LOCUS       sequence-298245-        5103 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 5103)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5103)
  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-m4046635
FEATURES             Location/Qualifiers
     source          1..5103
                     /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(858..876)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1227..2030
                     /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      2113..3095
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3173..3202
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3224..3811
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3823..3850)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3874..4065)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4066..4857,4858..4926))
                     /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     4689..4708
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4927..5029)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5030..5061)
                     /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 ctataaacag gccccttttc ctttgtcgat
      841 atcatgtaat tagttatgtc acgcttacat tcacgccctc ctcccacatc cgctctaacc
      901 gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt ttaatagtta
      961 tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg tacaaagcgg
     1021 ccgccgaacc actagtgcac tgcagtacac ggtttccttg aaattttttt gattcggtaa
     1081 tctccgaaca gaaggaagaa cgaaggaagg agcacagact tagattggta tatatacgca
     1141 tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt aacccaactg cacagaacaa
     1201 aaacgtgcag gaaacgaaga taaatcatgt cgaaagctac atataaggaa cgtgctgcta
     1261 ctcatcctag tcctgttgct gccaagctat ttaatatcat gcacgaaaag caaacaaact
     1321 tgtgtgcttc attggatgtt cgtaccacca aggaattact ggagttagtt gaagcattag
     1381 gtcccaaaat ttgtttacta aaaacacatg tggatatctt gactgatttt tccatggagg
     1441 gcacagttaa gccgctaaag gcattatccg ccaagtacaa ttttttactc ttcgaggaca
     1501 gaaaatttgc tgacattggt aatacagtca aattgcagta ctctgcgggt gtatacagaa
     1561 tagcagaatg ggcagacatt acgaatgcac acggtgtggt gggcccaggt attgttagcg
     1621 gtttgaagca ggcggcagaa gaagtaacaa aggaacctag aggccttttg atgttagcag
     1681 aattgtcatg caagggctcc ctatctactg gagaatatac taagggtact gttgacattg
     1741 cgaagagcga caaagatttt gttatcggct ttattgctca aagagacatg ggtggaagag
     1801 atgaaggtta cgattggttg attatgacac ccggtgtggg tttagatgac aagggagatg
     1861 cattgggtca acagtataga accgtggatg atgtggtttc tacaggatct gacattatta
     1921 ttgttggaag aggactattt gcaaagggaa gggatgctaa ggtagagggt gaacgttaca
     1981 gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa aaaactgtat
     2041 tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa ttatatcagt
     2101 tattacccga gtaacgaagc atctgtgctt cattttgtag aacaaaaatg caacgcgaga
     2161 gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg
     2221 aaagcgctat tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa aaatgcaacg
     2281 cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa cagaaatgca
     2341 acgcgagagc gctattttac caacaaagaa tctatacttc ttttttgttc tacaaaaatg
     2401 catcccgaga gcgctatttt tctaacaaag catcttagat tacttttttt ctcctttgtg
     2461 cgctctataa tgcagtctct tgataacttt ttgcactgta ggtccgttaa ggttagaaga
     2521 aggctacttt ggtgtctatt ttctcttcca taaaaaaagc ctgactccac ttcccgcgtt
     2581 tactgattac tagcgaagct gcgggtgcat tttttcaaga taaaggcatc cccgattata
     2641 ttctataccg atgtggattg cgcatacttt gtgaacagaa agtgatagcg ttgatgattc
     2701 ttcattggtc agaaaattat gaacggtttc ttctattttg tctctatata ctacgtatag
     2761 gaaatgttta cattttcgta ttgttttcga ttcactctat gaatagttct tactacaatt
     2821 tttttgtcta aagagtaata ctagagataa acataaaaaa tgtagaggtc gagtttagat
     2881 gcaagttcaa ggagcgaaag gtggatgggt aggttatata gggatatagc acagagatat
     2941 atagcaaaga gatacttttg agcaatgttt gtggaagcgg tattcgcaat attttagtag
     3001 ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc gtcatcagag cgcttttggt
     3061 tttcaaaagc gctctgaagt tcctatactt tctagctaga gaataggaac ttcccgagcg
     3121 gccgcgtgtt acaagcttga ctccagcgta actggactgc aatcaactca ctggctcacc
     3181 ttcacgggtg ggcctttctt cggtagaaaa tcaaaggatc ttcttgagat cctttttttc
     3241 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     3301 cggatcaaga gctaccaact ctttttccga ggtaactggc ttcagcagag cgcagatacc
     3361 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3421 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3481 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3541 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3601 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3661 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag gggtaaacgc
     3721 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcatc gatttttgtg
     3781 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcagaaa ggcccacccg
     3841 aaggtgagcc aggtgattac atttgggccc tcatcagagg ttttcaccgt catcaccgaa
     3901 acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc
     3961 tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct
     4021 gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcacttacc aatgcttaat
     4081 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     4141 cgtcgtgtag ataactacga tgcgggaggg cttaccatct ggccccagtg ctgcaatgat
     4201 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccgggag
     4261 ggccgagcgc agaagtgatc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     4321 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     4381 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     4441 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     4501 tcctccgatc gttgccagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     4561 actgcataat tctcttactg tcatgccatc cgtgagatgc ttttctgtga ctggtgagta
     4621 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     4681 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     4741 ttcttcgggg cgtaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     4801 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     4861 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     4921 actcatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag
     4981 tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa gtcaaaagcc
     5041 tccggtcgga ggcttttgac tttctgctat ggaggtcagg tatgatttaa atggtcagtt
     5101 aga
//
