LOCUS       sequence-298064-        6014 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 6014)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6014)
  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-m4046217
FEATURES             Location/Qualifiers
     source          1..6014
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..418
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             complement(621..632)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             1689..1706
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1707..1724
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1769..1787)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2138..2941
                     /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      3024..4006
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4084..4113
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4135..4722
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4734..4761)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4785..4976)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4977..5768,5769..5837))
                     /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     5600..5619
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     gap             5870..5881
                     /estimated_length=12
     terminator      complement(5941..5972)
                     /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 nngaaaaagg caataaaaat ttttatcacg
      241 tttctttttc ttgaaaattt ttttttttng atttttttct ctttcgatga cctcccattg
      301 atatttaagt taataaacgg tcttcaattt ctcaagtttc agtttcattt ttcttgttct
      361 attacaactt tttttacttc ttgctcatta gaaagaaagc atagcaatct aatctaagtt
      421 ttaattacaa aatgtctgat aatggcccac agaaccaacg taacgcacct agaatcacct
      481 tcggtggtcc atccgacagc actggttcaa atcaaaacgg agagagatcg ggtgcccgga
      541 gtaagcaaag gcgcccccaa ggtctaccga ataacacagc ttcttggttt actgctctta
      601 cgcaacacgg taaagaagac ttgaagttcc caagaggtca aggggttcca attaacacca
      661 actcctctcc agatgatcaa ataggttact atagaagagc tactagacgt attaggggtg
      721 gggacggcaa aatgaaggat ttatctccta gatggtactt ttattacttg ggtaccggtc
      781 cagaagctgg attgccttac ggtgccaata aggacggtat catttgggta gcaactgaag
      841 gtgctttaaa taccccaaag gaccatatcg gtactagaaa ccctgctaat aacgctgcca
      901 ttgtcttgca attaccacag gggacaacgt tgccaaaggg tttctacgcc gaaggatcca
      961 gaggtggctc gcaagcttct agtagatcat cctctcgcag cagaaactca tctcgtaact
     1021 ccacaccagg tagttcgaga ggtacctctc cagcgagaat ggctggtaac ggtggtgatg
     1081 cagctctggc gttgctgcta ctcgacagat tgaatcaatt ggaatccaaa atgtctggta
     1141 agggtcaaca acaacagggt cagactgtga ctaagaagtc tgctgctgaa gcttccaaaa
     1201 agccccgtca aaagagaacc gccaccaagg cctataatgt cactcaagct ttcggaagaa
     1261 gaggtccaga gcaaacgcaa ggtaactttg gagatcaaga attgatccgg cagggtaccg
     1321 attataaaca ctggccccaa attgctcaat tcgcgccaag cgcctcagct tttttcggca
     1381 tgtcaagaat aggtatggaa gttactccat ctgggacttg gttaacttac actggtgcaa
     1441 tcaagttgga cgataaagac ccaaatttta aagaccaagt tattctactc aacaagcata
     1501 tagatgctta caaaaccttc ccgcctacag aacccaagaa agataaaaag aagaaggctg
     1561 acgagacaca agcattgcct cagcgtcaaa agaaacaaca aactgtcacc ttgttaccag
     1621 cagctgatct tgatgatttc tctaagcaat tgcaacaatc catgtcgtcc gccgactcta
     1681 ctcaagcgct cgtgccgaga ggttctcacc atcatcacca tcattaaaca ggcccctttt
     1741 cctttgtcga tatcatgtaa ttagttatgt cacgcttaca ttcacgccct cctcccacat
     1801 ccgctctaac cgaaaaggaa ggagttagac aacctgaagt ctaggtccct atttattttt
     1861 tttaatagtt atgttagtat taagaacgtt atttatattt caaatttttc ttttttttct
     1921 gtacaaagcg gccgccgaac cactagtgca ctgcagtaca cggtttcctt gaaatttttt
     1981 tgattcggta atctccgaac agaaggaaga acgaaggaag gagcacagac ttagattggt
     2041 atatatacgc atatgtagtg ttgaagaaac atgaaattgc ccagtattct taacccaact
     2101 gcacagaaca aaaacgtgca ggaaacgaag ataaatcatg tcgaaagcta catataagga
     2161 acgtgctgct actcatccta gtcctgttgc tgccaagcta tttaatatca tgcacgaaaa
     2221 gcaaacaaac ttgtgtgctt cattggatgt tcgtaccacc aaggaattac tggagttagt
     2281 tgaagcatta ggtcccaaaa tttgtttact aaaaacacat gtggatatct tgactgattt
     2341 ttccatggag ggcacagtta agccgctaaa ggcattatcc gccaagtaca attttttact
     2401 cttcgaggac agaaaatttg ctgacattgg taatacagtc aaattgcagt actctgcggg
     2461 tgtatacaga atagcagaat gggcagacat tacgaatgca cacggtgtgg tgggcccagg
     2521 tattgttagc ggtttgaagc aggcggcaga agaagtaaca aaggaaccta gaggcctttt
     2581 gatgttagca gaattgtcat gcaagggctc cctatctact ggagaatata ctaagggtac
     2641 tgttgacatt gcgaagagcg acaaagattt tgttatcggc tttattgctc aaagagacat
     2701 gggtggaaga gatgaaggtt acgattggtt gattatgaca cccggtgtgg gtttagatga
     2761 caagggagat gcattgggtc aacagtatag aaccgtggat gatgtggttt ctacaggatc
     2821 tgacattatt attgttggaa gaggactatt tgcaaaggga agggatgcta aggtagaggg
     2881 tgaacgttac agaaaagcag gctgggaagc atatttgaga agatgcggcc agcaaaacta
     2941 aaaaactgta ttataagtaa atgcatgtat actaaactca caaattagag cttcaattta
     3001 attatatcag ttattacccg agtaacgaag catctgtgct tcattttgta gaacaaaaat
     3061 gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga
     3121 aatgcaacgc gaaagcgcta ttttaccaac gaagaatctg tgcttcattt ttgtaaaaca
     3181 aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga
     3241 acagaaatgc aacgcgagag cgctatttta ccaacaaaga atctatactt cttttttgtt
     3301 ctacaaaaat gcatcccgag agcgctattt ttctaacaaa gcatcttaga ttactttttt
     3361 tctcctttgt gcgctctata atgcagtctc ttgataactt tttgcactgt aggtccgtta
     3421 aggttagaag aaggctactt tggtgtctat tttctcttcc ataaaaaaag cctgactcca
     3481 cttcccgcgt ttactgatta ctagcgaagc tgcgggtgca ttttttcaag ataaaggcat
     3541 ccccgattat attctatacc gatgtggatt gcgcatactt tgtgaacaga aagtgatagc
     3601 gttgatgatt cttcattggt cagaaaatta tgaacggttt cttctatttt gtctctatat
     3661 actacgtata ggaaatgttt acattttcgt attgttttcg attcactcta tgaatagttc
     3721 ttactacaat ttttttgtct aaagagtaat actagagata aacataaaaa atgtagaggt
     3781 cgagtttaga tgcaagttca aggagcgaaa ggtggatggg taggttatat agggatatag
     3841 cacagagata tatagcaaag agatactttt gagcaatgtt tgtggaagcg gtattcgcaa
     3901 tattttagta gctcgttaca gtccggtgcg tttttggttt tttgaaagtg cgtcatcaga
     3961 gcgcttttgg ttttcaaaag cgctctgaag ttcctatact ttctagctag agaataggaa
     4021 cttcccgagc ggccgcgtgt tacaagcttg actccagcgt aactggactg caatcaactc
     4081 actggctcac cttcacgggt gggcctttct tcggtagaaa atcaaaggat cttcttgaga
     4141 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     4201 ggtttgtttg ccggatcaag agctaccaac tctttttccg aggtaactgg cttcagcaga
     4261 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     4321 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     4381 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     4441 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     4501 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     4561 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     4621 ggggtaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcat
     4681 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcagaa
     4741 aggcccaccc gaaggtgagc caggtgatta catttgggcc ctcatcagag gttttcaccg
     4801 tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat
     4861 tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat
     4921 gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcacttac
     4981 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     5041 gcctgactcc ccgtcgtgta gataactacg atgcgggagg gcttaccatc tggccccagt
     5101 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     5161 ccagccggga gggccgagcg cagaagtgat cctgcaactt tatccgcctc catccagtct
     5221 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     5281 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     5341 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     5401 agctccttcg gtcctccgat cgttgccaga agtaagttgg ccgcagtgtt atcactcatg
     5461 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtgagatg cttttctgtg
     5521 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     5581 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     5641 attggaaaac gttcttcggg gcgtaaactc tcaaggatct taccgctgtt gagatccagt
     5701 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     5761 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     5821 aaatgttgaa tactcatttt agcttcctta gctcctgaaa atctcgatan nnnnnnnnnn
     5881 ncgcccggta gtgatcttat ttcattatgg tgaaagttgg aacctcttac gtgccgatca
     5941 agtcaaaagc ctccggtcgg aggcttttga ctttctgcta tggaggtcag gtatgattta
     6001 aatggtcagt taga
//
