LOCUS       sequence-292521-        5145 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 nsp9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5145)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5145)
  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-m4046347
FEATURES             Location/Qualifiers
     source          1..5145
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        48..489
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    48..165
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     460..483
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             837..854
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             855..878
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(923..941)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1292..2095
                     /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      2178..3160
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3238..3267
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3289..3876
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3888..3915)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3939..4130)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4131..4922,4923..4991))
                     /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     4754..4773
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4992..5094)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5095..5126)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 tatgatttaa atggtcagtt agagcggccg cttgctatat tgaagtacgg attagaagcc
       61 gccgagcggg cgacagccct ccgacggaag actctcctcc gtgcgtcctc gtcttcaccg
      121 gtcgcgttcc tgaaacgcag atgtgcctcg cgccgcactg ctccgaacaa taaagattct
      181 acaatactag cttttatggt tatgaagagg aaaaattggc agtaacctgg ccccacaaac
      241 cttcaaatta acgaatcaaa ttaacaacca taggatgata atgcgattag ttttttagcc
      301 ttatttctgg ggtaattaat cagcgaagcg atgatttttg atctattaac agatatataa
      361 atggaaaagc tgcataacca ctttaactaa tactttcaac attttcagtt tgtattactt
      421 cttattcaaa tgtcataaaa gtatcaacaa aaaattgtta atatacctct atactttaac
      481 gtcaaggaga aaaaactata atgaatgagc ttagtcctgt tgcactacga cagatgtctt
      541 gtgctgccgg tactacacaa actgcttgca ctgatgacaa tgcgttagct tactacaaca
      601 caacaaaggg aggtaggttt gtacttgcac tgttatccga tttacaggat ttgaaatggg
      661 ctagattccc taagagtgat ggaactggta ctatctatac agaactggaa ccaccttgta
      721 ggtttgttac agacacacct aaaggtccta aagtgaagta tttatacttt attaaaggat
      781 taaacaacct aaatagaggt atggtacttg gtagtttagc tgccacagta cgtctactcg
      841 tgccgagagg ttcttggagc catccgcagt ttgaaaaata aacaggcccc ttttcctttg
      901 tcgatatcat gtaattagtt atgtcacgct tacattcacg ccctcctccc acatccgctc
      961 taaccgaaaa ggaaggagtt agacaacctg aagtctaggt ccctatttat tttttttaat
     1021 agttatgtta gtattaagaa cgttatttat atttcaaatt tttctttttt ttctgtacaa
     1081 agcggccgcc gaaccactag tgcactgcag tacacggttt ccttgaaatt tttttgattc
     1141 ggtaatctcc gaacagaagg aagaacgaag gaaggagcac agacttagat tggtatatat
     1201 acgcatatgt agtgttgaag aaacatgaaa ttgcccagta ttcttaaccc aactgcacag
     1261 aacaaaaacg tgcaggaaac gaagataaat catgtcgaaa gctacatata aggaacgtgc
     1321 tgctactcat cctagtcctg ttgctgccaa gctatttaat atcatgcacg aaaagcaaac
     1381 aaacttgtgt gcttcattgg atgttcgtac caccaaggaa ttactggagt tagttgaagc
     1441 attaggtccc aaaatttgtt tactaaaaac acatgtggat atcttgactg atttttccat
     1501 ggagggcaca gttaagccgc taaaggcatt atccgccaag tacaattttt tactcttcga
     1561 ggacagaaaa tttgctgaca ttggtaatac agtcaaattg cagtactctg cgggtgtata
     1621 cagaatagca gaatgggcag acattacgaa tgcacacggt gtggtgggcc caggtattgt
     1681 tagcggtttg aagcaggcgg cagaagaagt aacaaaggaa cctagaggcc ttttgatgtt
     1741 agcagaattg tcatgcaagg gctccctatc tactggagaa tatactaagg gtactgttga
     1801 cattgcgaag agcgacaaag attttgttat cggctttatt gctcaaagag acatgggtgg
     1861 aagagatgaa ggttacgatt ggttgattat gacacccggt gtgggtttag atgacaaggg
     1921 agatgcattg ggtcaacagt atagaaccgt ggatgatgtg gtttctacag gatctgacat
     1981 tattattgtt ggaagaggac tatttgcaaa gggaagggat gctaaggtag agggtgaacg
     2041 ttacagaaaa gcaggctggg aagcatattt gagaagatgc ggccagcaaa actaaaaaac
     2101 tgtattataa gtaaatgcat gtatactaaa ctcacaaatt agagcttcaa tttaattata
     2161 tcagttatta cccgagtaac gaagcatctg tgcttcattt tgtagaacaa aaatgcaacg
     2221 cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa cagaaatgca
     2281 acgcgaaagc gctattttac caacgaagaa tctgtgcttc atttttgtaa aacaaaaatg
     2341 caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa
     2401 atgcaacgcg agagcgctat tttaccaaca aagaatctat acttcttttt tgttctacaa
     2461 aaatgcatcc cgagagcgct atttttctaa caaagcatct tagattactt tttttctcct
     2521 ttgtgcgctc tataatgcag tctcttgata actttttgca ctgtaggtcc gttaaggtta
     2581 gaagaaggct actttggtgt ctattttctc ttccataaaa aaagcctgac tccacttccc
     2641 gcgtttactg attactagcg aagctgcggg tgcatttttt caagataaag gcatccccga
     2701 ttatattcta taccgatgtg gattgcgcat actttgtgaa cagaaagtga tagcgttgat
     2761 gattcttcat tggtcagaaa attatgaacg gtttcttcta ttttgtctct atatactacg
     2821 tataggaaat gtttacattt tcgtattgtt ttcgattcac tctatgaata gttcttacta
     2881 caattttttt gtctaaagag taatactaga gataaacata aaaaatgtag aggtcgagtt
     2941 tagatgcaag ttcaaggagc gaaaggtgga tgggtaggtt atatagggat atagcacaga
     3001 gatatatagc aaagagatac ttttgagcaa tgtttgtgga agcggtattc gcaatatttt
     3061 agtagctcgt tacagtccgg tgcgtttttg gttttttgaa agtgcgtcat cagagcgctt
     3121 ttggttttca aaagcgctct gaagttccta tactttctag ctagagaata ggaacttccc
     3181 gagcggccgc gtgttacaag cttgactcca gcgtaactgg actgcaatca actcactggc
     3241 tcaccttcac gggtgggcct ttcttcggta gaaaatcaaa ggatcttctt gagatccttt
     3301 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     3361 tttgccggat caagagctac caactctttt tccgaggtaa ctggcttcag cagagcgcag
     3421 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     3481 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     3541 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     3601 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     3661 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     3721 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggta
     3781 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcatcgattt
     3841 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc agaaaggccc
     3901 acccgaaggt gagccaggtg attacatttg ggccctcatc agaggttttc accgtcatca
     3961 ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag
     4021 atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg
     4081 cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac ttaccaatgc
     4141 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     4201 ctccccgtcg tgtagataac tacgatgcgg gagggcttac catctggccc cagtgctgca
     4261 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     4321 gggagggccg agcgcagaag tgatcctgca actttatccg cctccatcca gtctattaat
     4381 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     4441 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     4501 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     4561 ttcggtcctc cgatcgttgc cagaagtaag ttggccgcag tgttatcact catggttatg
     4621 gcagcactgc ataattctct tactgtcatg ccatccgtga gatgcttttc tgtgactggt
     4681 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     4741 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     4801 aaacgttctt cggggcgtaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     4861 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     4921 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     4981 tgaatactca ttttagcttc cttagctcct gaaaatctcg ataactcaaa aaatacgccc
     5041 ggtagtgatc ttatttcatt atggtgaaag ttggaacctc ttacgtgccg atcaagtcaa
     5101 aagcctccgg tcggaggctt ttgactttct gctatggagg tcagg
//
