LOCUS       sequence-291482-        6108 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 N (nucleocapsid).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6108)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6108)
  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-m4046418
FEATURES             Location/Qualifiers
     source          1..6108
                     /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             1735..1752
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1753..1818
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(1863..1881)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2232..3035
                     /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      3118..4100
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4178..4207
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4229..4816
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4828..4855)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4879..5070)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5071..5862,5863..5931))
                     /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     5694..5713
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5932..6034)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6035..6066)
                     /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 tctgataatg gaccccaaaa tcagcgaaat gcaccccgca ttacgtttgg tggaccctca
      541 gattcaactg gcagtaacca gaatggagaa cgcagtgggg cgcgatcaaa acaacgtcgg
      601 ccccaaggtt tacccaataa tactgcgtct tggttcaccg ctctcactca acatggcaag
      661 gaagacctta aattccctcg aggacaaggc gttccaatta acaccaatag cagtccagat
      721 gaccaaattg gctactaccg aagagctacc agacgaattc gtggtggtga cggtaaaatg
      781 aaagatctca gtccaagatg gtatttctac tacctaggaa ctgggccaga agctggactt
      841 ccctatggtg ctaacaaaga cggcatcata tgggttgcaa ctgagggagc cttgaataca
      901 ccaaaagatc acattggcac ccgcaatcct gctaacaatg ctgcaatcgt gctacaactt
      961 cctcaaggaa caacattgcc aaaaggcttc tacgcagaag ggagcagagg cggcagtcaa
     1021 gcctcttctc gttcctcatc acgtagtcgc aacagttcaa gaaattcaac tccaggcagc
     1081 agtaggggaa cttctcctgc tagaatggct ggcaatggcg gtgatgctgc tcttgctttg
     1141 ctgctgcttg acagattgaa ccagcttgag agcaaaatgt ctggtaaagg ccaacaacaa
     1201 caaggccaaa ctgtcactaa gaaatctgct gctgaggctt ctaagaagcc tcggcaaaaa
     1261 cgtactgcca ctaaagcata caatgtaaca caagctttcg gcagacgtgg tccagaacaa
     1321 acccaaggaa attttgggga ccaggaacta atcagacaag gaactgatta caaacattgg
     1381 ccgcaaattg cacaatttgc ccccagcgct tcagcgttct tcggaatgtc gcgcattggc
     1441 atggaagtca caccttcggg aacgtggttg acctacacag gtgccatcaa attggatgac
     1501 aaagatccaa atttcaaaga tcaagtcatt ttgctgaata agcatattga cgcatacaaa
     1561 acattcccac caacagagcc taaaaaggac aaaaagaaga aggctgatga aactcaagcc
     1621 ttaccgcaga gacagaagaa acagcaaact gtgactcttc ttcctgctgc agatttggat
     1681 gatttctcca aacaattgca acaatccatg agcagtgctg actcaactca ggccctcgtg
     1741 ccgagaggtt ctgattataa agatcatgat ggtgattata aagatcatga tattgattat
     1801 aaagatgatg atgataaata aacaggcccc ttttcctttg tcgatatcat gtaattagtt
     1861 atgtcacgct tacattcacg ccctcctccc acatccgctc taaccgaaaa ggaaggagtt
     1921 agacaacctg aagtctaggt ccctatttat tttttttaat agttatgtta gtattaagaa
     1981 cgttatttat atttcaaatt tttctttttt ttctgtacaa agcggccgcc gaaccactag
     2041 tgcactgcag tacacggttt ccttgaaatt tttttgattc ggtaatctcc gaacagaagg
     2101 aagaacgaag gaaggagcac agacttagat tggtatatat acgcatatgt agtgttgaag
     2161 aaacatgaaa ttgcccagta ttcttaaccc aactgcacag aacaaaaacg tgcaggaaac
     2221 gaagataaat catgtcgaaa gctacatata aggaacgtgc tgctactcat cctagtcctg
     2281 ttgctgccaa gctatttaat atcatgcacg aaaagcaaac aaacttgtgt gcttcattgg
     2341 atgttcgtac caccaaggaa ttactggagt tagttgaagc attaggtccc aaaatttgtt
     2401 tactaaaaac acatgtggat atcttgactg atttttccat ggagggcaca gttaagccgc
     2461 taaaggcatt atccgccaag tacaattttt tactcttcga ggacagaaaa tttgctgaca
     2521 ttggtaatac agtcaaattg cagtactctg cgggtgtata cagaatagca gaatgggcag
     2581 acattacgaa tgcacacggt gtggtgggcc caggtattgt tagcggtttg aagcaggcgg
     2641 cagaagaagt aacaaaggaa cctagaggcc ttttgatgtt agcagaattg tcatgcaagg
     2701 gctccctatc tactggagaa tatactaagg gtactgttga cattgcgaag agcgacaaag
     2761 attttgttat cggctttatt gctcaaagag acatgggtgg aagagatgaa ggttacgatt
     2821 ggttgattat gacacccggt gtgggtttag atgacaaggg agatgcattg ggtcaacagt
     2881 atagaaccgt ggatgatgtg gtttctacag gatctgacat tattattgtt ggaagaggac
     2941 tatttgcaaa gggaagggat gctaaggtag agggtgaacg ttacagaaaa gcaggctggg
     3001 aagcatattt gagaagatgc ggccagcaaa actaaaaaac tgtattataa gtaaatgcat
     3061 gtatactaaa ctcacaaatt agagcttcaa tttaattata tcagttatta cccgagtaac
     3121 gaagcatctg tgcttcattt tgtagaacaa aaatgcaacg cgagagcgct aatttttcaa
     3181 acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgaaagc gctattttac
     3241 caacgaagaa tctgtgcttc atttttgtaa aacaaaaatg caacgcgaga gcgctaattt
     3301 ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg agagcgctat
     3361 tttaccaaca aagaatctat acttcttttt tgttctacaa aaatgcatcc cgagagcgct
     3421 atttttctaa caaagcatct tagattactt tttttctcct ttgtgcgctc tataatgcag
     3481 tctcttgata actttttgca ctgtaggtcc gttaaggtta gaagaaggct actttggtgt
     3541 ctattttctc ttccataaaa aaagcctgac tccacttccc gcgtttactg attactagcg
     3601 aagctgcggg tgcatttttt caagataaag gcatccccga ttatattcta taccgatgtg
     3661 gattgcgcat actttgtgaa cagaaagtga tagcgttgat gattcttcat tggtcagaaa
     3721 attatgaacg gtttcttcta ttttgtctct atatactacg tataggaaat gtttacattt
     3781 tcgtattgtt ttcgattcac tctatgaata gttcttacta caattttttt gtctaaagag
     3841 taatactaga gataaacata aaaaatgtag aggtcgagtt tagatgcaag ttcaaggagc
     3901 gaaaggtgga tgggtaggtt atatagggat atagcacaga gatatatagc aaagagatac
     3961 ttttgagcaa tgtttgtgga agcggtattc gcaatatttt agtagctcgt tacagtccgg
     4021 tgcgtttttg gttttttgaa agtgcgtcat cagagcgctt ttggttttca aaagcgctct
     4081 gaagttccta tactttctag ctagagaata ggaacttccc gagcggccgc gtgttacaag
     4141 cttgactcca gcgtaactgg actgcaatca actcactggc tcaccttcac gggtgggcct
     4201 ttcttcggta gaaaatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4261 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4321 caactctttt tccgaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     4381 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     4441 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     4501 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     4561 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4621 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4681 ggtcggaaca ggagagcgca cgagggagct tccaggggta aacgcctggt atctttatag
     4741 tcctgtcggg tttcgccacc tctgacttga gcatcgattt ttgtgatgct cgtcaggggg
     4801 gcggagccta tggaaaaacg ccagcaacgc agaaaggccc acccgaaggt gagccaggtg
     4861 attacatttg ggccctcatc agaggttttc accgtcatca ccgaaacgcg cgaggcagct
     4921 gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc
     4981 gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat
     5041 gttaagggcg gttttttcct gtttggtcac ttaccaatgc ttaatcagtg aggcacctat
     5101 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     5161 tacgatgcgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     5221 ctcaccggct ccagatttat cagcaataaa ccagccagcc gggagggccg agcgcagaag
     5281 tgatcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5341 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5401 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5461 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgc
     5521 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     5581 tactgtcatg ccatccgtga gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     5641 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     5701 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgtaa
     5761 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5821 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5881 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca ttttagcttc
     5941 cttagctcct gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc ttatttcatt
     6001 atggtgaaag ttggaacctc ttacgtgccg atcaagtcaa aagcctccgg tcggaggctt
     6061 ttgactttct gctatggagg tcaggtatga tttaaatggt cagttaga
//
