LOCUS       sequence-291560-        6346 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 6346)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6346)
  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-m4046224
FEATURES             Location/Qualifiers
     source          1..6346
                     /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"
     CDS             2033..2056
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(2101..2119)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2470..3273
                     /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      3356..4338
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4416..4445
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4467..5054
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5066..5093)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5117..5308)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5309..6100,6101..6169))
                     /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     5932..5951
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6170..6272)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6273..6304)
                     /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 cttggagcca
     2041 tccgcagttt gaaaaataaa caggcccctt ttcctttgtc gatatcatgt aattagttat
     2101 gtcacgctta cattcacgcc ctcctcccac atccgctcta accgaaaagg aaggagttag
     2161 acaacctgaa gtctaggtcc ctatttattt tttttaatag ttatgttagt attaagaacg
     2221 ttatttatat ttcaaatttt tctttttttt ctgtacaaag cggccgccga accactagtg
     2281 cactgcagta cacggtttcc ttgaaatttt tttgattcgg taatctccga acagaaggaa
     2341 gaacgaagga aggagcacag acttagattg gtatatatac gcatatgtag tgttgaagaa
     2401 acatgaaatt gcccagtatt cttaacccaa ctgcacagaa caaaaacgtg caggaaacga
     2461 agataaatca tgtcgaaagc tacatataag gaacgtgctg ctactcatcc tagtcctgtt
     2521 gctgccaagc tatttaatat catgcacgaa aagcaaacaa acttgtgtgc ttcattggat
     2581 gttcgtacca ccaaggaatt actggagtta gttgaagcat taggtcccaa aatttgttta
     2641 ctaaaaacac atgtggatat cttgactgat ttttccatgg agggcacagt taagccgcta
     2701 aaggcattat ccgccaagta caatttttta ctcttcgagg acagaaaatt tgctgacatt
     2761 ggtaatacag tcaaattgca gtactctgcg ggtgtataca gaatagcaga atgggcagac
     2821 attacgaatg cacacggtgt ggtgggccca ggtattgtta gcggtttgaa gcaggcggca
     2881 gaagaagtaa caaaggaacc tagaggcctt ttgatgttag cagaattgtc atgcaagggc
     2941 tccctatcta ctggagaata tactaagggt actgttgaca ttgcgaagag cgacaaagat
     3001 tttgttatcg gctttattgc tcaaagagac atgggtggaa gagatgaagg ttacgattgg
     3061 ttgattatga cacccggtgt gggtttagat gacaagggag atgcattggg tcaacagtat
     3121 agaaccgtgg atgatgtggt ttctacagga tctgacatta ttattgttgg aagaggacta
     3181 tttgcaaagg gaagggatgc taaggtagag ggtgaacgtt acagaaaagc aggctgggaa
     3241 gcatatttga gaagatgcgg ccagcaaaac taaaaaactg tattataagt aaatgcatgt
     3301 atactaaact cacaaattag agcttcaatt taattatatc agttattacc cgagtaacga
     3361 agcatctgtg cttcattttg tagaacaaaa atgcaacgcg agagcgctaa tttttcaaac
     3421 aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgaaagcgc tattttacca
     3481 acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca acgcgagagc gctaattttt
     3541 caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgag agcgctattt
     3601 taccaacaaa gaatctatac ttcttttttg ttctacaaaa atgcatcccg agagcgctat
     3661 ttttctaaca aagcatctta gattactttt tttctccttt gtgcgctcta taatgcagtc
     3721 tcttgataac tttttgcact gtaggtccgt taaggttaga agaaggctac tttggtgtct
     3781 attttctctt ccataaaaaa agcctgactc cacttcccgc gtttactgat tactagcgaa
     3841 gctgcgggtg cattttttca agataaaggc atccccgatt atattctata ccgatgtgga
     3901 ttgcgcatac tttgtgaaca gaaagtgata gcgttgatga ttcttcattg gtcagaaaat
     3961 tatgaacggt ttcttctatt ttgtctctat atactacgta taggaaatgt ttacattttc
     4021 gtattgtttt cgattcactc tatgaatagt tcttactaca atttttttgt ctaaagagta
     4081 atactagaga taaacataaa aaatgtagag gtcgagttta gatgcaagtt caaggagcga
     4141 aaggtggatg ggtaggttat atagggatat agcacagaga tatatagcaa agagatactt
     4201 ttgagcaatg tttgtggaag cggtattcgc aatattttag tagctcgtta cagtccggtg
     4261 cgtttttggt tttttgaaag tgcgtcatca gagcgctttt ggttttcaaa agcgctctga
     4321 agttcctata ctttctagct agagaatagg aacttcccga gcggccgcgt gttacaagct
     4381 tgactccagc gtaactggac tgcaatcaac tcactggctc accttcacgg gtgggccttt
     4441 cttcggtaga aaatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     4501 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     4561 actctttttc cgaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     4621 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4681 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4741 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4801 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4861 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4921 tcggaacagg agagcgcacg agggagcttc caggggtaaa cgcctggtat ctttatagtc
     4981 ctgtcgggtt tcgccacctc tgacttgagc atcgattttt gtgatgctcg tcaggggggc
     5041 ggagcctatg gaaaaacgcc agcaacgcag aaaggcccac ccgaaggtga gccaggtgat
     5101 tacatttggg ccctcatcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc
     5161 ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt
     5221 ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt
     5281 taagggcggt tttttcctgt ttggtcactt accaatgctt aatcagtgag gcacctatct
     5341 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     5401 cgatgcggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     5461 caccggctcc agatttatca gcaataaacc agccagccgg gagggccgag cgcagaagtg
     5521 atcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     5581 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     5641 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     5701 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgcca
     5761 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     5821 ctgtcatgcc atccgtgaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     5881 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     5941 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgtaaac
     6001 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     6061 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     6121 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcatt ttagcttcct
     6181 tagctcctga aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat
     6241 ggtgaaagtt ggaacctctt acgtgccgat caagtcaaaa gcctccggtc ggaggctttt
     6301 gactttctgc tatggaggtc aggtatgatt taaatggtca gttaga
//
