LOCUS       sequence-291557-        5353 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 5353)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5353)
  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-m4046477
FEATURES             Location/Qualifiers
     source          1..5353
                     /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"
     CDS             1046..1063
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1108..1126)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1477..2280
                     /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      2363..3345
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3423..3452
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3474..4061
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4073..4100)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4124..4315)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4316..5107,5108..5176))
                     /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     4939..4958
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5177..5279)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5280..5311)
                     /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 gttctcacca tcatcaccat cattaaacag gccccttttc
     1081 ctttgtcgat atcatgtaat tagttatgtc acgcttacat tcacgccctc ctcccacatc
     1141 cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt
     1201 ttaatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg
     1261 tacaaagcgg ccgccgaacc actagtgcac tgcagtacac ggtttccttg aaattttttt
     1321 gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg agcacagact tagattggta
     1381 tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt aacccaactg
     1441 cacagaacaa aaacgtgcag gaaacgaaga taaatcatgt cgaaagctac atataaggaa
     1501 cgtgctgcta ctcatcctag tcctgttgct gccaagctat ttaatatcat gcacgaaaag
     1561 caaacaaact tgtgtgcttc attggatgtt cgtaccacca aggaattact ggagttagtt
     1621 gaagcattag gtcccaaaat ttgtttacta aaaacacatg tggatatctt gactgatttt
     1681 tccatggagg gcacagttaa gccgctaaag gcattatccg ccaagtacaa ttttttactc
     1741 ttcgaggaca gaaaatttgc tgacattggt aatacagtca aattgcagta ctctgcgggt
     1801 gtatacagaa tagcagaatg ggcagacatt acgaatgcac acggtgtggt gggcccaggt
     1861 attgttagcg gtttgaagca ggcggcagaa gaagtaacaa aggaacctag aggccttttg
     1921 atgttagcag aattgtcatg caagggctcc ctatctactg gagaatatac taagggtact
     1981 gttgacattg cgaagagcga caaagatttt gttatcggct ttattgctca aagagacatg
     2041 ggtggaagag atgaaggtta cgattggttg attatgacac ccggtgtggg tttagatgac
     2101 aagggagatg cattgggtca acagtataga accgtggatg atgtggtttc tacaggatct
     2161 gacattatta ttgttggaag aggactattt gcaaagggaa gggatgctaa ggtagagggt
     2221 gaacgttaca gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa
     2281 aaaactgtat tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa
     2341 ttatatcagt tattacccga gtaacgaagc atctgtgctt cattttgtag aacaaaaatg
     2401 caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa
     2461 atgcaacgcg aaagcgctat tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa
     2521 aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa
     2581 cagaaatgca acgcgagagc gctattttac caacaaagaa tctatacttc ttttttgttc
     2641 tacaaaaatg catcccgaga gcgctatttt tctaacaaag catcttagat tacttttttt
     2701 ctcctttgtg cgctctataa tgcagtctct tgataacttt ttgcactgta ggtccgttaa
     2761 ggttagaaga aggctacttt ggtgtctatt ttctcttcca taaaaaaagc ctgactccac
     2821 ttcccgcgtt tactgattac tagcgaagct gcgggtgcat tttttcaaga taaaggcatc
     2881 cccgattata ttctataccg atgtggattg cgcatacttt gtgaacagaa agtgatagcg
     2941 ttgatgattc ttcattggtc agaaaattat gaacggtttc ttctattttg tctctatata
     3001 ctacgtatag gaaatgttta cattttcgta ttgttttcga ttcactctat gaatagttct
     3061 tactacaatt tttttgtcta aagagtaata ctagagataa acataaaaaa tgtagaggtc
     3121 gagtttagat gcaagttcaa ggagcgaaag gtggatgggt aggttatata gggatatagc
     3181 acagagatat atagcaaaga gatacttttg agcaatgttt gtggaagcgg tattcgcaat
     3241 attttagtag ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc gtcatcagag
     3301 cgcttttggt tttcaaaagc gctctgaagt tcctatactt tctagctaga gaataggaac
     3361 ttcccgagcg gccgcgtgtt acaagcttga ctccagcgta actggactgc aatcaactca
     3421 ctggctcacc ttcacgggtg ggcctttctt cggtagaaaa tcaaaggatc ttcttgagat
     3481 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     3541 gtttgtttgc cggatcaaga gctaccaact ctttttccga ggtaactggc ttcagcagag
     3601 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     3661 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     3721 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     3781 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     3841 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     3901 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     3961 gggtaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcatc
     4021 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcagaaa
     4081 ggcccacccg aaggtgagcc aggtgattac atttgggccc tcatcagagg ttttcaccgt
     4141 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt
     4201 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg
     4261 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcacttacc
     4321 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     4381 cctgactccc cgtcgtgtag ataactacga tgcgggaggg cttaccatct ggccccagtg
     4441 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     4501 cagccgggag ggccgagcgc agaagtgatc ctgcaacttt atccgcctcc atccagtcta
     4561 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     4621 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     4681 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     4741 gctccttcgg tcctccgatc gttgccagaa gtaagttggc cgcagtgtta tcactcatgg
     4801 ttatggcagc actgcataat tctcttactg tcatgccatc cgtgagatgc ttttctgtga
     4861 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     4921 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     4981 ttggaaaacg ttcttcgggg cgtaaactct caaggatctt accgctgttg agatccagtt
     5041 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     5101 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     5161 aatgttgaat actcatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata
     5221 cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa
     5281 gtcaaaagcc tccggtcgga ggcttttgac tttctgctat ggaggtcagg tatgatttaa
     5341 atggtcagtt aga
//
