgg gcacatggct ttgagttgac atctatgaag
     1081 tattttgtga aaataggacc tgagcgcacc tgttgtctat gtgatagacg tgccacatgc
     1141 ttttccactg cttcagacac ttatgcctgt tggcatcatt ctattggatt tgattacgtc
     1201 tataatccgt ttatgattga tgttcaacaa tggggtttta caggtaacct acaaagcaac
     1261 catgatctgt attgtcaagt ccatggtaat gcacatgtag ctagttgtga tgcaatcatg
     1321 actaggtgtc tagctgtcca cgagtgcttt gttaagcgtg ttgactggac tattgaatat
     1381 cctataattg gtgatgaact gaagattaat gcggcttgta gaaaggttca acacatggtt
     1441 gttaaagctg cattattagc agacaaattc ccagttcttc acgacattgg taaccctaaa
     1501 gctattaagt gtgtacctca agctgatgta gaatggaagt tctatgatgc acagccttgt
     1561 agtgacaaag cttataaaat agaagaatta ttctattctt atgccacaca ttctgacaaa
     1621 ttcacagatg gtgtatgcct attttggaat tgcaatgtcg atagatatcc tgctaattcc
     1681 attgtttgta gatttgacac tagagtgcta tctaacctta acttgcctgg ttgtgatggt
     1741 ggcagtttgt atgtaaataa acatgcattc cacacaccag cttttgataa aagtgctttt
     1801 gttaatttaa aacaattacc atttttctat tactctgaca gtccatgtga gtctcatgga
     1861 aaacaagtag tgtcagatat agattatgta ccactaaagt ctgctacgtg tataacacgt
     1921 tgcaatttag gtggtgctgt ctgtagacat catgctaatg agtacagatt gtatctcgat
     1981 gcttataaca tgatgatctc agctggcttt agcttgtggg tttacaaaca atttgatact
     2041 tataacctct ggaacacttt tacaagactt taaacaggcc ccttttcctt tgtcgatatc
     2101 atgtaattag ttatgtcacg cttacattca cgccctcctc ccacatccgc tctaaccgaa
     2161 aaggaaggag ttagacaacc tgaagtctag gtccctattt atttttttta atagttatgt
     2221 tagtattaag aacgttattt atatttcaaa tttttctttt ttttctgtac aaagcggccg
     2281 ccgaaccact agtgcactgc agtacacggt ttccttgaaa tttttttgat tcggtaatct
     2341 ccgaacagaa ggaagaacga aggaaggagc acagacttag attggtatat atacgcatat
     2401 gtagtgttga agaaacatga aattgcccag tattcttaac ccaactgcac agaacaaaaa
     2461 cgtgcaggaa acgaagataa atcatgtcga aagctacata taaggaacgt gctgctactc
     2521 atcctagtcc tgttgctgcc aagctattta atatcatgca cgaaaagcaa acaaacttgt
     2581 gtgcttcatt ggatgttcgt accaccaagg aattactgga gttagttgaa gcattaggtc
     2641 ccaaaatttg tttactaaaa acacatgtgg atatcttgac tgatttttcc atggagggca
     2701 cagttaagcc gctaaaggca ttatccgcca agtacaattt tttactcttc gaggacagaa
     2761 aatttgctga cattggtaat acagtcaaat tgcagtactc tgcgggtgta tacagaatag
     2821 cagaatgggc agacattacg aatgcacacg gtgtggtggg cccaggtatt gttagcggtt
     2881 tgaagcaggc ggcagaagaa gtaacaaagg aacctagagg ccttttgatg ttagcagaat
     2941 tgtcatgcaa gggctcccta tctactggag aatatactaa gggtactgtt gacattgcga
     3001 agagcgacaa agattttgtt atcggcttta ttgctcaaag agacatgggt ggaagagatg
     3061 aaggttacga ttggttgatt atgacacccg gtgtgggttt agatgacaag ggagatgcat
     3121 tgggtcaaca gtatagaacc gtggatgatg tggtttctac aggatctgac attattattg
     3181 ttggaagagg actatttgca aagggaaggg atgctaaggt agagggtgaa cgttacagaa
     3241 aagcaggctg ggaagcatat ttgagaagat gcggccagca aaactaaaaa actgtattat
     3301 aagtaaatgc atgtatacta aactcacaaa ttagagcttc aatttaatta tatcagttat
     3361 tacccgagta acgaagcatc tgtgcttcat tttgtagaac aaaaatgcaa cgcgagagcg
     3421 ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg caacgcgaaa
     3481 gcgctatttt accaacgaag aatctgtgct tcatttttgt aaaacaaaaa tgcaacgcga
     3541 gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg
     3601 cgagagcgct attttaccaa caaagaatct atacttcttt tttgttctac aaaaatgcat
     3661 cccgagagcg ctatttttct aacaaagcat cttagattac tttttttctc ctttgtgcgc
     3721 tctataatgc agtctcttga taactttttg cactgtaggt ccgttaaggt tagaagaagg
     3781 ctactttggt gtctattttc tcttccataa aaaaagcctg actccacttc ccgcgtttac
     3841 tgattactag cgaagctgcg ggtgcatttt ttcaagataa aggcatcccc gattatattc
     3901 tataccgatg tggattgcgc atactttgtg aacagaaagt gatagcgttg atgattcttc
     3961 attggtcaga aaattatgaa cggtttcttc tattttgtct ctatatacta cgtataggaa
     4021 atgtttacat tttcgtattg ttttcgattc actctatgaa tagttcttac tacaattttt
     4081 ttgtctaaag agtaatacta gagataaaca taaaaaatgt agaggtcgag tttagatgca
     4141 agttcaagga gcgaaaggtg gatgggtagg ttatataggg atatagcaca gagatatata
     4201 gcaaagagat acttttgagc aatgtttgtg gaagcggtat tcgcaatatt ttagtagctc
     4261 gttacagtcc ggtgcgtttt tggttttttg aaagtgcgtc atcagagcgc ttttggtttt
     4321 caaaagcgct ctgaagttcc tatactttct agctagagaa taggaacttc ccgagcggcc
     4381 gcgtgttaca agcttgactc cagcgtaact ggactgcaat caactcactg gctcaccttc
     4441 acgggtgggc ctttcttcgg tagaaaatca aaggatcttc ttgagatcct ttttttctgc
     4501 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     4561 atcaagagct accaactctt tttccgaggt aactggcttc agcagagcgc agataccaaa
     4621 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4681 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4741 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4801 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     4861 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4921 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg taaacgcctg
     4981 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcatcgat ttttgtgatg
     5041 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcagaaaggc ccacccgaag
     5101 gtgagccagg tgattacatt tgggccctca tcagaggttt tcaccgtcat caccgaaacg
     5161 cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc
     5221 ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat
     5281 aaagcgggcc atgttaaggg cggttttttc ctgtttggtc acttaccaat gcttaatcag
     5341 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5401 cgtgtagata actacgatgc gggagggctt accatctggc cccagtgctg caatgatacc
     5461 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccgggagggc
     5521 cgagcgcaga agtgatcctg caactttatc cgcctccatc cagtctatta attgttgccg
     5581 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     5641 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     5701 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     5761 tccgatcgtt gccagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     5821 gcataattct cttactgtca tgccatccgt gagatgcttt tctgtgactg gtgagtactc
     5881 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     5941 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6001 ttcggggcgt aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6061 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6121 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6181 cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga
     6241 tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaagtc aaaagcctcc
     6301 ggtcggaggc ttttgacttt ctgctatgga ggtcaggtat gattt
//
