LOCUS       sequence-296252-        7140 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 7140)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 7140)
  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-m4046585
FEATURES             Location/Qualifiers
     source          1..7140
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        16..457
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    16..133
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     428..451
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             complement(658..669)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             1726..1743
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2886..2904)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             3255..4058
                     /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      4141..5123
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      5201..5230
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      5252..5839
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5851..5878)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5902..6093)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6094..6885,6886..6954))
                     /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     6717..6736
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6955..7057)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(7058..7089)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 tgctatattg aagtacggat tagaagccgc cgagcgggcg acagccctcc gacggaagac
       61 tctcctccgt gcgtcctcgt cttcaccggt cgcgttcctg aaacgcagat gtgcctcgcg
      121 ccgcactgct ccgaacaata aagattctac aatactagct tttatggtta tgaagaggaa
      181 aaattggcag taacctggcc ccacaaacct tcaaattaac gaatcaaatt aacaaccata
      241 ggatgataat gcgattagtt ttttagcctt atttctgggg taattaatca gcgaagcgat
      301 gatttttgat ctattaacag atatataaat ggaaaagctg cataaccact ttaactaata
      361 ctttcaacat tttcagtttg tattacttct tattcaaatg tcataaaagt atcaacaaaa
      421 aattgttaat atacctctat actttaacgt caaggagaaa aaactataat gtctgataat
      481 ggcccacaga accaacgtaa cgcacctaga atcaccttcg gtggtccatc cgacagcact
      541 ggttcaaatc aaaacggaga gagatcgggt gcccggagta agcaaaggcg cccccaaggt
      601 ctaccgaata acacagcttc ttggtttact gctcttacgc aacacggtaa agaagacttg
      661 aagttcccaa gaggtcaagg ggttccaatt aacaccaact cctctccaga tgatcaaata
      721 ggttactata gaagagctac tagacgtatt aggggtgggg acggcaaaat gaaggattta
      781 tctcctagat ggtactttta ttacttgggt accggtccag aagctggatt gccttacggt
      841 gccaataagg acggtatcat ttgggtagca actgaaggtg ctttaaatac cccaaaggac
      901 catatcggta ctagaaaccc tgctaataac gctgccattg tcttgcaatt accacagggg
      961 acaacgttgc caaagggttt ctacgccgaa ggatccagag gtggctcgca agcttctagt
     1021 agatcatcct ctcgcagcag aaactcatct cgtaactcca caccaggtag ttcgagaggt
     1081 acctctccag cgagaatggc tggtaacggt ggtgatgcag ctctggcgtt gctgctactc
     1141 gacagattga atcaattgga atccaaaatg tctggtaagg gtcaacaaca acagggtcag
     1201 actgtgacta agaagtctgc tgctgaagct tccaaaaagc cccgtcaaaa gagaaccgcc
     1261 accaaggcct ataatgtcac tcaagctttc ggaagaagag gtccagagca aacgcaaggt
     1321 aactttggag atcaagaatt gatccggcag ggtaccgatt ataaacactg gccccaaatt
     1381 gctcaattcg cgccaagcgc ctcagctttt ttcggcatgt caagaatagg tatggaagtt
     1441 actccatctg ggacttggtt aacttacact ggtgcaatca agttggacga taaagaccca
     1501 aattttaaag accaagttat tctactcaac aagcatatag atgcttacaa aaccttcccg
     1561 cctacagaac ccaagaaaga taaaaagaag aaggctgacg agacacaagc attgcctcag
     1621 cgtcaaaaga aacaacaaac tgtcaccttg ttaccagcag ctgatcttga tgatttctct
     1681 aagcaattgc aacaatccat gtcgtccgcc gactctactc aagcgctcgt gccgagaggt
     1741 tctaaaatcg aggaaggtaa gctagtcatt tggataaatg gagataaggg ttacaacggt
     1801 ttggcagaag taggcaagaa atttgaaaag gacacaggta tcaaggtgac ggttgaacat
     1861 ccagacaagt tagaagaaaa attccctcaa gttgctgcta ctggtgatgg tcccgacatt
     1921 attttctggg cccacgatcg ttttggtgga tatgctcagt caggtttgct ggctgagatc
     1981 accccagaca aggccttcca agataaattg tacccattca cttgggatgc tgtcagatac
     2041 aacggtaagc ttatagcata tccaattgct gttgaagcct tatcgttgat ctacaataag
     2101 gacttgttac ctaacccacc aaagacatgg gaagaaattc cagctttgga taaagagcta
     2161 aaggccaagg gaaaaagcgc tttgatgttt aacttacaag aaccttattt cacctggcca
     2221 ttaatcgctg ctgacggtgg ttacgcattt aaatacgaaa acggtaagta tgacattaag
     2281 gatgtcggcg ttgataatgc tggtgcaaag gctggtttga ccttcttggt tgatttaata
     2341 aagaataaac acatgaacgc tgacactgac tactctatcg ccgaagccgc tttcaacaag
     2401 ggtgaaactg ctatgaccat aaatggtcca tgggcctgga gtaacattga tacttctaaa
     2461 gtaaactatg gtgtcactgt tttgcctaca ttcaaaggcc aaccatccaa gccctttgtc
     2521 ggtgttctat ccgctggcat caatgctgct tctccaaaca aggagctagc taaagaattt
     2581 ttggaaaatt acttgttaac tgacgaaggt ttggaggccg ttaacaagga caagccattg
     2641 ggtgctgtcg ctttgaaatc ttacgaagaa gaattggcta aggatcccag aattgccgct
     2701 accatggaga acgcccaaaa gggtgaaatt atgccaaaca tcccacaaat gtcagccttc
     2761 tggtacgcag ttaggactgc cgtgattaat gctgcttccg gtagacaaac cgtcgacgaa
     2821 gcattaaaag atgctcaaac gtaaacaggc cccttttcct ttgtcgatat catgtaatta
     2881 gttatgtcac gcttacattc acgccctcct cccacatccg ctctaaccga aaaggaagga
     2941 gttagacaac ctgaagtcta ggtccctatt tatttttttt aatagttatg ttagtattaa
     3001 gaacgttatt tatatttcaa atttttcttt tttttctgta caaagcggcc gccgaaccac
     3061 tagtgcactg cagtacacgg tttccttgaa atttttttga ttcggtaatc tccgaacaga
     3121 aggaagaacg aaggaaggag cacagactta gattggtata tatacgcata tgtagtgttg
     3181 aagaaacatg aaattgccca gtattcttaa cccaactgca cagaacaaaa acgtgcagga
     3241 aacgaagata aatcatgtcg aaagctacat ataaggaacg tgctgctact catcctagtc
     3301 ctgttgctgc caagctattt aatatcatgc acgaaaagca aacaaacttg tgtgcttcat
     3361 tggatgttcg taccaccaag gaattactgg agttagttga agcattaggt cccaaaattt
     3421 gtttactaaa aacacatgtg gatatcttga ctgatttttc catggagggc acagttaagc
     3481 cgctaaaggc attatccgcc aagtacaatt ttttactctt cgaggacaga aaatttgctg
     3541 acattggtaa tacagtcaaa ttgcagtact ctgcgggtgt atacagaata gcagaatggg
     3601 cagacattac gaatgcacac ggtgtggtgg gcccaggtat tgttagcggt ttgaagcagg
     3661 cggcagaaga agtaacaaag gaacctagag gccttttgat gttagcagaa ttgtcatgca
     3721 agggctccct atctactgga gaatatacta agggtactgt tgacattgcg aagagcgaca
     3781 aagattttgt tatcggcttt attgctcaaa gagacatggg tggaagagat gaaggttacg
     3841 attggttgat tatgacaccc ggtgtgggtt tagatgacaa gggagatgca ttgggtcaac
     3901 agtatagaac cgtggatgat gtggtttcta caggatctga cattattatt gttggaagag
     3961 gactatttgc aaagggaagg gatgctaagg tagagggtga acgttacaga aaagcaggct
     4021 gggaagcata tttgagaaga tgcggccagc aaaactaaaa aactgtatta taagtaaatg
     4081 catgtatact aaactcacaa attagagctt caatttaatt atatcagtta ttacccgagt
     4141 aacgaagcat ctgtgcttca ttttgtagaa caaaaatgca acgcgagagc gctaattttt
     4201 caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgaa agcgctattt
     4261 taccaacgaa gaatctgtgc ttcatttttg taaaacaaaa atgcaacgcg agagcgctaa
     4321 tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgagagcgc
     4381 tattttacca acaaagaatc tatacttctt ttttgttcta caaaaatgca tcccgagagc
     4441 gctatttttc taacaaagca tcttagatta ctttttttct cctttgtgcg ctctataatg
     4501 cagtctcttg ataacttttt gcactgtagg tccgttaagg ttagaagaag gctactttgg
     4561 tgtctatttt ctcttccata aaaaaagcct gactccactt cccgcgttta ctgattacta
     4621 gcgaagctgc gggtgcattt tttcaagata aaggcatccc cgattatatt ctataccgat
     4681 gtggattgcg catactttgt gaacagaaag tgatagcgtt gatgattctt cattggtcag
     4741 aaaattatga acggtttctt ctattttgtc tctatatact acgtatagga aatgtttaca
     4801 ttttcgtatt gttttcgatt cactctatga atagttctta ctacaatttt tttgtctaaa
     4861 gagtaatact agagataaac ataaaaaatg tagaggtcga gtttagatgc aagttcaagg
     4921 agcgaaaggt ggatgggtag gttatatagg gatatagcac agagatatat agcaaagaga
     4981 tacttttgag caatgtttgt ggaagcggta ttcgcaatat tttagtagct cgttacagtc
     5041 cggtgcgttt ttggtttttt gaaagtgcgt catcagagcg cttttggttt tcaaaagcgc
     5101 tctgaagttc ctatactttc tagctagaga ataggaactt cccgagcggc cgcgtgttac
     5161 aagcttgact ccagcgtaac tggactgcaa tcaactcact ggctcacctt cacgggtggg
     5221 cctttcttcg gtagaaaatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     5281 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     5341 taccaactct ttttccgagg taactggctt cagcagagcg cagataccaa atactgttct
     5401 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5461 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5521 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     5581 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     5641 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     5701 cagggtcgga acaggagagc gcacgaggga gcttccaggg gtaaacgcct ggtatcttta
     5761 tagtcctgtc gggtttcgcc acctctgact tgagcatcga tttttgtgat gctcgtcagg
     5821 ggggcggagc ctatggaaaa acgccagcaa cgcagaaagg cccacccgaa ggtgagccag
     5881 gtgattacat ttgggccctc atcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca
     5941 gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc
     6001 cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc
     6061 catgttaagg gcggtttttt cctgtttggt cacttaccaa tgcttaatca gtgaggcacc
     6121 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     6181 aactacgatg cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     6241 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccgggaggg ccgagcgcag
     6301 aagtgatcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     6361 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     6421 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     6481 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     6541 tgccagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     6601 tcttactgtc atgccatccg tgagatgctt ttctgtgact ggtgagtact caaccaagtc
     6661 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     6721 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     6781 taaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     6841 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     6901 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcattttagc
     6961 ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg atcttatttc
     7021 attatggtga aagttggaac ctcttacgtg ccgatcaagt caaaagcctc cggtcggagg
     7081 cttttgactt tctgctatgg aggtcaggta tgatttaaat ggtcagttag agcggccgct
//
