LOCUS       sequence-298147-        5752 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 N (nucleocapsid).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5752)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5752)
  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-m4046173
FEATURES             Location/Qualifiers
     source          1..5752
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(311..322)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             1379..1396
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1397..1462
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(1507..1525)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1876..2679
                     /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      2762..3744
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3822..3851
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3873..4460
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4472..4499)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4523..4714)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4715..5506,5507..5575))
                     /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     5338..5357
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5576..5678)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5679..5710)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 taataaacgg tcttcaattt ctcaagtttc agtttcattt ttcttgttct attacaactt
       61 tttttacttc ttgctcatta gaaagaaagc atagcaatct aatctaagtt ttaattacaa
      121 aatgtctgat aatggcccac agaaccaacg taacgcacct agaatcacct tcggtggtcc
      181 atccgacagc actggttcaa atcaaaacgg agagagatcg ggtgcccgga gtaagcaaag
      241 gcgcccccaa ggtctaccga ataacacagc ttcttggttt actgctctta cgcaacacgg
      301 taaagaagac ttgaagttcc caagaggtca aggggttcca attaacacca actcctctcc
      361 agatgatcaa ataggttact atagaagagc tactagacgt attaggggtg gggacggcaa
      421 aatgaaggat ttatctccta gatggtactt ttattacttg ggtaccggtc cagaagctgg
      481 attgccttac ggtgccaata aggacggtat catttgggta gcaactgaag gtgctttaaa
      541 taccccaaag gaccatatcg gtactagaaa ccctgctaat aacgctgcca ttgtcttgca
      601 attaccacag gggacaacgt tgccaaaggg tttctacgcc gaaggatcca gaggtggctc
      661 gcaagcttct agtagatcat cctctcgcag cagaaactca tctcgtaact ccacaccagg
      721 tagttcgaga ggtacctctc cagcgagaat ggctggtaac ggtggtgatg cagctctggc
      781 gttgctgcta ctcgacagat tgaatcaatt ggaatccaaa atgtctggta agggtcaaca
      841 acaacagggt cagactgtga ctaagaagtc tgctgctgaa gcttccaaaa agccccgtca
      901 aaagagaacc gccaccaagg cctataatgt cactcaagct ttcggaagaa gaggtccaga
      961 gcaaacgcaa ggtaactttg gagatcaaga attgatccgg cagggtaccg attataaaca
     1021 ctggccccaa attgctcaat tcgcgccaag cgcctcagct tttttcggca tgtcaagaat
     1081 aggtatggaa gttactccat ctgggacttg gttaacttac actggtgcaa tcaagttgga
     1141 cgataaagac ccaaatttta aagaccaagt tattctactc aacaagcata tagatgctta
     1201 caaaaccttc ccgcctacag aacccaagaa agataaaaag aagaaggctg acgagacaca
     1261 agcattgcct cagcgtcaaa agaaacaaca aactgtcacc ttgttaccag cagctgatct
     1321 tgatgatttc tctaagcaat tgcaacaatc catgtcgtcc gccgactcta ctcaagcgct
     1381 cgtgccgaga ggttctgatt ataaagatca tgatggtgat tataaagatc atgatattga
     1441 ttataaagat gatgatgata aataaacagg ccccttttcc tttgtcgata tcatgtaatt
     1501 agttatgtca cgcttacatt cacgccctcc tcccacatcc gctctaaccg aaaaggaagg
     1561 agttagacaa cctgaagtct aggtccctat ttattttttt taatagttat gttagtatta
     1621 agaacgttat ttatatttca aatttttctt ttttttctgt acaaagcggc cgccgaacca
     1681 ctagtgcact gcagtacacg gtttccttga aatttttttg attcggtaat ctccgaacag
     1741 aaggaagaac gaaggaagga gcacagactt agattggtat atatacgcat atgtagtgtt
     1801 gaagaaacat gaaattgccc agtattctta acccaactgc acagaacaaa aacgtgcagg
     1861 aaacgaagat aaatcatgtc gaaagctaca tataaggaac gtgctgctac tcatcctagt
     1921 cctgttgctg ccaagctatt taatatcatg cacgaaaagc aaacaaactt gtgtgcttca
     1981 ttggatgttc gtaccaccaa ggaattactg gagttagttg aagcattagg tcccaaaatt
     2041 tgtttactaa aaacacatgt ggatatcttg actgattttt ccatggaggg cacagttaag
     2101 ccgctaaagg cattatccgc caagtacaat tttttactct tcgaggacag aaaatttgct
     2161 gacattggta atacagtcaa attgcagtac tctgcgggtg tatacagaat agcagaatgg
     2221 gcagacatta cgaatgcaca cggtgtggtg ggcccaggta ttgttagcgg tttgaagcag
     2281 gcggcagaag aagtaacaaa ggaacctaga ggccttttga tgttagcaga attgtcatgc
     2341 aagggctccc tatctactgg agaatatact aagggtactg ttgacattgc gaagagcgac
     2401 aaagattttg ttatcggctt tattgctcaa agagacatgg gtggaagaga tgaaggttac
     2461 gattggttga ttatgacacc cggtgtgggt ttagatgaca agggagatgc attgggtcaa
     2521 cagtatagaa ccgtggatga tgtggtttct acaggatctg acattattat tgttggaaga
     2581 ggactatttg caaagggaag ggatgctaag gtagagggtg aacgttacag aaaagcaggc
     2641 tgggaagcat atttgagaag atgcggccag caaaactaaa aaactgtatt ataagtaaat
     2701 gcatgtatac taaactcaca aattagagct tcaatttaat tatatcagtt attacccgag
     2761 taacgaagca tctgtgcttc attttgtaga acaaaaatgc aacgcgagag cgctaatttt
     2821 tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga aagcgctatt
     2881 ttaccaacga agaatctgtg cttcattttt gtaaaacaaa aatgcaacgc gagagcgcta
     2941 atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgagagcg
     3001 ctattttacc aacaaagaat ctatacttct tttttgttct acaaaaatgc atcccgagag
     3061 cgctattttt ctaacaaagc atcttagatt actttttttc tcctttgtgc gctctataat
     3121 gcagtctctt gataactttt tgcactgtag gtccgttaag gttagaagaa ggctactttg
     3181 gtgtctattt tctcttccat aaaaaaagcc tgactccact tcccgcgttt actgattact
     3241 agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc ccgattatat tctataccga
     3301 tgtggattgc gcatactttg tgaacagaaa gtgatagcgt tgatgattct tcattggtca
     3361 gaaaattatg aacggtttct tctattttgt ctctatatac tacgtatagg aaatgtttac
     3421 attttcgtat tgttttcgat tcactctatg aatagttctt actacaattt ttttgtctaa
     3481 agagtaatac tagagataaa cataaaaaat gtagaggtcg agtttagatg caagttcaag
     3541 gagcgaaagg tggatgggta ggttatatag ggatatagca cagagatata tagcaaagag
     3601 atacttttga gcaatgtttg tggaagcggt attcgcaata ttttagtagc tcgttacagt
     3661 ccggtgcgtt tttggttttt tgaaagtgcg tcatcagagc gcttttggtt ttcaaaagcg
     3721 ctctgaagtt cctatacttt ctagctagag aataggaact tcccgagcgg ccgcgtgtta
     3781 caagcttgac tccagcgtaa ctggactgca atcaactcac tggctcacct tcacgggtgg
     3841 gcctttcttc ggtagaaaat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     3901 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     3961 ctaccaactc tttttccgag gtaactggct tcagcagagc gcagatacca aatactgttc
     4021 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     4081 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     4141 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     4201 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     4261 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     4321 gcagggtcgg aacaggagag cgcacgaggg agcttccagg ggtaaacgcc tggtatcttt
     4381 atagtcctgt cgggtttcgc cacctctgac ttgagcatcg atttttgtga tgctcgtcag
     4441 gggggcggag cctatggaaa aacgccagca acgcagaaag gcccacccga aggtgagcca
     4501 ggtgattaca tttgggccct catcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc
     4561 agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat
     4621 ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg
     4681 ccatgttaag ggcggttttt tcctgtttgg tcacttacca atgcttaatc agtgaggcac
     4741 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     4801 taactacgat gcgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     4861 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccgggagg gccgagcgca
     4921 gaagtgatcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     4981 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     5041 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     5101 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     5161 ttgccagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     5221 ctcttactgt catgccatcc gtgagatgct tttctgtgac tggtgagtac tcaaccaagt
     5281 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     5341 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     5401 gtaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     5461 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     5521 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcattttag
     5581 cttccttagc tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt
     5641 cattatggtg aaagttggaa cctcttacgt gccgatcaag tcaaaagcct ccggtcggag
     5701 gcttttgact ttctgctatg gaggtcaggt atgatttaaa tggtcagtta ga
//
