LOCUS       sequence-298292-        5358 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 5358)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5358)
  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-m4046643
FEATURES             Location/Qualifiers
     source          1..5358
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        25..466
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    25..142
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     437..460
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     complement(1113..1131)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1482..2285
                     /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      2368..3350
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3428..3457
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3479..4066
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4078..4105)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4129..4320)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4321..5112,5113..5181))
                     /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     4944..4963
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5182..5284)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5285..5316)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgctt gctatattga agtacggatt agaagccgcc gagcgggcga cagccctccg
       61 acggaagact ctcctccgtg cgtcctcgtc ttcaccggtc gcgttcctga aacgcagatg
      121 tgcctcgcgc cgcactgctc cgaacaataa agattctaca atactagctt ttatggttat
      181 gaagaggaaa aattggcagt aacctggccc cacaaacctt caaattaacg aatcaaatta
      241 acaaccatag gatgataatg cgattagttt tttagcctta tttctggggt aattaatcag
      301 cgaagcgatg atttttgatc tattaacaga tatataaatg gaaaagctgc ataaccactt
      361 taactaatac tttcaacatt ttcagtttgt attacttctt attcaaatgt cataaaagta
      421 tcaacaaaaa attgttaata tacctctata ctttaacgtc aaggagaaaa aactataatg
      481 atagcctcag agtttagttc ccttccatca tatgcagctt ttgctactgc tcaagaagct
      541 tatgagcagg ctgttgctaa tggtgattct gaagttgttc ttaaaaagtt gaagaagtct
      601 ttgaatgtgg ctaaatctga atttgaccgt gatgcagcca tgcaacgtaa gttggaaaag
      661 atggctgatc aagctatgac ccaaatgtat aaacaggcta gatctgagga caagagggca
      721 aaagttacta gtgctatgca gacaatgctt ttcactatgc ttagaaagtt ggataatgat
      781 gcactcaaca acattatcaa caatgcaaga gatggttgtg ttcccttgaa cataatacct
      841 cttacaacag cagccaaact aatggttgtc ataccagact ataacacata taaaaatacg
      901 tgtgatggta caacatttac ttatgcatca gcattgtggg aaatccaaca ggttgtagat
      961 gcagatagta aaattgttca acttagtgaa attagtatgg acaattcacc taatttagca
     1021 tggcctctta ttgtaacagc tttaagggcc aattctgctg tcaaattata aacaggcccc
     1081 ttttcctttg tcgatatcat gtaattagtt atgtcacgct tacattcacg ccctcctccc
     1141 acatccgctc taaccgaaaa ggaaggagtt agacaacctg aagtctaggt ccctatttat
     1201 tttttttaat agttatgtta gtattaagaa cgttatttat atttcaaatt tttctttttt
     1261 ttctgtacaa agcggccgcc gaaccactag tgcactgcag tacacggttt ccttgaaatt
     1321 tttttgattc ggtaatctcc gaacagaagg aagaacgaag gaaggagcac agacttagat
     1381 tggtatatat acgcatatgt agtgttgaag aaacatgaaa ttgcccagta ttcttaaccc
     1441 aactgcacag aacaaaaacg tgcaggaaac gaagataaat catgtcgaaa gctacatata
     1501 aggaacgtgc tgctactcat cctagtcctg ttgctgccaa gctatttaat atcatgcacg
     1561 aaaagcaaac aaacttgtgt gcttcattgg atgttcgtac caccaaggaa ttactggagt
     1621 tagttgaagc attaggtccc aaaatttgtt tactaaaaac acatgtggat atcttgactg
     1681 atttttccat ggagggcaca gttaagccgc taaaggcatt atccgccaag tacaattttt
     1741 tactcttcga ggacagaaaa tttgctgaca ttggtaatac agtcaaattg cagtactctg
     1801 cgggtgtata cagaatagca gaatgggcag acattacgaa tgcacacggt gtggtgggcc
     1861 caggtattgt tagcggtttg aagcaggcgg cagaagaagt aacaaaggaa cctagaggcc
     1921 ttttgatgtt agcagaattg tcatgcaagg gctccctatc tactggagaa tatactaagg
     1981 gtactgttga cattgcgaag agcgacaaag attttgttat cggctttatt gctcaaagag
     2041 acatgggtgg aagagatgaa ggttacgatt ggttgattat gacacccggt gtgggtttag
     2101 atgacaaggg agatgcattg ggtcaacagt atagaaccgt ggatgatgtg gtttctacag
     2161 gatctgacat tattattgtt ggaagaggac tatttgcaaa gggaagggat gctaaggtag
     2221 agggtgaacg ttacagaaaa gcaggctggg aagcatattt gagaagatgc ggccagcaaa
     2281 actaaaaaac tgtattataa gtaaatgcat gtatactaaa ctcacaaatt agagcttcaa
     2341 tttaattata tcagttatta cccgagtaac gaagcatctg tgcttcattt tgtagaacaa
     2401 aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa
     2461 cagaaatgca acgcgaaagc gctattttac caacgaagaa tctgtgcttc atttttgtaa
     2521 aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta
     2581 cagaacagaa atgcaacgcg agagcgctat tttaccaaca aagaatctat acttcttttt
     2641 tgttctacaa aaatgcatcc cgagagcgct atttttctaa caaagcatct tagattactt
     2701 tttttctcct ttgtgcgctc tataatgcag tctcttgata actttttgca ctgtaggtcc
     2761 gttaaggtta gaagaaggct actttggtgt ctattttctc ttccataaaa aaagccnnnn
     2821 nncacttccc gcgtttactg attactagcg aagctgcggg tgcatttttt caagataaag
     2881 gcatccccga ttatattcta taccgatgtg gattgcgcat actttgtgaa cagaaagtga
     2941 tagcgttgat gattcttcat tggtcagaaa attatgaacg gtttcttcta ttttgtctct
     3001 atatactacg tataggaaat gtttacattt tcgtattgtt ttcgattcac tctatgaata
     3061 gttcttacta caattttttt gtctaaagag taatactaga gataaacata aaaaatgtag
     3121 aggtcgagtt tagatgcaag ttcaaggagc gaaaggtgga tgggtaggtt atatagggat
     3181 atagcacaga gatatatagc aaagagatac ttttgagcaa tgtttgtgga agcggtattc
     3241 gcaatatttt agtagctcgt tacagtccgg tgcgtttttg gttttttgaa agtgcgtcat
     3301 cagagcgctt ttggttttca aaagcgctct gaagttccta tactttctag ctagagaata
     3361 ggaacttccc gagcggccgc gtgttacaag cttgactcca gcgtaactgg actgcaatca
     3421 actcactggc tcaccttcac gggtgggcct ttcttcggta gaaaatcaaa ggatcttctt
     3481 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     3541 cggtggtttg tttgccggat caagagctac caactctttt tccgaggtaa ctggcttcag
     3601 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     3661 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     3721 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     3781 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     3841 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     3901 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     3961 tccaggggta aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4021 gcatcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     4081 agaaaggccc acccgaaggt gagccaggtg attacatttg ggccctcatc agaggttttc
     4141 accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag
     4201 cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt
     4261 taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac
     4321 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4381 agttgcctga ctccccgtcg tgtagataac tacgatgcgg gagggcttac catctggccc
     4441 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     4501 ccagccagcc gggagggccg agcgcagaag tgatcctgca actttatccg cctccatcca
     4561 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4621 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4681 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4741 ggttagctcc ttcggtcctc cgatcgttgc cagaagtaag ttggccgcag tgttatcact
     4801 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtga gatgcttttc
     4861 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     4921 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     4981 catcattgga aaacgttctt cggggcgtaa actctcaagg atcttaccgc tgttgagatc
     5041 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     5101 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     5161 acggaaatgt tgaatactca ttttagcttc cttagctcct gaaaatctcg ataactcaaa
     5221 aaatacgccc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc ttacgtgccg
     5281 atcaagtcaa aagcctccgg tcggaggctt ttgactttct gctatggagg tcaggtatga
     5341 tttaaatggt cagttaga
//
