LOCUS       sequence-292474-        6019 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 6019)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6019)
  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-m4046588
FEATURES             Location/Qualifiers
     source          1..6019
                     /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             1688..1705
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1706..1729
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1774..1792)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2143..2946
                     /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      3029..4011
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4089..4118
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4140..4727
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4739..4766)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4790..4981)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4982..5773,5774..5842))
                     /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     5605..5624
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5843..5945)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5946..5977)
                     /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 atggacccca aaatcagcga aatgcacccc gcattacgtt
      481 tggtggaccc tcagattcaa ctggcagtaa ccagaatgga gaacgcagtg gggcgcgatc
      541 aaaacaacgt cggccccaag gtttacccaa taatactgcg tcttggttca ccgctctcac
      601 tcaacatggc aaggaagacc ttaaattccc tcgaggacaa ggcgttccaa ttaacaccaa
      661 tagcagtcca gatgaccaaa ttggctacta ccgaagagct accagacgaa ttcgtggtgg
      721 tgacggtaaa atgaaagatc tcagtccaag atggtatttc tactacctag gaactgggcc
      781 agaagctgga cttccctatg gtgctaacaa agacggcatc atatgggttg caactgaggg
      841 agccttgaat acaccaaaag atcacattgg cacccgcaat cctgctaaca atgctgcaat
      901 cgtgctacaa cttcctcaag gaacaacatt gccaaaaggc ttctacgcag aagggagcag
      961 aggcggcagt caagcctctt ctcgttcctc atcacgtagt cgcaacagtt caagaaattc
     1021 aactccaggc agcagtaggg gaacttctcc tgctagaatg gctggcaatg gcggtgatgc
     1081 tgctcttgct ttgctgctgc ttgacagatt gaaccagctt gagagcaaaa tgtctggtaa
     1141 aggccaacaa caacaaggcc aaactgtcac taagaaatct gctgctgagg cttctaagaa
     1201 gcctcggcaa aaacgtactg ccactaaagc atacaatgta acacaagctt tcggcagacg
     1261 tggtccagaa caaacccaag gaaattttgg ggaccaggaa ctaatcagac aaggaactga
     1321 ttacaaacat tggccgcaaa ttgcacaatt tgcccccagc gcttcagcgt tcttcggaat
     1381 gtcgcgcatt ggcatggaag tcacaccttc gggaacgtgg ttgacctaca caggtgccat
     1441 caaattggat gacaaagatc caaatttcaa agatcaagtc attttgctga ataagcatat
     1501 tgacgcatac aaaacattcc caccaacaga gcctaaaaag gacaaaaaga agaaggctga
     1561 tgaaactcaa gccttaccgc agagacagaa gaaacagcaa actgtgactc ttcttcctgc
     1621 tgcagatttg gatgatttct ccaaacaatt gcaacaatcc atgagcagtg ctgactcaac
     1681 tcaggccctc gtgccgagag gttcttggag ccatccgcag tttgaaaaat aaacaggccc
     1741 cttttccttt gtcgatatca tgtaattagt tatgtcacgc ttacattcac gccctcctcc
     1801 cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta
     1861 ttttttttaa tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt
     1921 tttctgtaca aagcggccgc cgaaccacta gtgcactgca gtacacggtt tccttgaaat
     1981 ttttttgatt cggtaatctc cgaacagaag gaagaacgaa ggaaggagca cagacttaga
     2041 ttggtatata tacgcatatg tagtgttgaa gaaacatgaa attgcccagt attcttaacc
     2101 caactgcaca gaacaaaaac gtgcaggaaa cgaagataaa tcatgtcgaa agctacatat
     2161 aaggaacgtg ctgctactca tcctagtcct gttgctgcca agctatttaa tatcatgcac
     2221 gaaaagcaaa caaacttgtg tgcttcattg gatgttcgta ccaccaagga attactggag
     2281 ttagttgaag cattaggtcc caaaatttgt ttactaaaaa cacatgtgga tatcttgact
     2341 gatttttcca tggagggcac agttaagccg ctaaaggcat tatccgccaa gtacaatttt
     2401 ttactcttcg aggacagaaa atttgctgac attggtaata cagtcaaatt gcagtactct
     2461 gcgggtgtat acagaatagc agaatgggca gacattacga atgcacacgg tgtggtgggc
     2521 ccaggtattg ttagcggttt gaagcaggcg gcagaagaag taacaaagga acctagaggc
     2581 cttttgatgt tagcagaatt gtcatgcaag ggctccctat ctactggaga atatactaag
     2641 ggtactgttg acattgcgaa gagcgacaaa gattttgtta tcggctttat tgctcaaaga
     2701 gacatgggtg gaagagatga aggttacgat tggttgatta tgacacccgg tgtgggttta
     2761 gatgacaagg gagatgcatt gggtcaacag tatagaaccg tggatgatgt ggtttctaca
     2821 ggatctgaca ttattattgt tggaagagga ctatttgcaa agggaaggga tgctaaggta
     2881 gagggtgaac gttacagaaa agcaggctgg gaagcatatt tgagaagatg cggccagcaa
     2941 aactaaaaaa ctgtattata agtaaatgca tgtatactaa actcacaaat tagagcttca
     3001 atttaattat atcagttatt acccgagtaa cgaagcatct gtgcttcatt ttgtagaaca
     3061 aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga
     3121 acagaaatgc aacgcgaaag cgctatttta ccaacgaaga atctgtgctt catttttgta
     3181 aaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt
     3241 acagaacaga aatgcaacgc gagagcgcta ttttaccaac aaagaatcta tacttctttt
     3301 ttgttctaca aaaatgcatc ccgagagcgc tatttttcta acaaagcatc ttagattact
     3361 ttttttctcc tttgtgcgct ctataatgca gtctcttgat aactttttgc actgtaggtc
     3421 cgttaaggtt agaagaaggc tactttggtg tctattttct cttccataaa aaaagcctga
     3481 ctccacttcc cgcgtttact gattactagc gaagctgcgg gtgcattttt tcaagataaa
     3541 ggcatccccg attatattct ataccgatgt ggattgcgca tactttgtga acagaaagtg
     3601 atagcgttga tgattcttca ttggtcagaa aattatgaac ggtttcttct attttgtctc
     3661 tatatactac gtataggaaa tgtttacatt ttcgtattgt tttcgattca ctctatgaat
     3721 agttcttact acaatttttt tgtctaaaga gtaatactag agataaacat aaaaaatgta
     3781 gaggtcgagt ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt tatataggga
     3841 tatagcacag agatatatag caaagagata cttttgagca atgtttgtgg aagcggtatt
     3901 cgcaatattt tagtagctcg ttacagtccg gtgcgttttt ggttttttga aagtgcgtca
     3961 tcagagcgct tttggttttc aaaagcgctc tgaagttcct atactttcta gctagagaat
     4021 aggaacttcc cgagcggccg cgtgttacaa gcttgactcc agcgtaactg gactgcaatc
     4081 aactcactgg ctcaccttca cgggtgggcc tttcttcggt agaaaatcaa aggatcttct
     4141 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     4201 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaggta actggcttca
     4261 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     4321 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     4381 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     4441 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     4501 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     4561 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     4621 ttccaggggt aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     4681 agcatcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     4741 cagaaaggcc cacccgaagg tgagccaggt gattacattt gggccctcat cagaggtttt
     4801 caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa
     4861 gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg
     4921 ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca
     4981 cttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5041 tagttgcctg actccccgtc gtgtagataa ctacgatgcg ggagggctta ccatctggcc
     5101 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5161 accagccagc cgggagggcc gagcgcagaa gtgatcctgc aactttatcc gcctccatcc
     5221 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5281 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5341 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5401 cggttagctc cttcggtcct ccgatcgttg ccagaagtaa gttggccgca gtgttatcac
     5461 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgtg agatgctttt
     5521 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5581 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5641 tcatcattgg aaaacgttct tcggggcgta aactctcaag gatcttaccg ctgttgagat
     5701 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     5761 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     5821 cacggaaatg ttgaatactc attttagctt ccttagctcc tgaaaatctc gataactcaa
     5881 aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc
     5941 gatcaagtca aaagcctccg gtcggaggct tttgactttc tgctatggag gtcaggtatg
     6001 atttaaatgg tcagttaga
//
