LOCUS       sequence-298297-        6298 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 6298)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6298)
  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-m4046645
FEATURES             Location/Qualifiers
     source          1..6298
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     complement(2053..2071)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2422..3225
                     /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      3308..4290
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4368..4397
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4419..5006
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5018..5045)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5069..5260)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5261..6052,6053..6121))
                     /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     5884..5903
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6122..6224)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6225..6256)
                     /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 caagacttta aacaggcccc ttttcctttg tcgatatcat
     2041 gtaattagtt atgtcacgct tacattcacg ccctcctccc acatccgctc taaccgaaaa
     2101 ggaaggagtt agacaacctg aagtctaggt ccctatttat tttttttaat agttatgtta
     2161 gtattaagaa cgttatttat atttcaaatt tttctttttt ttctgtacaa agcggccgcc
     2221 gaaccactag tgcactgcag tacacggttt ccttgaaatt tttttgattc ggtaatctcc
     2281 gaacagaagg aagaacgaag gaaggagcac agacttagat tggtatatat acgcatatgt
     2341 agtgttgaag aaacatgaaa ttgcccagta ttcttaaccc aactgcacag aacaaaaacg
     2401 tgcaggaaac gaagataaat catgtcgaaa gctacatata aggaacgtgc tgctactcat
     2461 cctagtcctg ttgctgccaa gctatttaat atcatgcacg aaaagcaaac aaacttgtgt
     2521 gcttcattgg atgttcgtac caccaaggaa ttactggagt tagttgaagc attaggtccc
     2581 aaaatttgtt tactaaaaac acatgtggat atcttgactg atttttccat ggagggcaca
     2641 gttaagccgc taaaggcatt atccgccaag tacaattttt tactcttcga ggacagaaaa
     2701 tttgctgaca ttggtaatac agtcaaattg cagtactctg cgggtgtata cagaatagca
     2761 gaatgggcag acattacgaa tgcacacggt gtggtgggcc caggtattgt tagcggtttg
     2821 aagcaggcgg cagaagaagt aacaaaggaa cctagaggcc ttttgatgtt agcagaattg
     2881 tcatgcaagg gctccctatc tactggagaa tatactaagg gtactgttga cattgcgaag
     2941 agcgacaaag attttgttat cggctttatt gctcaaagag acatgggtgg aagagatgaa
     3001 ggttacgatt ggttgattat gacacccggt gtgggtttag atgacaaggg agatgcattg
     3061 ggtcaacagt atagaaccgt ggatgatgtg gtttctacag gatctgacat tattattgtt
     3121 ggaagaggac tatttgcaaa gggaagggat gctaaggtag agggtgaacg ttacagaaaa
     3181 gcaggctggg aagcatattt gagaagatgc ggccagcaaa actaaaaaac tgtattataa
     3241 gtaaatgcat gtatactaaa ctcacaaatt agagcttcaa tttaattata tcagttatta
     3301 cccgagtaac gaagcatctg tgcttcattt tgtagaacaa aaatgcaacg cgagagcgct
     3361 aatttttcaa acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgaaagc
     3421 gctattttac caacgaagaa tctgtgcttc atttttgtaa aacaaaaatg caacgcgaga
     3481 gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg
     3541 agagcgctat tttaccaaca aagaatctat acttcttttt tgttctacaa aaatgcatcc
     3601 cgagagcgct atttttctaa caaagcatct tagattactt tttttctcct ttgtgcgctc
     3661 tataatgcag tctcttgata actttttgca ctgtaggtcc gttaaggtta gaagaaggct
     3721 actttggtgt ctattttctc ttccataaaa aaagcctgac tccacttccc gcgtttactg
     3781 attactagcg aagctgcggg tgcatttttt caagataaag gcatccccga ttatattcta
     3841 taccgatgtg gattgcgcat actttgtgaa cagaaagtga tagcgttgat gattcttcat
     3901 tggtcagaaa attatgaacg gtttcttcta ttttgtctct atatactacg tataggaaat
     3961 gtttacattt tcgtattgtt ttcgattcac tctatgaata gttcttacta caattttttt
     4021 gtctaaagag taatactaga gataaacata aaaaatgtag aggtcgagtt tagatgcaag
     4081 ttcaaggagc gaaaggtgga tgggtaggtt atatagggat atagcacaga gatatatagc
     4141 aaagagatac ttttgagcaa tgtttgtgga agcggtattc gcaatatttt agtagctcgt
     4201 tacagtccgg tgcgtttttg gttttttgaa agtgcgtcat cagagcgctt ttggttttca
     4261 aaagcgctct gaagttccta tactttctag ctagagaata ggaacttccc gagcggccgc
     4321 gtgttacaag cttgactcca gcgtaactgg actgcaatca actcactggc tcaccttcac
     4381 gggtgggcct ttcttcggta gaaaatcaaa ggatcttctt gagatccttt ttttctgcgc
     4441 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     4501 caagagctac caactctttt tccgaggtaa ctggcttcag cagagcgcag ataccaaata
     4561 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     4621 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     4681 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     4741 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     4801 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     4861 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggta aacgcctggt
     4921 atctttatag tcctgtcggg tttcgccacc tctgacttga gcatcgattt ttgtgatgct
     4981 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc agaaaggccc acccgaaggt
     5041 gagccaggtg attacatttg ggccctcatc agaggttttc accgtcatca ccgaaacgcg
     5101 cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct
     5161 gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa
     5221 agcgggccat gttaagggcg gttttttcct gtttggtcac ttaccaatgc ttaatcagtg
     5281 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     5341 tgtagataac tacgatgcgg gagggcttac catctggccc cagtgctgca atgataccgc
     5401 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc gggagggccg
     5461 agcgcagaag tgatcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     5521 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     5581 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     5641 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     5701 cgatcgttgc cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     5761 ataattctct tactgtcatg ccatccgtga gatgcttttc tgtgactggt gagtactcaa
     5821 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     5881 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     5941 cggggcgtaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6001 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     6061 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6121 ttttagcttc cttagctcct gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc
     6181 ttatttcatt atggtgaaag ttggaacctc ttacgtgccg atcaagtcaa aagcctccgg
     6241 tcggaggctt ttgactttct gctatggagg tcaggtatga tttaaatggt cagttaga
//
