LOCUS       sequence-287673-        5193 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 5193)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5193)
  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-m4046468
FEATURES             Location/Qualifiers
     source          1..5193
                     /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             820..837
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             838..903
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(948..966)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1317..2120
                     /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      2203..3185
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3263..3292
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3314..3901
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3913..3940)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3964..4155)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4156..4947,4948..5016))
                     /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     4779..4798
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5017..5119)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5120..5151)
                     /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 aacaatgagc tttctcctgt tgctttacgg cagatgtcct gcgcagctgg aacaacgcaa
      541 actgcctgta ctgacgataa cgctctagct tactataaca ccactaaagg tggtagattc
      601 gtcttggcct tgttgtcaga cctgcaagat ttgaagtggg cgagatttcc aaagagcgat
      661 ggcactggta ccatatacac cgaattagaa ccaccctgtc gtttcgtgac cgacactcca
      721 aaaggtccga aggttaagta tttgtacttt attaagggtt taaataactt gaatcgaggt
      781 atggttctgg ggtctttggc tgctacagtc agactccaac tcgtgccgag aggttctgat
      841 tataaagatc atgatggtga ttataaagat catgatattg attataaaga tgatgatgat
      901 aaataaacag gccccttttc ctttgtcgat atcatgtaat tagttatgtc acgcttacat
      961 tcacgccctc ctcccacatc cgctctaacc gaaaaggaag gagttagaca acctgaagtc
     1021 taggtcccta tttatttttt ttaatagtta tgttagtatt aagaacgtta tttatatttc
     1081 aaatttttct tttttttctg tacaaagcgg ccgccgaacc actagtgcac tgcagtacac
     1141 ggtttccttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg
     1201 agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc
     1261 cagtattctt aacccaactg cacagaacaa aaacgtgcag gaaacgaaga taaatcatgt
     1321 cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct gccaagctat
     1381 ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt cgtaccacca
     1441 aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta aaaacacatg
     1501 tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag gcattatccg
     1561 ccaagtacaa ttttttactc ttcgaggaca gaaaatttgc tgacattggt aatacagtca
     1621 aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt acgaatgcac
     1681 acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa gaagtaacaa
     1741 aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc ctatctactg
     1801 gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt gttatcggct
     1861 ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg attatgacac
     1921 ccggtgtggg tttagatgac aagggagatg cattgggtca acagtataga accgtggatg
     1981 atgtggtttc tacaggatct gacattatta ttgttggaag aggactattt gcaaagggaa
     2041 gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca tatttgagaa
     2101 gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata ctaaactcac
     2161 aaattagagc ttcaatttaa ttatatcagt tattacccga gtaacgaagc atctgtgctt
     2221 cattttgtag aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc
     2281 tgcattttta cagaacagaa atgcaacgcg aaagcgctat tttaccaacg aagaatctgt
     2341 gcttcatttt tgtaaaacaa aaatgcaacg cgagagcgct aatttttcaa acaaagaatc
     2401 tgagctgcat ttttacagaa cagaaatgca acgcgagagc gctattttac caacaaagaa
     2461 tctatacttc ttttttgttc tacaaaaatg catcccgaga gcgctatttt tctaacaaag
     2521 catcttagat tacttttttt ctcctttgtg cgctctataa tgcagtctct tgataacttt
     2581 ttgcactgta ggtccgttaa ggttagaaga aggctacttt ggtgtctatt ttctcttcca
     2641 taaaaaaagc ctgactccac ttcccgcgtt tactgattac tagcgaagct gcgggtgcat
     2701 tttttcaaga taaaggcatc cccgattata ttctataccg atgtggattg cgcatacttt
     2761 gtgaacagaa agtgatagcg ttgatgattc ttcattggtc agaaaattat gaacggtttc
     2821 ttctattttg tctctatata ctacgtatag gaaatgttta cattttcgta ttgttttcga
     2881 ttcactctat gaatagttct tactacaatt tttttgtcta aagagtaata ctagagataa
     2941 acataaaaaa tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag gtggatgggt
     3001 aggttatata gggatatagc acagagatat atagcaaaga gatacttttg agcaatgttt
     3061 gtggaagcgg tattcgcaat attttagtag ctcgttacag tccggtgcgt ttttggtttt
     3121 ttgaaagtgc gtcatcagag cgcttttggt tttcaaaagc gctctgaagt tcctatactt
     3181 tctagctaga gaataggaac ttcccgagcg gccgcgtgtt acaagcttga ctccagcgta
     3241 actggactgc aatcaactca ctggctcacc ttcacgggtg ggcctttctt cggtagaaaa
     3301 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     3361 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     3421 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     3481 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3541 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3601 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3661 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     3721 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     3781 gcgcacgagg gagcttccag gggtaaacgc ctggtatctt tatagtcctg tcgggtttcg
     3841 ccacctctga cttgagcatc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     3901 aaacgccagc aacgcagaaa ggcccacccg aaggtgagcc aggtgattac atttgggccc
     3961 tcatcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc
     4021 agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag
     4081 tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt
     4141 ttcctgtttg gtcacttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4201 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tgcgggaggg
     4261 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     4321 tttatcagca ataaaccagc cagccgggag ggccgagcgc agaagtgatc ctgcaacttt
     4381 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4441 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     4501 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     4561 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgccagaa gtaagttggc
     4621 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     4681 cgtgagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     4741 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     4801 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgtaaactct caaggatctt
     4861 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     4921 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     4981 gggaataagg gcgacacgga aatgttgaat actcatttta gcttccttag ctcctgaaaa
     5041 tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt gaaagttgga
     5101 acctcttacg tgccgatcaa gtcaaaagcc tccggtcgga ggcttttgac tttctgctat
     5161 ggaggtcagg tatgatttaa atggtcagtt aga
//
