LOCUS       sequence-300250-        6060 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 6060)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6060)
  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-m4046295
FEATURES             Location/Qualifiers
     source          1..6060
                     /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             1735..1752
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1753..1770
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1815..1833)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2184..2987
                     /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      3070..4052
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4130..4159
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4181..4768
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4780..4807)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4831..5022)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5023..5814,5815..5883))
                     /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     5646..5665
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5884..5986)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5987..6018)
                     /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 gaccccaaaa tcagcgaaat gcaccccgca ttacgtttgg tggaccctca
      541 gattcaactg gcagtaacca gaatggagaa cgcagtgggg cgcgatcaaa acaacgtcgg
      601 ccccaaggtt tacccaataa tactgcgtct tggttcaccg ctctcactca acatggcaag
      661 gaagacctta aattccctcg aggacaaggc gttccaatta acaccaatag cagtccagat
      721 gaccaaattg gctactaccg aagagctacc agacgaattc gtggtggtga cggtaaaatg
      781 aaagatctca gtccaagatg gtatttctac tacctaggaa ctgggccaga agctggactt
      841 ccctatggtg ctaacaaaga cggcatcata tgggttgcaa ctgagggagc cttgaataca
      901 ccaaaagatc acattggcac ccgcaatcct gctaacaatg ctgcaatcgt gctacaactt
      961 cctcaaggaa caacattgcc aaaaggcttc tacgcagaag ggagcagagg cggcagtcaa
     1021 gcctcttctc gttcctcatc acgtagtcgc aacagttcaa gaaattcaac tccaggcagc
     1081 agtaggggaa cttctcctgc tagaatggct ggcaatggcg gtgatgctgc tcttgctttg
     1141 ctgctgcttg acagattgaa ccagcttgag agcaaaatgt ctggtaaagg ccaacaacaa
     1201 caaggccaaa ctgtcactaa gaaatctgct gctgaggctt ctaagaagcc tcggcaaaaa
     1261 cgtactgcca ctaaagcata caatgtaaca caagctttcg gcagacgtgg tccagaacaa
     1321 acccaaggaa attttgggga ccaggaacta atcagacaag gaactgatta caaacattgg
     1381 ccgcaaattg cacaatttgc ccccagcgct tcagcgttct tcggaatgtc gcgcattggc
     1441 atggaagtca caccttcggg aacgtggttg acctacacag gtgccatcaa attggatgac
     1501 aaagatccaa atttcaaaga tcaagtcatt ttgctgaata agcatattga cgcatacaaa
     1561 acattcccac caacagagcc taaaaaggac aaaaagaaga aggctgatga aactcaagcc
     1621 ttaccgcaga gacagaagaa acagcaaact gtgactcttc ttcctgctgc agatttggat
     1681 gatttctcca aacaattgca acaatccatg agcagtgctg actcaactca ggccctcgtg
     1741 ccgagaggtt ctcaccatca tcaccatcat taaacaggcc ccttttcctt tgtcgatatc
     1801 atgtaattag ttatgtcacg cttacattca cgccctcctc ccacatccgc tctaaccgaa
     1861 aaggaaggag ttagacaacc tgaagtctag gtccctattt atttttttta atagttatgt
     1921 tagtattaag aacgttattt atatttcaaa tttttctttt ttttctgtac aaagcggccg
     1981 ccgaaccact agtgcactgc agtacacggt ttccttgaaa tttttttgat tcggtaatct
     2041 ccgaacagaa ggaagaacga aggaaggagc acagacttag attggtatat atacgcatat
     2101 gtagtgttga agaaacatga aattgcccag tattcttaac ccaactgcac agaacaaaaa
     2161 cgtgcaggaa acgaagataa atcatgtcga aagctacata taaggaacgt gctgctactc
     2221 atcctagtcc tgttgctgcc aagctattta atatcatgca cgaaaagcaa acaaacttgt
     2281 gtgcttcatt ggatgttcgt accaccaagg aattactgga gttagttgaa gcattaggtc
     2341 ccaaaatttg tttactaaaa acacatgtgg atatcttgac tgatttttcc atggagggca
     2401 cagttaagcc gctaaaggca ttatccgcca agtacaattt tttactcttc gaggacagaa
     2461 aatttgctga cattggtaat acagtcaaat tgcagtactc tgcgggtgta tacagaatag
     2521 cagaatgggc agacattacg aatgcacacg gtgtggtggg cccaggtatt gttagcggtt
     2581 tgaagcaggc ggcagaagaa gtaacaaagg aacctagagg ccttttgatg ttagcagaat
     2641 tgtcatgcaa gggctcccta tctactggag aatatactaa gggtactgtt gacattgcga
     2701 agagcgacaa agattttgtt atcggcttta ttgctcaaag agacatgggt ggaagagatg
     2761 aaggttacga ttggttgatt atgacacccg gtgtgggttt agatgacaag ggagatgcat
     2821 tgggtcaaca gtatagaacc gtggatgatg tggtttctac aggatctgac attattattg
     2881 ttggaagagg actatttgca aagggaaggg atgctaaggt agagggtgaa cgttacagaa
     2941 aagcaggctg ggaagcatat ttgagaagat gcggccagca aaactaaaaa actgtattat
     3001 aagtaaatgc atgtatacta aactcacaaa ttagagcttc aatttaatta tatcagttat
     3061 tacccgagta acgaagcatc tgtgcttcat tttgtagaac aaaaatgcaa cgcgagagcg
     3121 ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg caacgcgaaa
     3181 gcgctatttt accaacgaag aatctgtgct tcatttttgt aaaacaaaaa tgcaacgcga
     3241 gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg
     3301 cgagagcgct attttaccaa caaagaatct atacttcttt tttgttctac aaaaatgcat
     3361 cccgagagcg ctatttttct aacaaagcat cttagattac tttttttctc ctttgtgcgc
     3421 tctataatgc agtctcttga taactttttg cactgtaggt ccgttaaggt tagaagaagg
     3481 ctactttggt gtctattttc tcttccataa aaaaagcctg actccacttc ccgcgtttac
     3541 tgattactag cgaagctgcg ggtgcatttt ttcaagataa aggcatcccc gattatattc
     3601 tataccgatg tggattgcgc atactttgtg aacagaaagt gatagcgttg atgattcttc
     3661 attggtcaga aaattatgaa cggtttcttc tattttgtct ctatatacta cgtataggaa
     3721 atgtttacat tttcgtattg ttttcgattc actctatgaa tagttcttac tacaattttt
     3781 ttgtctaaag agtaatacta gagataaaca taaaaaatgt agaggtcgag tttagatgca
     3841 agttcaagga gcgaaaggtg gatgggtagg ttatataggg atatagcaca gagatatata
     3901 gcaaagagat acttttgagc aatgtttgtg gaagcggtat tcgcaatatt ttagtagctc
     3961 gttacagtcc ggtgcgtttt tggttttttg aaagtgcgtc atcagagcgc ttttggtttt
     4021 caaaagcgct ctgaagttcc tatactttct agctagagaa taggaacttc ccgagcggcc
     4081 gcgtgttaca agcttgactc cagcgtaact ggactgcaat caactcactg gctcaccttc
     4141 acgggtgggc ctttcttcgg tagaaaatca aaggatcttc ttgagatcct ttttttctgc
     4201 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     4261 atcaagagct accaactctt tttccgaggt aactggcttc agcagagcgc agataccaaa
     4321 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4381 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4441 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4501 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     4561 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4621 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg taaacgcctg
     4681 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcatcgat ttttgtgatg
     4741 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcagaaaggc ccacccgaag
     4801 gtgagccagg tgattacatt tgggccctca tcagaggttt tcaccgtcat caccgaaacg
     4861 cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc
     4921 ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat
     4981 aaagcgggcc atgttaaggg cggttttttc ctgtttggtc acttaccaat gcttaatcag
     5041 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5101 cgtgtagata actacgatgc gggagggctt accatctggc cccagtgctg caatgatacc
     5161 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccgggagggc
     5221 cgagcgcaga agtgatcctg caactttatc cgcctccatc cagtctatta attgttgccg
     5281 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     5341 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     5401 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     5461 tccgatcgtt gccagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     5521 gcataattct cttactgtca tgccatccgt gagatgcttt tctgtgactg gtgagtactc
     5581 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     5641 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     5701 ttcggggcgt aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     5761 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     5821 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     5881 cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga
     5941 tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaagtc aaaagcctcc
     6001 ggtcggaggc ttttgacttt ctgctatgga ggtcaggtat gatttaaatg gtcagttaga
//
