LOCUS       sequence-292509-        5400 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 5400)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5400)
  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-m4046185
FEATURES             Location/Qualifiers
     source          1..5400
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        58..499
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    58..175
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     470..493
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             1102..1119
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1120..1143
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1188..1206)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1557..2360
                     /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      2443..3425
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3503..3532
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3554..4141
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4153..4180)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4204..4395)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4396..5187,5188..5256))
                     /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     5019..5038
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5257..5359)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5360..5391)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 ggaggtcagg tatgatttaa atggtcagtt agagcggccg cttgctatat tgaagtacgg
       61 attagaagcc gccgagcggg cgacagccct ccgacggaag actctcctcc gtgcgtcctc
      121 gtcttcaccg gtcgcgttcc tgaaacgcag atgtgcctcg cgccgcactg ctccgaacaa
      181 taaagattct acaatactag cttttatggt tatgaagagg aaaaattggc agtaacctgg
      241 ccccacaaac cttcaaatta acgaatcaaa ttaacaacca taggatgata atgcgattag
      301 ttttttagcc ttatttctgg ggtaattaat cagcgaagcg atgatttttg atctattaac
      361 agatatataa atggaaaagc tgcataacca ctttaactaa tactttcaac attttcagtt
      421 tgtattactt cttattcaaa tgtcataaaa gtatcaacaa aaaattgtta atatacctct
      481 atactttaac gtcaaggaga aaaaactata atgatagcct cagagtttag ttcccttcca
      541 tcatatgcag cttttgctac tgctcaagaa gcttatgagc aggctgttgc taatggtgat
      601 tctgaagttg ttcttaaaaa gttgaagaag tctttgaatg tggctaaatc tgaatttgac
      661 cgtgatgcag ccatgcaacg taagttggaa aagatggctg atcaagctat gacccaaatg
      721 tataaacagg ctagatctga ggacaagagg gcaaaagtta ctagtgctat gcagacaatg
      781 cttttcacta tgcttagaaa gttggataat gatgcactca acaacattat caacaatgca
      841 agagatggtt gtgttccctt gaacataata cctcttacaa cagcagccaa actaatggtt
      901 gtcataccag actataacac atataaaaat acgtgtgatg gtacaacatt tacttatgca
      961 tcagcattgt gggaaatcca acaggttgta gatgcagata gtaaaattgt tcaacttagt
     1021 gaaattagta tggacaattc acctaattta gcatggcctc ttattgtaac agctttaagg
     1081 gccaattctg ctgtcaaatt actcgtgccg agaggttctt ggagccatcc gcagtttgaa
     1141 aaataaacag gccccttttc ctttgtcgat atcatgtaat tagttatgtc acgcttacat
     1201 tcacgccctc ctcccacatc cgctctaacc gaaaaggaag gagttagaca acctgaagtc
     1261 taggtcccta tttatttttt ttaatagtta tgttagtatt aagaacgtta tttatatttc
     1321 aaatttttct tttttttctg tacaaagcgg ccgccgaacc actagtgcac tgcagtacac
     1381 ggtttccttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg
     1441 agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc
     1501 cagtattctt aacccaactg cacagaacaa aaacgtgcag gaaacgaaga taaatcatgt
     1561 cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct gccaagctat
     1621 ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt cgtaccacca
     1681 aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta aaaacacatg
     1741 tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag gcattatccg
     1801 ccaagtacaa ttttttactc ttcgaggaca gaaaatttgc tgacattggt aatacagtca
     1861 aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt acgaatgcac
     1921 acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa gaagtaacaa
     1981 aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc ctatctactg
     2041 gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt gttatcggct
     2101 ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg attatgacac
     2161 ccggtgtggg tttagatgac aagggagatg cattgggtca acagtataga accgtggatg
     2221 atgtggtttc tacaggatct gacattatta ttgttggaag aggactattt gcaaagggaa
     2281 gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca tatttgagaa
     2341 gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata ctaaactcac
     2401 aaattagagc ttcaatttaa ttatatcagt tattacccga gtaacgaagc atctgtgctt
     2461 cattttgtag aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc
     2521 tgcattttta cagaacagaa atgcaacgcg aaagcgctat tttaccaacg aagaatctgt
     2581 gcttcatttt tgtaaaacaa aaatgcaacg cgagagcgct aatttttcaa acaaagaatc
     2641 tgagctgcat ttttacagaa cagaaatgca acgcgagagc gctattttac caacaaagaa
     2701 tctatacttc ttttttgttc tacaaaaatg catcccgaga gcgctatttt tctaacaaag
     2761 catcttagat tacttttttt ctcctttgtg cgctctataa tgcagtctct tgataacttt
     2821 ttgcactgta ggtccgttaa ggttagaaga aggctacttt ggtgtctatt ttctcttcca
     2881 taaaaaaagc ctgactccac ttcccgcgtt tactgattac tagcgaagct gcgggtgcat
     2941 tttttcaaga taaaggcatc cccgattata ttctataccg atgtggattg cgcatacttt
     3001 gtgaacagaa agtgatagcg ttgatgattc ttcattggtc agaaaattat gaacggtttc
     3061 ttctattttg tctctatata ctacgtatag gaaatgttta cattttcgta ttgttttcga
     3121 ttcactctat gaatagttct tactacaatt tttttgtcta aagagtaata ctagagataa
     3181 acataaaaaa tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag gtggatgggt
     3241 aggttatata gggatatagc acagagatat atagcaaaga gatacttttg agcaatgttt
     3301 gtggaagcgg tattcgcaat attttagtag ctcgttacag tccggtgcgt ttttggtttt
     3361 ttgaaagtgc gtcatcagag cgcttttggt tttcaaaagc gctctgaagt tcctatactt
     3421 tctagctaga gaataggaac ttcccgagcg gccgcgtgtt acaagcttga ctccagcgta
     3481 actggactgc aatcaactca ctggctcacc ttcacgggtg ggcctttctt cggtagaaaa
     3541 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     3601 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     3661 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     3721 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3781 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3841 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3901 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     3961 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     4021 gcgcacgagg gagcttccag gggtaaacgc ctggtatctt tatagtcctg tcgggtttcg
     4081 ccacctctga cttgagcatc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     4141 aaacgccagc aacgcagaaa ggcccacccg aaggtgagcc aggtgattac atttgggccc
     4201 tcatcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc
     4261 agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag
     4321 tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt
     4381 ttcctgtttg gtcacttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4441 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tgcgggaggg
     4501 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     4561 tttatcagca ataaaccagc cagccgggag ggccgagcgc agaagtgatc ctgcaacttt
     4621 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4681 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     4741 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     4801 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgccagaa gtaagttggc
     4861 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     4921 cgtgagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     4981 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5041 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgtaaactct caaggatctt
     5101 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     5161 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     5221 gggaataagg gcgacacgga aatgttgaat actcatttta gcttccttag ctcctgaaaa
     5281 tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt gaaagttgga
     5341 acctcttacg tgccgatcaa gtcaaaagcc tccggtcgga ggcttttgac tttctgctat
//
