LOCUS       sequence-297626-        6433 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast expression plasmid for SARS-CoV-2 nsp8.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6433)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6433)
  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-m4046256
FEATURES             Location/Qualifiers
     source          1..6433
                     /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             1028..1045
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2188..2206)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2557..3360
                     /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      3443..4425
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4503..4532
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4554..5141
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5153..5180)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5204..5395)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5396..6187,6188..6256))
                     /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     6019..6038
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6257..6359)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6360..6391)
                     /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 atggctatcg ccagtgaatt ttcctcatta cctagctatg cagctttcgc
      481 tacggctcag gaggcctacg aacaagctgt agccaacggc gactctgaag ttgttcttaa
      541 gaaattgaag aagtctttga atgtcgcgaa atctgaattt gatagagatg ctgctatgca
      601 acgtaagcta gaaaagatgg ctgaccaagc aatgacccaa atgtacaaac aagccagaag
      661 tgaggacaag cgagctaagg tgacttccgc tatgcaaaca atgctgttca ctatgttgag
      721 aaagttggat aacgatgctc tgaacaatat tataaacaat gccagagatg gttgtgtccc
      781 cctcaacatt atcccattaa ccactgccgc taaattgatg gttgttattc cggattataa
      841 tacatacaaa aacacttgcg acggtaccac cttcacatac gcctctgctc tttgggaaat
      901 ccaacaggta gtcgacgctg actccaagat agttcaattg agcgaaattt ctatggataa
      961 ctcgccaaac ctagcttggc cattaatcgt cactgcgttg agagctaata gtgcggttaa
     1021 gttgcaactc gtgccgagag gttctaaaat cgaggaaggt aagctagtca tttggataaa
     1081 tggagataag ggttacaacg gtttggcaga agtaggcaag aaatttgaaa aggacacagg
     1141 tatcaaggtg acggttgaac atccagacaa gttagaagaa aaattccctc aagttgctgc
     1201 tactggtgat ggtcccgaca ttattttctg ggcccacgat cgttttggtg gatatgctca
     1261 gtcaggtttg ctggctgaga tcaccccaga caaggccttc caagataaat tgtacccatt
     1321 cacttgggat gctgtcagat acaacggtaa gcttatagca tatccaattg ctgttgaagc
     1381 cttatcgttg atctacaata aggacttgtt acctaaccca ccaaagacat gggaagaaat
     1441 tccagctttg gataaagagc taaaggccaa gggaaaaagc gctttgatgt ttaacttaca
     1501 agaaccttat ttcacctggc cattaatcgc tgctgacggt ggttacgcat ttaaatacga
     1561 aaacggtaag tatgacatta aggatgtcgg cgttgataat gctggtgcaa aggctggttt
     1621 gaccttcttg gttgatttaa taaagaataa acacatgaac gctgacactg actactctat
     1681 cgccgaagcc gctttcaaca agggtgaaac tgctatgacc ataaatggtc catgggcctg
     1741 gagtaacatt gatacttcta aagtaaacta tggtgtcact gttttgccta cattcaaagg
     1801 ccaaccatcc aagccctttg tcggtgttct atccgctggc atcaatgctg cttctccaaa
     1861 caaggagcta gctaaagaat ttttggaaaa ttacttgtta actgacgaag gtttggaggc
     1921 cgttaacaag gacaagccat tgggtgctgt cgctttgaaa tcttacgaag aagaattggc
     1981 taaggatccc agaattgccg ctaccatgga gaacgcccaa aagggtgaaa ttatgccaaa
     2041 catcccacaa atgtcagcct tctggtacgc agttaggact gccgtgatta atgctgcttc
     2101 cggtagacaa accgtcgacg aagcattaaa agatgctcaa acgtaaacag gccccttttc
     2161 ctttgtcgat atcatgtaat tagttatgtc acgcttacat tcacgccctc ctcccacatc
     2221 cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt
     2281 ttaatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg
     2341 tacaaagcgg ccgccgaacc actagtgcac tgcagtacac ggtttccttg aaattttttt
     2401 gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg agcacagact tagattggta
     2461 tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt aacccaactg
     2521 cacagaacaa aaacgtgcag gaaacgaaga taaatcatgt cgaaagctac atataaggaa
     2581 cgtgctgcta ctcatcctag tcctgttgct gccaagctat ttaatatcat gcacgaaaag
     2641 caaacaaact tgtgtgcttc attggatgtt cgtaccacca aggaattact ggagttagtt
     2701 gaagcattag gtcccaaaat ttgtttacta aaaacacatg tggatatctt gactgatttt
     2761 tccatggagg gcacagttaa gccgctaaag gcattatccg ccaagtacaa ttttttactc
     2821 ttcgaggaca gaaaatttgc tgacattggt aatacagtca aattgcagta ctctgcgggt
     2881 gtatacagaa tagcagaatg ggcagacatt acgaatgcac acggtgtggt gggcccaggt
     2941 attgttagcg gtttgaagca ggcggcagaa gaagtaacaa aggaacctag aggccttttg
     3001 atgttagcag aattgtcatg caagggctcc ctatctactg gagaatatac taagggtact
     3061 gttgacattg cgaagagcga caaagatttt gttatcggct ttattgctca aagagacatg
     3121 ggtggaagag atgaaggtta cgattggttg attatgacac ccggtgtggg tttagatgac
     3181 aagggagatg cattgggtca acagtataga accgtggatg atgtggtttc tacaggatct
     3241 gacattatta ttgttggaag aggactattt gcaaagggaa gggatgctaa ggtagagggt
     3301 gaacgttaca gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa
     3361 aaaactgtat tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa
     3421 ttatatcagt tattacccga gtaacgaagc atctgtgctt cattttgtag aacaaaaatg
     3481 caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa
     3541 atgcaacgcg aaagcgctat tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa
     3601 aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa
     3661 cagaaatgca acgcgagagc gctattttac caacaaagaa tctatacttc ttttttgttc
     3721 tacaaaaatg catcccgaga gcgctatttt tctaacaaag catcttagat tacttttttt
     3781 ctcctttgtg cgctctataa tgcagtctct tgataacttt ttgcactgta ggtccgttaa
     3841 ggttagaaga aggctacttt ggtgtctatt ttctcttcca taaaaaaagc ctgactccac
     3901 ttcccgcgtt tactgattac tagcgaagct gcgggtgcat tttttcaaga taaaggcatc
     3961 cccgattata ttctataccg atgtggattg cgcatacttt gtgaacagaa agtgatagcg
     4021 ttgatgattc ttcattggtc agaaaattat gaacggtttc ttctattttg tctctatata
     4081 ctacgtatag gaaatgttta cattttcgta ttgttttcga ttcactctat gaatagttct
     4141 tactacaatt tttttgtcta aagagtaata ctagagataa acataaaaaa tgtagaggtc
     4201 gagtttagat gcaagttcaa ggagcgaaag gtggatgggt aggttatata gggatatagc
     4261 acagagatat atagcaaaga gatacttttg agcaatgttt gtggaagcgg tattcgcaat
     4321 attttagtag ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc gtcatcagag
     4381 cgcttttggt tttcaaaagc gctctgaagt tcctatactt tctagctaga gaataggaac
     4441 ttcccgagcg gccgcgtgtt acaagcttga ctccagcgta actggactgc aatcaactca
     4501 ctggctcacc ttcacgggtg ggcctttctt cggtagaaaa tcaaaggatc ttcttgagat
     4561 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     4621 gtttgtttgc cggatcaaga gctaccaact ctttttccga ggtaactggc ttcagcagag
     4681 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     4741 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     4801 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     4861 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     4921 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     4981 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     5041 gggtaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcatc
     5101 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcagaaa
     5161 ggcccacccg aaggtgagcc aggtgattac atttgggccc tcatcagagg ttttcaccgt
     5221 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt
     5281 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg
     5341 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcacttacc
     5401 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     5461 cctgactccc cgtcgtgtag ataactacga tgcgggaggg cttaccatct ggccccagtg
     5521 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     5581 cagccgggag ggccgagcgc agaagtgatc ctgcaacttt atccgcctcc atccagtcta
     5641 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     5701 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     5761 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     5821 gctccttcgg tcctccgatc gttgccagaa gtaagttggc cgcagtgtta tcactcatgg
     5881 ttatggcagc actgcataat tctcttactg tcatgccatc cgtgagatgc ttttctgtga
     5941 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     6001 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6061 ttggaaaacg ttcttcgggg cgtaaactct caaggatctt accgctgttg agatccagtt
     6121 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     6181 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     6241 aatgttgaat actcatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata
     6301 cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa
     6361 gtcaaaagcc tccggtcgga ggcttttgac tttctgctat ggaggtcagg tatgatttaa
     6421 atggtcagtt aga
//
