LOCUS       sequence-291535-        7093 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 7093)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 7093)
  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-m4046575
FEATURES             Location/Qualifiers
     source          1..7093
                     /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"
     primer_bind     complement(2848..2866)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             3217..4020
                     /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      4103..5085
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      5163..5192
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      5214..5801
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5813..5840)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5864..6055)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6056..6847,6848..6916))
                     /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     6679..6698
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6917..7019)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(7020..7051)
                     /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 gttctaaaat cgaggaaggt aagctagtca tttggataaa
     1741 tggagataag ggttacaacg gtttggcaga agtaggcaag aaatttgaaa aggacacagg
     1801 tatcaaggtg acggttgaac atccagacaa gttagaagaa aaattccctc aagttgctgc
     1861 tactggtgat ggtcccgaca ttattttctg ggcccacgat cgttttggtg gatatgctca
     1921 gtcaggtttg ctggctgaga tcaccccaga caaggccttc caagataaat tgtacccatt
     1981 cacttgggat gctgtcagat acaacggtaa gcttatagca tatccaattg ctgttgaagc
     2041 cttatcgttg atctacaata aggacttgtt acctaaccca ccaaagacat gggaagaaat
     2101 tccagctttg gataaagagc taaaggccaa gggaaaaagc gctttgatgt ttaacttaca
     2161 agaaccttat ttcacctggc cattaatcgc tgctgacggt ggttacgcat ttaaatacga
     2221 aaacggtaag tatgacatta aggatgtcgg cgttgataat gctggtgcaa aggctggttt
     2281 gaccttcttg gttgatttaa taaagaataa acacatgaac gctgacactg actactctat
     2341 cgccgaagcc gctttcaaca agggtgaaac tgctatgacc ataaatggtc catgggcctg
     2401 gagtaacatt gatacttcta aagtaaacta tggtgtcact gttttgccta cattcaaagg
     2461 ccaaccatcc aagccctttg tcggtgttct atccgctggc atcaatgctg cttctccaaa
     2521 caaggagcta gctaaagaat ttttggaaaa ttacttgtta actgacgaag gtttggaggc
     2581 cgttaacaag gacaagccat tgggtgctgt cgctttgaaa tcttacgaag aagaattggc
     2641 taaggatccc agaattgccg ctaccatgga gaacgcccaa aagggtgaaa ttatgccaaa
     2701 catcccacaa atgtcagcct tctggtacgc agttaggact gccgtgatta atgctgcttc
     2761 cggtagacaa accgtcgacg aagcattaaa agatgctcaa acgtaaacag gccccttttc
     2821 ctttgtcgat atcatgtaat tagttatgtc acgcttacat tcacgccctc ctcccacatc
     2881 cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt
     2941 ttaatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg
     3001 tacaaagcgg ccgccgaacc actagtgcac tgcagtacac ggtttccttg aaattttttt
     3061 gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg agcacagact tagattggta
     3121 tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt aacccaactg
     3181 cacagaacaa aaacgtgcag gaaacgaaga taaatcatgt cgaaagctac atataaggaa
     3241 cgtgctgcta ctcatcctag tcctgttgct gccaagctat ttaatatcat gcacgaaaag
     3301 caaacaaact tgtgtgcttc attggatgtt cgtaccacca aggaattact ggagttagtt
     3361 gaagcattag gtcccaaaat ttgtttacta aaaacacatg tggatatctt gactgatttt
     3421 tccatggagg gcacagttaa gccgctaaag gcattatccg ccaagtacaa ttttttactc
     3481 ttcgaggaca gaaaatttgc tgacattggt aatacagtca aattgcagta ctctgcgggt
     3541 gtatacagaa tagcagaatg ggcagacatt acgaatgcac acggtgtggt gggcccaggt
     3601 attgttagcg gtttgaagca ggcggcagaa gaagtaacaa aggaacctag aggccttttg
     3661 atgttagcag aattgtcatg caagggctcc ctatctactg gagaatatac taagggtact
     3721 gttgacattg cgaagagcga caaagatttt gttatcggct ttattgctca aagagacatg
     3781 ggtggaagag atgaaggtta cgattggttg attatgacac ccggtgtggg tttagatgac
     3841 aagggagatg cattgggtca acagtataga accgtggatg atgtggtttc tacaggatct
     3901 gacattatta ttgttggaag aggactattt gcaaagggaa gggatgctaa ggtagagggt
     3961 gaacgttaca gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa
     4021 aaaactgtat tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa
     4081 ttatatcagt tattacccga gtaacgaagc atctgtgctt cattttgtag aacaaaaatg
     4141 caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa
     4201 atgcaacgcg aaagcgctat tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa
     4261 aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa
     4321 cagaaatgca acgcgagagc gctattttac caacaaagaa tctatacttc ttttttgttc
     4381 tacaaaaatg catcccgaga gcgctatttt tctaacaaag catcttagat tacttttttt
     4441 ctcctttgtg cgctctataa tgcagtctct tgataacttt ttgcactgta ggtccgttaa
     4501 ggttagaaga aggctacttt ggtgtctatt ttctcttcca taaaaaaagc ctgactccac
     4561 ttcccgcgtt tactgattac tagcgaagct gcgggtgcat tttttcaaga taaaggcatc
     4621 cccgattata ttctataccg atgtggattg cgcatacttt gtgaacagaa agtgatagcg
     4681 ttgatgattc ttcattggtc agaaaattat gaacggtttc ttctattttg tctctatata
     4741 ctacgtatag gaaatgttta cattttcgta ttgttttcga ttcactctat gaatagttct
     4801 tactacaatt tttttgtcta aagagtaata ctagagataa acataaaaaa tgtagaggtc
     4861 gagtttagat gcaagttcaa ggagcgaaag gtggatgggt aggttatata gggatatagc
     4921 acagagatat atagcaaaga gatacttttg agcaatgttt gtggaagcgg tattcgcaat
     4981 attttagtag ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc gtcatcagag
     5041 cgcttttggt tttcaaaagc gctctgaagt tcctatactt tctagctaga gaataggaac
     5101 ttcccgagcg gccgcgtgtt acaagcttga ctccagcgta actggactgc aatcaactca
     5161 ctggctcacc ttcacgggtg ggcctttctt cggtagaaaa tcaaaggatc ttcttgagat
     5221 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     5281 gtttgtttgc cggatcaaga gctaccaact ctttttccga ggtaactggc ttcagcagag
     5341 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     5401 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5461 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     5521 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     5581 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     5641 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     5701 gggtaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcatc
     5761 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcagaaa
     5821 ggcccacccg aaggtgagcc aggtgattac atttgggccc tcatcagagg ttttcaccgt
     5881 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt
     5941 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg
     6001 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcacttacc
     6061 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     6121 cctgactccc cgtcgtgtag ataactacga tgcgggaggg cttaccatct ggccccagtg
     6181 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     6241 cagccgggag ggccgagcgc agaagtgatc ctgcaacttt atccgcctcc atccagtcta
     6301 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     6361 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     6421 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     6481 gctccttcgg tcctccgatc gttgccagaa gtaagttggc cgcagtgtta tcactcatgg
     6541 ttatggcagc actgcataat tctcttactg tcatgccatc cgtgagatgc ttttctgtga
     6601 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     6661 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6721 ttggaaaacg ttcttcgggg cgtaaactct caaggatctt accgctgttg agatccagtt
     6781 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     6841 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     6901 aatgttgaat actcatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata
     6961 cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa
     7021 gtcaaaagcc tccggtcgga ggcttttgac tttctgctat ggaggtcagg tatgatttaa
     7081 atggtcagtt aga
//
