LOCUS       sequence-291579-        6061 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 6061)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6061)
  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..6061
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             complement(620..631)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             1688..1705
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1706..1771
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(1816..1834)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2185..2988
                     /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      3071..4053
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4131..4160
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4182..4769
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4781..4808)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4832..5023)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5024..5815,5816..5884))
                     /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     5647..5666
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5885..5987)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5988..6019)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgccc cacacaccat agcttcaaaa tgtttctact ccttttttac tcttccagat
       61 tttctcggac tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag catactaaat
      121 ttcccctctt tcttcctcta gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa
      181 aaaagagacc gcctcgtttc tttttcttcg tcgaaaaagg caataaaaat ttttatcacg
      241 tttctttttc ttgaaaattt ttttttttga tttttttctc tttcgatgac ctcccattga
      301 tatttaagtt aataaacggt cttcaatttc tcaagtttca gtttcatttt tcttgttcta
      361 ttacaacttt ttttacttct tgctcattag aaagaaagca tagcaatcta atctaagttt
      421 taattacaaa atgtctgata atggcccaca gaaccaacgt aacgcaccta gaatcacctt
      481 cggtggtcca tccgacagca ctggttcaaa tcaaaacgga gagagatcgg gtgcccggag
      541 taagcaaagg cgcccccaag gtctaccgaa taacacagct tcttggttta ctgctcttac
      601 gcaacacggt aaagaagact tgaagttccc aagaggtcaa ggggttccaa ttaacaccaa
      661 ctcctctcca gatgatcaaa taggttacta tagaagagct actagacgta ttaggggtgg
      721 ggacggcaaa atgaaggatt tatctcctag atggtacttt tattacttgg gtaccggtcc
      781 agaagctgga ttgccttacg gtgccaataa ggacggtatc atttgggtag caactgaagg
      841 tgctttaaat accccaaagg accatatcgg tactagaaac cctgctaata acgctgccat
      901 tgtcttgcaa ttaccacagg ggacaacgtt gccaaagggt ttctacgccg aaggatccag
      961 aggtggctcg caagcttcta gtagatcatc ctctcgcagc agaaactcat ctcgtaactc
     1021 cacaccaggt agttcgagag gtacctctcc agcgagaatg gctggtaacg gtggtgatgc
     1081 agctctggcg ttgctgctac tcgacagatt gaatcaattg gaatccaaaa tgtctggtaa
     1141 gggtcaacaa caacagggtc agactgtgac taagaagtct gctgctgaag cttccaaaaa
     1201 gccccgtcaa aagagaaccg ccaccaaggc ctataatgtc actcaagctt tcggaagaag
     1261 aggtccagag caaacgcaag gtaactttgg agatcaagaa ttgatccggc agggtaccga
     1321 ttataaacac tggccccaaa ttgctcaatt cgcgccaagc gcctcagctt ttttcggcat
     1381 gtcaagaata ggtatggaag ttactccatc tgggacttgg ttaacttaca ctggtgcaat
     1441 caagttggac gataaagacc caaattttaa agaccaagtt attctactca acaagcatat
     1501 agatgcttac aaaaccttcc cgcctacaga acccaagaaa gataaaaaga agaaggctga
     1561 cgagacacaa gcattgcctc agcgtcaaaa gaaacaacaa actgtcacct tgttaccagc
     1621 agctgatctt gatgatttct ctaagcaatt gcaacaatcc atgtcgtccg ccgactctac
     1681 tcaagcgctc gtgccgagag gttctgatta taaagatcat gatggtgatt ataaagatca
     1741 tgatattgat tataaagatg atgatgataa ataaacaggc cccttttcct ttgtcgatat
     1801 catgtaatta gttatgtcac gcttacattc acgccctcct cccacatccg ctctaaccga
     1861 aaaggaagga gttagacaac ctgaagtcta ggtccctatt tatttttttt aatagttatg
     1921 ttagtattaa gaacgttatt tatatttcaa atttttcttt tttttctgta caaagcggcc
     1981 gccgaaccac tagtgcactg cagtacacgg tttccttgaa atttttttga ttcggtaatc
     2041 tccgaacaga aggaagaacg aaggaaggag cacagactta gattggtata tatacgcata
     2101 tgtagtgttg aagaaacatg aaattgccca gtattcttaa cccaactgca cagaacaaaa
     2161 acgtgcagga aacgaagata aatcatgtcg aaagctacat ataaggaacg tgctgctact
     2221 catcctagtc ctgttgctgc caagctattt aatatcatgc acgaaaagca aacaaacttg
     2281 tgtgcttcat tggatgttcg taccaccaag gaattactgg agttagttga agcattaggt
     2341 cccaaaattt gtttactaaa aacacatgtg gatatcttga ctgatttttc catggagggc
     2401 acagttaagc cgctaaaggc attatccgcc aagtacaatt ttttactctt cgaggacaga
     2461 aaatttgctg acattggtaa tacagtcaaa ttgcagtact ctgcgggtgt atacagaata
     2521 gcagaatggg cagacattac gaatgcacac ggtgtggtgg gcccaggtat tgttagcggt
     2581 ttgaagcagg cggcagaaga agtaacaaag gaacctagag gccttttgat gttagcagaa
     2641 ttgtcatgca agggctccct atctactgga gaatatacta agggtactgt tgacattgcg
     2701 aagagcgaca aagattttgt tatcggcttt attgctcaaa gagacatggg tggaagagat
     2761 gaaggttacg attggttgat tatgacaccc ggtgtgggtt tagatgacaa gggagatgca
     2821 ttgggtcaac agtatagaac cgtggatgat gtggtttcta caggatctga cattattatt
     2881 gttggaagag gactatttgc aaagggaagg gatgctaagg tagagggtga acgttacaga
     2941 aaagcaggct gggaagcata tttgagaaga tgcggccagc aaaactaaaa aactgtatta
     3001 taagtaaatg catgtatact aaactcacaa attagagctt caatttaatt atatcagtta
     3061 ttacccgagt aacgaagcat ctgtgcttca ttttgtagaa caaaaatgca acgcgagagc
     3121 gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgaa
     3181 agcgctattt taccaacgaa gaatctgtgc ttcatttttg taaaacaaaa atgcaacgcg
     3241 agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac
     3301 gcgagagcgc tattttacca acaaagaatc tatacttctt ttttgttcta caaaaatgca
     3361 tcccgagagc gctatttttc taacaaagca tcttagatta ctttttttct cctttgtgcg
     3421 ctctataatg cagtctcttg ataacttttt gcactgtagg tccgttaagg ttagaagaag
     3481 gctactttgg tgtctatttt ctcttccata aaaaaagcct gactccactt cccgcgttta
     3541 ctgattacta gcgaagctgc gggtgcattt tttcaagata aaggcatccc cgattatatt
     3601 ctataccgat gtggattgcg catactttgt gaacagaaag tgatagcgtt gatgattctt
     3661 cattggtcag aaaattatga acggtttctt ctattttgtc tctatatact acgtatagga
     3721 aatgtttaca ttttcgtatt gttttcgatt cactctatga atagttctta ctacaatttt
     3781 tttgtctaaa gagtaatact agagataaac ataaaaaatg tagaggtcga gtttagatgc
     3841 aagttcaagg agcgaaaggt ggatgggtag gttatatagg gatatagcac agagatatat
     3901 agcaaagaga tacttttgag caatgtttgt ggaagcggta ttcgcaatat tttagtagct
     3961 cgttacagtc cggtgcgttt ttggtttttt gaaagtgcgt catcagagcg cttttggttt
     4021 tcaaaagcgc tctgaagttc ctatactttc tagctagaga ataggaactt cccgagcggc
     4081 cgcgtgttac aagcttgact ccagcgtaac tggactgcaa tcaactcact ggctcacctt
     4141 cacgggtggg cctttcttcg gtagaaaatc aaaggatctt cttgagatcc tttttttctg
     4201 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4261 gatcaagagc taccaactct ttttccgagg taactggctt cagcagagcg cagataccaa
     4321 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     4381 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     4441 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     4501 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     4561 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     4621 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg gtaaacgcct
     4681 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcatcga tttttgtgat
     4741 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcagaaagg cccacccgaa
     4801 ggtgagccag gtgattacat ttgggccctc atcagaggtt ttcaccgtca tcaccgaaac
     4861 gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg
     4921 cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga
     4981 taaagcgggc catgttaagg gcggtttttt cctgtttggt cacttaccaa tgcttaatca
     5041 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     5101 tcgtgtagat aactacgatg cgggagggct taccatctgg ccccagtgct gcaatgatac
     5161 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccgggaggg
     5221 ccgagcgcag aagtgatcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     5281 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     5341 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5401 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5461 ctccgatcgt tgccagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5521 tgcataattc tcttactgtc atgccatccg tgagatgctt ttctgtgact ggtgagtact
     5581 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5641 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     5701 cttcggggcg taaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5761 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5821 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5881 tcattttagc ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg
     5941 atcttatttc attatggtga aagttggaac ctcttacgtg ccgatcaagt caaaagcctc
     6001 cggtcggagg cttttgactt tctgctatgg aggtcaggta tgatttaaat ggtcagttag
     6061 a
//
