LOCUS       sequence-291583-        6066 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 6066)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6066)
  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-m4046548
FEATURES             Location/Qualifiers
     source          1..6066
                     /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             complement(667..678)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             1735..1752
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1753..1776
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1821..1839)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2190..2993
                     /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      3076..4058
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4136..4165
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4187..4774
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4786..4813)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4837..5028)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5029..5820,5821..5889))
                     /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     5652..5671
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5890..5992)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5993..6024)
                     /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 gcccacagaa ccaacgtaac gcacctagaa tcaccttcgg tggtccatcc
      541 gacagcactg gttcaaatca aaacggagag agatcgggtg cccggagtaa gcaaaggcgc
      601 ccccaaggtc taccgaataa cacagcttct tggtttactg ctcttacgca acacggtaaa
      661 gaagacttga agttcccaag aggtcaaggg gttccaatta acaccaactc ctctccagat
      721 gatcaaatag gttactatag aagagctact agacgtatta ggggtgggga cggcaaaatg
      781 aaggatttat ctcctagatg gtacttttat tacttgggta ccggtccaga agctggattg
      841 ccttacggtg ccaataagga cggtatcatt tgggtagcaa ctgaaggtgc tttaaatacc
      901 ccaaaggacc atatcggtac tagaaaccct gctaataacg ctgccattgt cttgcaatta
      961 ccacagggga caacgttgcc aaagggtttc tacgccgaag gatccagagg tggctcgcaa
     1021 gcttctagta gatcatcctc tcgcagcaga aactcatctc gtaactccac accaggtagt
     1081 tcgagaggta cctctccagc gagaatggct ggtaacggtg gtgatgcagc tctggcgttg
     1141 ctgctactcg acagattgaa tcaattggaa tccaaaatgt ctggtaaggg tcaacaacaa
     1201 cagggtcaga ctgtgactaa gaagtctgct gctgaagctt ccaaaaagcc ccgtcaaaag
     1261 agaaccgcca ccaaggccta taatgtcact caagctttcg gaagaagagg tccagagcaa
     1321 acgcaaggta actttggaga tcaagaattg atccggcagg gtaccgatta taaacactgg
     1381 ccccaaattg ctcaattcgc gccaagcgcc tcagcttttt tcggcatgtc aagaataggt
     1441 atggaagtta ctccatctgg gacttggtta acttacactg gtgcaatcaa gttggacgat
     1501 aaagacccaa attttaaaga ccaagttatt ctactcaaca agcatataga tgcttacaaa
     1561 accttcccgc ctacagaacc caagaaagat aaaaagaaga aggctgacga gacacaagca
     1621 ttgcctcagc gtcaaaagaa acaacaaact gtcaccttgt taccagcagc tgatcttgat
     1681 gatttctcta agcaattgca acaatccatg tcgtccgccg actctactca agcgctcgtg
     1741 ccgagaggtt cttggagcca tccgcagttt gaaaaataaa caggcccctt ttcctttgtc
     1801 gatatcatgt aattagttat gtcacgctta cattcacgcc ctcctcccac atccgctcta
     1861 accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt tttttaatag
     1921 ttatgttagt attaagaacg ttatttatat ttcaaatttt tctttttttt ctgtacaaag
     1981 cggccgccga accactagtg cactgcagta cacggtttcc ttgaaatttt tttgattcgg
     2041 taatctccga acagaaggaa gaacgaagga aggagcacag acttagattg gtatatatac
     2101 gcatatgtag tgttgaagaa acatgaaatt gcccagtatt cttaacccaa ctgcacagaa
     2161 caaaaacgtg caggaaacga agataaatca tgtcgaaagc tacatataag gaacgtgctg
     2221 ctactcatcc tagtcctgtt gctgccaagc tatttaatat catgcacgaa aagcaaacaa
     2281 acttgtgtgc ttcattggat gttcgtacca ccaaggaatt actggagtta gttgaagcat
     2341 taggtcccaa aatttgttta ctaaaaacac atgtggatat cttgactgat ttttccatgg
     2401 agggcacagt taagccgcta aaggcattat ccgccaagta caatttttta ctcttcgagg
     2461 acagaaaatt tgctgacatt ggtaatacag tcaaattgca gtactctgcg ggtgtataca
     2521 gaatagcaga atgggcagac attacgaatg cacacggtgt ggtgggccca ggtattgtta
     2581 gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc tagaggcctt ttgatgttag
     2641 cagaattgtc atgcaagggc tccctatcta ctggagaata tactaagggt actgttgaca
     2701 ttgcgaagag cgacaaagat tttgttatcg gctttattgc tcaaagagac atgggtggaa
     2761 gagatgaagg ttacgattgg ttgattatga cacccggtgt gggtttagat gacaagggag
     2821 atgcattggg tcaacagtat agaaccgtgg atgatgtggt ttctacagga tctgacatta
     2881 ttattgttgg aagaggacta tttgcaaagg gaagggatgc taaggtagag ggtgaacgtt
     2941 acagaaaagc aggctgggaa gcatatttga gaagatgcgg ccagcaaaac taaaaaactg
     3001 tattataagt aaatgcatgt atactaaact cacaaattag agcttcaatt taattatatc
     3061 agttattacc cgagtaacga agcatctgtg cttcattttg tagaacaaaa atgcaacgcg
     3121 agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac
     3181 gcgaaagcgc tattttacca acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca
     3241 acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat
     3301 gcaacgcgag agcgctattt taccaacaaa gaatctatac ttcttttttg ttctacaaaa
     3361 atgcatcccg agagcgctat ttttctaaca aagcatctta gattactttt tttctccttt
     3421 gtgcgctcta taatgcagtc tcttgataac tttttgcact gtaggtccgt taaggttaga
     3481 agaaggctac tttggtgtct attttctctt ccataaaaaa agcctgactc cacttcccgc
     3541 gtttactgat tactagcgaa gctgcgggtg cattttttca agataaaggc atccccgatt
     3601 atattctata ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata gcgttgatga
     3661 ttcttcattg gtcagaaaat tatgaacggt ttcttctatt ttgtctctat atactacgta
     3721 taggaaatgt ttacattttc gtattgtttt cgattcactc tatgaatagt tcttactaca
     3781 atttttttgt ctaaagagta atactagaga taaacataaa aaatgtagag gtcgagttta
     3841 gatgcaagtt caaggagcga aaggtggatg ggtaggttat atagggatat agcacagaga
     3901 tatatagcaa agagatactt ttgagcaatg tttgtggaag cggtattcgc aatattttag
     3961 tagctcgtta cagtccggtg cgtttttggt tttttgaaag tgcgtcatca gagcgctttt
     4021 ggttttcaaa agcgctctga agttcctata ctttctagct agagaatagg aacttcccga
     4081 gcggccgcgt gttacaagct tgactccagc gtaactggac tgcaatcaac tcactggctc
     4141 accttcacgg gtgggccttt cttcggtaga aaatcaaagg atcttcttga gatccttttt
     4201 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     4261 tgccggatca agagctacca actctttttc cgaggtaact ggcttcagca gagcgcagat
     4321 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4381 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4441 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4501 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4561 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4621 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc caggggtaaa
     4681 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc atcgattttt
     4741 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcag aaaggcccac
     4801 ccgaaggtga gccaggtgat tacatttggg ccctcatcag aggttttcac cgtcatcacc
     4861 gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat
     4921 gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct
     4981 tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactt accaatgctt
     5041 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     5101 ccccgtcgtg tagataacta cgatgcggga gggcttacca tctggcccca gtgctgcaat
     5161 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     5221 gagggccgag cgcagaagtg atcctgcaac tttatccgcc tccatccagt ctattaattg
     5281 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     5341 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     5401 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     5461 cggtcctccg atcgttgcca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     5521 agcactgcat aattctctta ctgtcatgcc atccgtgaga tgcttttctg tgactggtga
     5581 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     5641 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     5701 acgttcttcg gggcgtaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     5761 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     5821 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     5881 aatactcatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa atacgcccgg
     5941 tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat caagtcaaaa
     6001 gcctccggtc ggaggctttt gactttctgc tatggaggtc aggtatgatt taaatggtca
     6061 gttaga
//
