LOCUS       sequence-287704-        5139 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 5139)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5139)
  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-m4046402
FEATURES             Location/Qualifiers
     source          1..5139
                     /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"
     CDS             814..831
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             832..849
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(894..912)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1263..2066
                     /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      2149..3131
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3209..3238
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3260..3847
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3859..3886)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3910..4101)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4102..4893,4894..4962))
                     /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     4725..4744
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4963..5065)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5066..5097)
                     /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 aatgagctta gtcctgttgc actacgacag atgtcttgtg ctgccggtac tacacaaact
      541 gcttgcactg atgacaatgc gttagcttac tacaacacaa caaagggagg taggtttgta
      601 cttgcactgt tatccgattt acaggatttg aaatgggcta gattccctaa gagtgatgga
      661 actggtacta tctatacaga actggaacca ccttgtaggt ttgttacaga cacacctaaa
      721 ggtcctaaag tgaagtattt atactttatt aaaggattaa acaacctaaa tagaggtatg
      781 gtacttggta gtttagctgc cacagtacgt ctactcgtgc cgagaggttc tcaccatcat
      841 caccatcatt aaacaggccc cttttccttt gtcgatatca tgtaattagt tatgtcacgc
      901 ttacattcac gccctcctcc cacatccgct ctaaccgaaa aggaaggagt tagacaacct
      961 gaagtctagg tccctattta ttttttttaa tagttatgtt agtattaaga acgttattta
     1021 tatttcaaat ttttcttttt tttctgtaca aagcggccgc cgaaccacta gtgcactgca
     1081 gtacacggtt tccttgaaat ttttttgatt cggtaatctc cgaacagaag gaagaacgaa
     1141 ggaaggagca cagacttaga ttggtatata tacgcatatg tagtgttgaa gaaacatgaa
     1201 attgcccagt attcttaacc caactgcaca gaacaaaaac gtgcaggaaa cgaagataaa
     1261 tcatgtcgaa agctacatat aaggaacgtg ctgctactca tcctagtcct gttgctgcca
     1321 agctatttaa tatcatgcac gaaaagcaaa caaacttgtg tgcttcattg gatgttcgta
     1381 ccaccaagga attactggag ttagttgaag cattaggtcc caaaatttgt ttactaaaaa
     1441 cacatgtgga tatcttgact gatttttcca tggagggcac agttaagccg ctaaaggcat
     1501 tatccgccaa gtacaatttt ttactcttcg aggacagaaa atttgctgac attggtaata
     1561 cagtcaaatt gcagtactct gcgggtgtat acagaatagc agaatgggca gacattacga
     1621 atgcacacgg tgtggtgggc ccaggtattg ttagcggttt gaagcaggcg gcagaagaag
     1681 taacaaagga acctagaggc cttttgatgt tagcagaatt gtcatgcaag ggctccctat
     1741 ctactggaga atatactaag ggtactgttg acattgcgaa gagcgacaaa gattttgtta
     1801 tcggctttat tgctcaaaga gacatgggtg gaagagatga aggttacgat tggttgatta
     1861 tgacacccgg tgtgggttta gatgacaagg gagatgcatt gggtcaacag tatagaaccg
     1921 tggatgatgt ggtttctaca ggatctgaca ttattattgt tggaagagga ctatttgcaa
     1981 agggaaggga tgctaaggta gagggtgaac gttacagaaa agcaggctgg gaagcatatt
     2041 tgagaagatg cggccagcaa aactaaaaaa ctgtattata agtaaatgca tgtatactaa
     2101 actcacaaat tagagcttca atttaattat atcagttatt acccgagtaa cgaagcatct
     2161 gtgcttcatt ttgtagaaca aaaatgcaac gcgagagcgc taatttttca aacaaagaat
     2221 ctgagctgca tttttacaga acagaaatgc aacgcgaaag cgctatttta ccaacgaaga
     2281 atctgtgctt catttttgta aaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa
     2341 agaatctgag ctgcattttt acagaacaga aatgcaacgc gagagcgcta ttttaccaac
     2401 aaagaatcta tacttctttt ttgttctaca aaaatgcatc ccgagagcgc tatttttcta
     2461 acaaagcatc ttagattact ttttttctcc tttgtgcgct ctataatgca gtctcttgat
     2521 aactttttgc actgtaggtc cgttaaggtt agaagaaggc tactttggtg tctattttct
     2581 cttccataaa aaaagcctga ctccacttcc cgcgtttact gattactagc gaagctgcgg
     2641 gtgcattttt tcaagataaa ggcatccccg attatattct ataccgatgt ggattgcgca
     2701 tactttgtga acagaaagtg atagcgttga tgattcttca ttggtcagaa aattatgaac
     2761 ggtttcttct attttgtctc tatatactac gtataggaaa tgtttacatt ttcgtattgt
     2821 tttcgattca ctctatgaat agttcttact acaatttttt tgtctaaaga gtaatactag
     2881 agataaacat aaaaaatgta gaggtcgagt ttagatgcaa gttcaaggag cgaaaggtgg
     2941 atgggtaggt tatataggga tatagcacag agatatatag caaagagata cttttgagca
     3001 atgtttgtgg aagcggtatt cgcaatattt tagtagctcg ttacagtccg gtgcgttttt
     3061 ggttttttga aagtgcgtca tcagagcgct tttggttttc aaaagcgctc tgaagttcct
     3121 atactttcta gctagagaat aggaacttcc cgagcggccg cgtgttacaa gcttgactcc
     3181 agcgtaactg gactgcaatc aactcactgg ctcaccttca cgggtgggcc tttcttcggt
     3241 agaaaatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     3301 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     3361 ttccgaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     3421 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3481 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3541 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     3601 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     3661 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     3721 aggagagcgc acgagggagc ttccaggggt aaacgcctgg tatctttata gtcctgtcgg
     3781 gtttcgccac ctctgacttg agcatcgatt tttgtgatgc tcgtcagggg ggcggagcct
     3841 atggaaaaac gccagcaacg cagaaaggcc cacccgaagg tgagccaggt gattacattt
     3901 gggccctcat cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag
     3961 ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc
     4021 gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc
     4081 ggttttttcc tgtttggtca cttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4141 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatgcg
     4201 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4261 tccagattta tcagcaataa accagccagc cgggagggcc gagcgcagaa gtgatcctgc
     4321 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4381 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     4441 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     4501 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg ccagaagtaa
     4561 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     4621 gccatccgtg agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     4681 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     4741 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgta aactctcaag
     4801 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     4861 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     4921 aaaaaaggga ataagggcga cacggaaatg ttgaatactc attttagctt ccttagctcc
     4981 tgaaaatctc gataactcaa aaaatacgcc cggtagtgat cttatttcat tatggtgaaa
     5041 gttggaacct cttacgtgcc gatcaagtca aaagcctccg gtcggaggct tttgactttc
     5101 tgctatggag gtcaggtatg atttaaatgg tcagttaga
//
