LOCUS       sequence-297850-        6219 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 6219)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6219)
  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-m4046309
FEATURES             Location/Qualifiers
     source          1..6219
                     /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"
     primer_bind     complement(1974..1992)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2343..3146
                     /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      3229..4211
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4289..4318
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4340..4927
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4939..4966)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4990..5181)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5182..5973,5974..6042))
                     /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     5805..5824
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6043..6145)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6146..6177)
                     /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 taaaatcgag
      841 gaaggtaagc tagtcatttg gataaatgga gataagggtt acaacggttt ggcagaagta
      901 ggcaagaaat ttgaaaagga cacaggtatc aaggtgacgg ttgaacatcc agacaagtta
      961 gaagaaaaat tccctcaagt tgctgctact ggtgatggtc ccgacattat tttctgggcc
     1021 cacgatcgtt ttggtggata tgctcagtca ggtttgctgg ctgagatcac cccagacaag
     1081 gccttccaag ataaattgta cccattcact tgggatgctg tcagatacaa cggtaagctt
     1141 atagcatatc caattgctgt tgaagcctta tcgttgatct acaataagga cttgttacct
     1201 aacccaccaa agacatggga agaaattcca gctttggata aagagctaaa ggccaaggga
     1261 aaaagcgctt tgatgtttaa cttacaagaa ccttatttca cctggccatt aatcgctgct
     1321 gacggtggtt acgcatttaa atacgaaaac ggtaagtatg acattaagga tgtcggcgtt
     1381 gataatgctg gtgcaaaggc tggtttgacc ttcttggttg atttaataaa gaataaacac
     1441 atgaacgctg acactgacta ctctatcgcc gaagccgctt tcaacaaggg tgaaactgct
     1501 atgaccataa atggtccatg ggcctggagt aacattgata cttctaaagt aaactatggt
     1561 gtcactgttt tgcctacatt caaaggccaa ccatccaagc cctttgtcgg tgttctatcc
     1621 gctggcatca atgctgcttc tccaaacaag gagctagcta aagaattttt ggaaaattac
     1681 ttgttaactg acgaaggttt ggaggccgtt aacaaggaca agccattggg tgctgtcgct
     1741 ttgaaatctt acgaagaaga attggctaag gatcccagaa ttgccgctac catggagaac
     1801 gcccaaaagg gtgaaattat gccaaacatc ccacaaatgt cagccttctg gtacgcagtt
     1861 aggactgccg tgattaatgc tgcttccggt agacaaaccg tcgacgaagc attaaaagat
     1921 gctcaaacgt aaacaggccc cttttccttt gtcgatatca tgtaattagt tatgtcacgc
     1981 ttacattcac gccctcctcc cacatccgct ctaaccgaaa aggaaggagt tagacaacct
     2041 gaagtctagg tccctattta ttttttttaa tagttatgtt agtattaaga acgttattta
     2101 tatttcaaat ttttcttttt tttctgtaca aagcggccgc cgaaccacta gtgcactgca
     2161 gtacacggtt tccttgaaat ttttttgatt cggtaatctc cgaacagaag gaagaacgaa
     2221 ggaaggagca cagacttaga ttggtatata tacgcatatg tagtgttgaa gaaacatgaa
     2281 attgcccagt attcttaacc caactgcaca gaacaaaaac gtgcaggaaa cgaagataaa
     2341 tcatgtcgaa agctacatat aaggaacgtg ctgctactca tcctagtcct gttgctgcca
     2401 agctatttaa tatcatgcac gaaaagcaaa caaacttgtg tgcttcattg gatgttcgta
     2461 ccaccaagga attactggag ttagttgaag cattaggtcc caaaatttgt ttactaaaaa
     2521 cacatgtgga tatcttgact gatttttcca tggagggcac agttaagccg ctaaaggcat
     2581 tatccgccaa gtacaatttt ttactcttcg aggacagaaa atttgctgac attggtaata
     2641 cagtcaaatt gcagtactct gcgggtgtat acagaatagc agaatgggca gacattacga
     2701 atgcacacgg tgtggtgggc ccaggtattg ttagcggttt gaagcaggcg gcagaagaag
     2761 taacaaagga acctagaggc cttttgatgt tagcagaatt gtcatgcaag ggctccctat
     2821 ctactggaga atatactaag ggtactgttg acattgcgaa gagcgacaaa gattttgtta
     2881 tcggctttat tgctcaaaga gacatgggtg gaagagatga aggttacgat tggttgatta
     2941 tgacacccgg tgtgggttta gatgacaagg gagatgcatt gggtcaacag tatagaaccg
     3001 tggatgatgt ggtttctaca ggatctgaca ttattattgt tggaagagga ctatttgcaa
     3061 agggaaggga tgctaaggta gagggtgaac gttacagaaa agcaggctgg gaagcatatt
     3121 tgagaagatg cggccagcaa aactaaaaaa ctgtattata agtaaatgca tgtatactaa
     3181 actcacaaat tagagcttca atttaattat atcagttatt acccgagtaa cgaagcatct
     3241 gtgcttcatt ttgtagaaca aaaatgcaac gcgagagcgc taatttttca aacaaagaat
     3301 ctgagctgca tttttacaga acagaaatgc aacgcgaaag cgctatttta ccaacgaaga
     3361 atctgtgctt catttttgta aaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa
     3421 agaatctgag ctgcattttt acagaacaga aatgcaacgc gagagcgcta ttttaccaac
     3481 aaagaatcta tacttctttt ttgttctaca aaaatgcatc ccgagagcgc tatttttcta
     3541 acaaagcatc ttagattact ttttttctcc tttgtgcgct ctataatgca gtctcttgat
     3601 aactttttgc actgtaggtc cgttaaggtt agaagaaggc tactttggtg tctattttct
     3661 cttccataaa aaaagcctga ctccacttcc cgcgtttact gattactagc gaagctgcgg
     3721 gtgcattttt tcaagataaa ggcatccccg attatattct ataccgatgt ggattgcgca
     3781 tactttgtga acagaaagtg atagcgttga tgattcttca ttggtcagaa aattatgaac
     3841 ggtttcttct attttgtctc tatatactac gtataggaaa tgtttacatt ttcgtattgt
     3901 tttcgattca ctctatgaat agttcttact acaatttttt tgtctaaaga gtaatactag
     3961 agataaacat aaaaaatgta gaggtcgagt ttagatgcaa gttcaaggag cgaaaggtgg
     4021 atgggtaggt tatataggga tatagcacag agatatatag caaagagata cttttgagca
     4081 atgtttgtgg aagcggtatt cgcaatattt tagtagctcg ttacagtccg gtgcgttttt
     4141 ggttttttga aagtgcgtca tcagagcgct tttggttttc aaaagcgctc tgaagttcct
     4201 atactttcta gctagagaat aggaacttcc cgagcggccg cgtgttacaa gcttgactcc
     4261 agcgtaactg gactgcaatc aactcactgg ctcaccttca cgggtgggcc tttcttcggt
     4321 agaaaatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     4381 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     4441 ttccgaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     4501 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     4561 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     4621 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     4681 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     4741 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     4801 aggagagcgc acgagggagc ttccaggggt aaacgcctgg tatctttata gtcctgtcgg
     4861 gtttcgccac ctctgacttg agcatcgatt tttgtgatgc tcgtcagggg ggcggagcct
     4921 atggaaaaac gccagcaacg cagaaaggcc cacccgaagg tgagccaggt gattacattt
     4981 gggccctcat cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag
     5041 ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc
     5101 gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc
     5161 ggttttttcc tgtttggtca cttaccaatg cttaatcagt gaggcaccta tctcagcgat
     5221 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatgcg
     5281 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     5341 tccagattta tcagcaataa accagccagc cgggagggcc gagcgcagaa gtgatcctgc
     5401 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     5461 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     5521 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5581 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg ccagaagtaa
     5641 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5701 gccatccgtg agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5761 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5821 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgta aactctcaag
     5881 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5941 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     6001 aaaaaaggga ataagggcga cacggaaatg ttgaatactc attttagctt ccttagctcc
     6061 tgaaaatctc gataactcaa aaaatacgcc cggtagtgat cttatttcat tatggtgaaa
     6121 gttggaacct cttacgtgcc gatcaagtca aaagcctccg gtcggaggct tttgactttc
     6181 tgctatggag gtcaggtatg atttaaatgg tcagttaga
//
