LOCUS       sequence-298270-        6304 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 6304)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6304)
  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-m4046666
FEATURES             Location/Qualifiers
     source          1..6304
                     /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(2059..2077)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2428..3231
                     /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      3314..4296
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4374..4403
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4425..5012
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5024..5051)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5075..5266)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5267..6058,6059..6127))
                     /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     5890..5909
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6128..6230)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6231..6262)
                     /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 atggccgaga atgtcacagg cctattcaag gattgctcga aagtgatcac
      481 cggtttacac cctacgcagg ctccaactca tttgtccgtt gacaccaagt ttaagactga
      541 aggattgtgt gttgacatac caggtattcc caaagatatg acttaccgta ggctgatttc
      601 tatgatgggt ttcaagatga actatcaagt taacggttac ccaaatatgt ttatcaccag
      661 agaagaagct attagacacg tacgtgcatg gatcggtttc gatgtcgaag gttgtcatgc
      721 tactagagaa gcggtcggta caaacttgcc attacaactt ggcttttcaa ccggtgttaa
      781 tttggttgct gttcctacag ggtatgtcga cactccaaac aacacggatt tctctcgagt
      841 gtccgccaag ccaccacctg gtgatcaatt caaacacttg atcccgctca tgtacaaggg
      901 tttgccttgg aacgttgtca gaattaaaat agtacaaatg ttatctgata ctttgaagaa
      961 tctgagtgac agagttgtct tcgtcctttg ggctcacggt tttgagctaa ccagcatgaa
     1021 gtactttgtg aagattggac cagaacggac ttgttgcttg tgtgacagaa gagccacttg
     1081 tttctccacc gctagcgata catatgcatg ttggcatcat tctattggtt tcgactacgt
     1141 ttacaacccc tttatgatcg acgttcaaca gtggggtttc accggtaatt tgcaatcgaa
     1201 ccacgacttg tattgtcaag ttcacggcaa tgctcacgtg gcttcatgcg atgctattat
     1261 gacaaggtgt ttagccgtcc atgaatgttt cgttaaaaga gtagactgga ctatcgaata
     1321 cccaattatc ggtgatgaac tcaagattaa cgctgcctgt agaaaggtcc aacatatggt
     1381 tgtaaaggca gctctattag ctgacaaatt tccagttttg cacgatatag ggaatccaaa
     1441 ggccataaag tgtgttccac aagctgacgt cgaatggaaa ttctacgatg cgcaaccctg
     1501 ctctgataaa gcttataaga tcgaggaatt gttttactct tacgctaccc actccgataa
     1561 gttcactgac ggtgtgtgtc ttttctggaa ctgcaatgtt gatagatatc cagcaaactc
     1621 catcgtttgt agattcgaca cgcgagtctt gagcaacttg aacttaccag gttgtgatgg
     1681 tggttccttg tacgtcaata aacatgcatt tcacactccc gctttcgaca agtctgcttt
     1741 tgttaattta aagcaattgc cttttttcta ttactctgat agtccatgcg aatctcatgg
     1801 taagcaagtc gtttccgaca ttgattacgt accactgaag tcagccacct gtataacaag
     1861 atgtaacctt ggaggtgctg tctgccgcca ccatgctaac gagtacagac tatacttgga
     1921 tgcctataac atgatgattt cagccggttt ctctttatgg gtttacaagc agtttgacac
     1981 ttacaacttg tggaacacct tcacgcgttt gcaataaaca ggcccctttt cctttgtcga
     2041 tatcatgtaa ttagttatgt cacgcttaca ttcacgccct cctcccacat ccgctctaac
     2101 cgaaaaggaa ggagttagac aacctgaagt ctaggtccct atttattttt tttaatagtt
     2161 atgttagtat taagaacgtt atttatattt caaatttttc ttttttttct gtacaaagcg
     2221 gccgccgaac cactagtgca ctgcagtaca cggtttcctt gaaatttttt tgattcggta
     2281 atctccgaac agaaggaaga acgaaggaag gagcacagac ttagattggt atatatacgc
     2341 atatgtagtg ttgaagaaac atgaaattgc ccagtattct taacccaact gcacagaaca
     2401 aaaacgtgca ggaaacgaag ataaatcatg tcgaaagcta catataagga acgtgctgct
     2461 actcatccta gtcctgttgc tgccaagcta tttaatatca tgcacgaaaa gcaaacaaac
     2521 ttgtgtgctt cattggatgt tcgtaccacc aaggaattac tggagttagt tgaagcatta
     2581 ggtcccaaaa tttgtttact aaaaacacat gtggatatct tgactgattt ttccatggag
     2641 ggcacagtta agccgctaaa ggcattatcc gccaagtaca attttttact cttcgaggac
     2701 agaaaatttg ctgacattgg taatacagtc aaattgcagt actctgcggg tgtatacaga
     2761 atagcagaat gggcagacat tacgaatgca cacggtgtgg tgggcccagg tattgttagc
     2821 ggtttgaagc aggcggcaga agaagtaaca aaggaaccta gaggcctttt gatgttagca
     2881 gaattgtcat gcaagggctc cctatctact ggagaatata ctaagggtac tgttgacatt
     2941 gcgaagagcg acaaagattt tgttatcggc tttattgctc aaagagacat gggtggaaga
     3001 gatgaaggtt acgattggtt gattatgaca cccggtgtgg gtttagatga caagggagat
     3061 gcattgggtc aacagtatag aaccgtggat gatgtggttt ctacaggatc tgacattatt
     3121 attgttggaa gaggactatt tgcaaaggga agggatgcta aggtagaggg tgaacgttac
     3181 agaaaagcag gctgggaagc atatttgaga agatgcggcc agcaaaacta aaaaactgta
     3241 ttataagtaa atgcatgtat actaaactca caaattagag cttcaattta attatatcag
     3301 ttattacccg agtaacgaag catctgtgct tcattttgta gaacaaaaat gcaacgcgag
     3361 agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc
     3421 gaaagcgcta ttttaccaac gaagaatctg tgcttcattt ttgtaaaaca aaaatgcaac
     3481 gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc
     3541 aacgcgagag cgctatttta ccaacaaaga atctatactt cttttttgtt ctacaaaaat
     3601 gcatcccgag agcgctattt ttctaacaaa gcatcttaga ttactttttt tctcctttgt
     3661 gcgctctata atgcagtctc ttgataactt tttgcactgt aggtccgtta aggttagaag
     3721 aaggctactt tggtgtctat tttctcttcc ataaaaaaag cctgactcca cttcccgcgt
     3781 ttactgatta ctagcgaagc tgcgggtgca ttttttcaag ataaaggcat ccccgattat
     3841 attctatacc gatgtggatt gcgcatactt tgtgaacaga aagtgatagc gttgatgatt
     3901 cttcattggt cagaaaatta tgaacggttt cttctatttt gtctctatat actacgtata
     3961 ggaaatgttt acattttcgt attgttttcg attcactcta tgaatagttc ttactacaat
     4021 ttttttgtct aaagagtaat actagagata aacataaaaa atgtagaggt cgagtttaga
     4081 tgcaagttca aggagcgaaa ggtggatggg taggttatat agggatatag cacagagata
     4141 tatagcaaag agatactttt gagcaatgtt tgtggaagcg gtattcgcaa tattttagta
     4201 gctcgttaca gtccggtgcg tttttggttt tttgaaagtg cgtcatcaga gcgcttttgg
     4261 ttttcaaaag cgctctgaag ttcctatact ttctagctag agaataggaa cttcccgagc
     4321 ggccgcgtgt tacaagcttg actccagcgt aactggactg caatcaactc actggctcac
     4381 cttcacgggt gggcctttct tcggtagaaa atcaaaggat cttcttgaga tccttttttt
     4441 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     4501 ccggatcaag agctaccaac tctttttccg aggtaactgg cttcagcaga gcgcagatac
     4561 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     4621 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     4681 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     4741 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     4801 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     4861 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca ggggtaaacg
     4921 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcat cgatttttgt
     4981 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcagaa aggcccaccc
     5041 gaaggtgagc caggtgatta catttgggcc ctcatcagag gttttcaccg tcatcaccga
     5101 aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt
     5161 ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc
     5221 tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcacttac caatgcttaa
     5281 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5341 ccgtcgtgta gataactacg atgcgggagg gcttaccatc tggccccagt gctgcaatga
     5401 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggga
     5461 gggccgagcg cagaagtgat cctgcaactt tatccgcctc catccagtct attaattgtt
     5521 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5581 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5641 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5701 gtcctccgat cgttgccaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5761 cactgcataa ttctcttact gtcatgccat ccgtgagatg cttttctgtg actggtgagt
     5821 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5881 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5941 gttcttcggg gcgtaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6001 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6061 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6121 tactcatttt agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta
     6181 gtgatcttat ttcattatgg tgaaagttgg aacctcttac gtgccgatca agtcaaaagc
     6241 ctccggtcgg aggcttttga ctttctgcta tggaggtcag gtatgattta aatggtcagt
     6301 taga
//
