LOCUS       sequence-297988-        7420 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 7420)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 7420)
  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-m4046271
FEATURES             Location/Qualifiers
     source          1..7420
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             2015..2032
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(3175..3193)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             3544..4347
                     /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      4430..5412
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      5490..5519
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      5541..6128
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(6140..6167)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(6191..6382)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6383..7174,7175..7243))
                     /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     7006..7025
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7244..7346)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(7347..7378)
                     /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 gcaactcgtg ccgagaggtt ctaaaatcga
     2041 ggaaggtaag ctagtcattt ggataaatgg agataagggt tacaacggtt tggcagaagt
     2101 aggcaagaaa tttgaaaagg acacaggtat caaggtgacg gttgaacatc cagacaagtt
     2161 agaagaaaaa ttccctcaag ttgctgctac tggtgatggt cccgacatta ttttctgggc
     2221 ccacgatcgt tttggtggat atgctcagtc aggtttgctg gctgagatca ccccagacaa
     2281 ggccttccaa gataaattgt acccattcac ttgggatgct gtcagataca acggtaagct
     2341 tatagcatat ccaattgctg ttgaagcctt atcgttgatc tacaataagg acttgttacc
     2401 taacccacca aagacatggg aagaaattcc agctttggat aaagagctaa aggccaaggg
     2461 aaaaagcgct ttgatgttta acttacaaga accttatttc acctggccat taatcgctgc
     2521 tgacggtggt tacgcattta aatacgaaaa cggtaagtat gacattaagg atgtcggcgt
     2581 tgataatgct ggtgcaaagg ctggtttgac cttcttggtt gatttaataa agaataaaca
     2641 catgaacgct gacactgact actctatcgc cgaagccgct ttcaacaagg gtgaaactgc
     2701 tatgaccata aatggtccat gggcctggag taacattgat acttctaaag taaactatgg
     2761 tgtcactgtt ttgcctacat tcaaaggcca accatccaag ccctttgtcg gtgttctatc
     2821 cgctggcatc aatgctgctt ctccaaacaa ggagctagct aaagaatttt tggaaaatta
     2881 cttgttaact gacgaaggtt tggaggccgt taacaaggac aagccattgg gtgctgtcgc
     2941 tttgaaatct tacgaagaag aattggctaa ggatcccaga attgccgcta ccatggagaa
     3001 cgcccaaaag ggtgaaatta tgccaaacat cccacaaatg tcagccttct ggtacgcagt
     3061 taggactgcc gtgattaatg ctgcttccgg tagacaaacc gtcgacgaag cattaaaaga
     3121 tgctcaaacg taaacaggcc ccttttcctt tgtcgatatc atgtaattag ttatgtcacg
     3181 cttacattca cgccctcctc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc
     3241 tgaagtctag gtccctattt atttttttta atagttatgt tagtattaag aacgttattt
     3301 atatttcaaa tttttctttt ttttctgtac aaagcggccg ccgaaccact agtgcactgc
     3361 agtacacggt ttccttgaaa tttttttgat tcggtaatct ccgaacagaa ggaagaacga
     3421 aggaaggagc acagacttag attggtatat atacgcatat gtagtgttga agaaacatga
     3481 aattgcccag tattcttaac ccaactgcac agaacaaaaa cgtgcaggaa acgaagataa
     3541 atcatgtcga aagctacata taaggaacgt gctgctactc atcctagtcc tgttgctgcc
     3601 aagctattta atatcatgca cgaaaagcaa acaaacttgt gtgcttcatt ggatgttcgt
     3661 accaccaagg aattactgga gttagttgaa gcattaggtc ccaaaatttg tttactaaaa
     3721 acacatgtgg atatcttgac tgatttttcc atggagggca cagttaagcc gctaaaggca
     3781 ttatccgcca agtacaattt tttactcttc gaggacagaa aatttgctga cattggtaat
     3841 acagtcaaat tgcagtactc tgcgggtgta tacagaatag cagaatgggc agacattacg
     3901 aatgcacacg gtgtggtggg cccaggtatt gttagcggtt tgaagcaggc ggcagaagaa
     3961 gtaacaaagg aacctagagg ccttttgatg ttagcagaat tgtcatgcaa gggctcccta
     4021 tctactggag aatatactaa gggtactgtt gacattgcga agagcgacaa agattttgtt
     4081 atcggcttta ttgctcaaag agacatgggt ggaagagatg aaggttacga ttggttgatt
     4141 atgacacccg gtgtgggttt agatgacaag ggagatgcat tgggtcaaca gtatagaacc
     4201 gtggatgatg tggtttctac aggatctgac attattattg ttggaagagg actatttgca
     4261 aagggaaggg atgctaaggt agagggtgaa cgttacagaa aagcaggctg ggaagcatat
     4321 ttgagaagat gcggccagca aaactaaaaa actgtattat aagtaaatgc atgtatacta
     4381 aactcacaaa ttagagcttc aatttaatta tatcagttat tacccgagta acgaagcatc
     4441 tgtgcttcat tttgtagaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa
     4501 tctgagctgc atttttacag aacagaaatg caacgcgaaa gcgctatttt accaacgaag
     4561 aatctgtgct tcatttttgt aaaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca
     4621 aagaatctga gctgcatttt tacagaacag aaatgcaacg cgagagcgct attttaccaa
     4681 caaagaatct atacttcttt tttgttctac aaaaatgcat cccgagagcg ctatttttct
     4741 aacaaagcat cttagattac tttttttctc ctttgtgcgc tctataatgc agtctcttga
     4801 taactttttg cactgtaggt ccgttaaggt tagaagaagg ctactttggt gtctattttc
     4861 tcttccataa aaaaagcctg actccacttc ccgcgtttac tgattactag cgaagctgcg
     4921 ggtgcatttt ttcaagataa aggcatcccc gattatattc tataccgatg tggattgcgc
     4981 atactttgtg aacagaaagt gatagcgttg atgattcttc attggtcaga aaattatgaa
     5041 cggtttcttc tattttgtct ctatatacta cgtataggaa atgtttacat tttcgtattg
     5101 ttttcgattc actctatgaa tagttcttac tacaattttt ttgtctaaag agtaatacta
     5161 gagataaaca taaaaaatgt agaggtcgag tttagatgca agttcaagga gcgaaaggtg
     5221 gatgggtagg ttatataggg atatagcaca gagatatata gcaaagagat acttttgagc
     5281 aatgtttgtg gaagcggtat tcgcaatatt ttagtagctc gttacagtcc ggtgcgtttt
     5341 tggttttttg aaagtgcgtc atcagagcgc ttttggtttt caaaagcgct ctgaagttcc
     5401 tatactttct agctagagaa taggaacttc ccgagcggcc gcgtgttaca agcttgactc
     5461 cagcgtaact ggactgcaat caactcactg gctcaccttc acgggtgggc ctttcttcgg
     5521 tagaaaatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5581 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5641 tttccgaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     5701 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     5761 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     5821 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     5881 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     5941 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     6001 caggagagcg cacgagggag cttccagggg taaacgcctg gtatctttat agtcctgtcg
     6061 ggtttcgcca cctctgactt gagcatcgat ttttgtgatg ctcgtcaggg gggcggagcc
     6121 tatggaaaaa cgccagcaac gcagaaaggc ccacccgaag gtgagccagg tgattacatt
     6181 tgggccctca tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa
     6241 gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct
     6301 cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg
     6361 cggttttttc ctgtttggtc acttaccaat gcttaatcag tgaggcacct atctcagcga
     6421 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatgc
     6481 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     6541 ctccagattt atcagcaata aaccagccag ccgggagggc cgagcgcaga agtgatcctg
     6601 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     6661 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     6721 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     6781 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gccagaagta
     6841 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     6901 tgccatccgt gagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     6961 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     7021 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcgt aaactctcaa
     7081 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     7141 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     7201 caaaaaaggg aataagggcg acacggaaat gttgaatact cattttagct tccttagctc
     7261 ctgaaaatct cgataactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa
     7321 agttggaacc tcttacgtgc cgatcaagtc aaaagcctcc ggtcggaggc ttttgacttt
     7381 ctgctatgga ggtcaggtat gatttaaatg gtcagttaga
//
