LOCUS       sequence-297782-        7140 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 7140)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 7140)
  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-m4046336
FEATURES             Location/Qualifiers
     source          1..7140
                     /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"
     primer_bind     complement(2895..2913)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             3264..4067
                     /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      4150..5132
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      5210..5239
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      5261..5848
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5860..5887)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5911..6102)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6103..6894,6895..6963))
                     /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     6726..6745
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6964..7066)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(7067..7098)
                     /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 ctaaaatcga ggaaggtaag ctagtcattt ggataaatgg agataagggt
     1801 tacaacggtt tggcagaagt aggcaagaaa tttgaaaagg acacaggtat caaggtgacg
     1861 gttgaacatc cagacaagtt agaagaaaaa ttccctcaag ttgctgctac tggtgatggt
     1921 cccgacatta ttttctgggc ccacgatcgt tttggtggat atgctcagtc aggtttgctg
     1981 gctgagatca ccccagacaa ggccttccaa gataaattgt acccattcac ttgggatgct
     2041 gtcagataca acggtaagct tatagcatat ccaattgctg ttgaagcctt atcgttgatc
     2101 tacaataagg acttgttacc taacccacca aagacatggg aagaaattcc agctttggat
     2161 aaagagctaa aggccaaggg aaaaagcgct ttgatgttta acttacaaga accttatttc
     2221 acctggccat taatcgctgc tgacggtggt tacgcattta aatacgaaaa cggtaagtat
     2281 gacattaagg atgtcggcgt tgataatgct ggtgcaaagg ctggtttgac cttcttggtt
     2341 gatttaataa agaataaaca catgaacgct gacactgact actctatcgc cgaagccgct
     2401 ttcaacaagg gtgaaactgc tatgaccata aatggtccat gggcctggag taacattgat
     2461 acttctaaag taaactatgg tgtcactgtt ttgcctacat tcaaaggcca accatccaag
     2521 ccctttgtcg gtgttctatc cgctggcatc aatgctgctt ctccaaacaa ggagctagct
     2581 aaagaatttt tggaaaatta cttgttaact gacgaaggtt tggaggccgt taacaaggac
     2641 aagccattgg gtgctgtcgc tttgaaatct tacgaagaag aattggctaa ggatcccaga
     2701 attgccgcta ccatggagaa cgcccaaaag ggtgaaatta tgccaaacat cccacaaatg
     2761 tcagccttct ggtacgcagt taggactgcc gtgattaatg ctgcttccgg tagacaaacc
     2821 gtcgacgaag cattaaaaga tgctcaaacg taaacaggcc ccttttcctt tgtcgatatc
     2881 atgtaattag ttatgtcacg cttacattca cgccctcctc ccacatccgc tctaaccgaa
     2941 aaggaaggag ttagacaacc tgaagtctag gtccctattt atttttttta atagttatgt
     3001 tagtattaag aacgttattt atatttcaaa tttttctttt ttttctgtac aaagcggccg
     3061 ccgaaccact agtgcactgc agtacacggt ttccttgaaa tttttttgat tcggtaatct
     3121 ccgaacagaa ggaagaacga aggaaggagc acagacttag attggtatat atacgcatat
     3181 gtagtgttga agaaacatga aattgcccag tattcttaac ccaactgcac agaacaaaaa
     3241 cgtgcaggaa acgaagataa atcatgtcga aagctacata taaggaacgt gctgctactc
     3301 atcctagtcc tgttgctgcc aagctattta atatcatgca cgaaaagcaa acaaacttgt
     3361 gtgcttcatt ggatgttcgt accaccaagg aattactgga gttagttgaa gcattaggtc
     3421 ccaaaatttg tttactaaaa acacatgtgg atatcttgac tgatttttcc atggagggca
     3481 cagttaagcc gctaaaggca ttatccgcca agtacaattt tttactcttc gaggacagaa
     3541 aatttgctga cattggtaat acagtcaaat tgcagtactc tgcgggtgta tacagaatag
     3601 cagaatgggc agacattacg aatgcacacg gtgtggtggg cccaggtatt gttagcggtt
     3661 tgaagcaggc ggcagaagaa gtaacaaagg aacctagagg ccttttgatg ttagcagaat
     3721 tgtcatgcaa gggctcccta tctactggag aatatactaa gggtactgtt gacattgcga
     3781 agagcgacaa agattttgtt atcggcttta ttgctcaaag agacatgggt ggaagagatg
     3841 aaggttacga ttggttgatt atgacacccg gtgtgggttt agatgacaag ggagatgcat
     3901 tgggtcaaca gtatagaacc gtggatgatg tggtttctac aggatctgac attattattg
     3961 ttggaagagg actatttgca aagggaaggg atgctaaggt agagggtgaa cgttacagaa
     4021 aagcaggctg ggaagcatat ttgagaagat gcggccagca aaactaaaaa actgtattat
     4081 aagtaaatgc atgtatacta aactcacaaa ttagagcttc aatttaatta tatcagttat
     4141 tacccgagta acgaagcatc tgtgcttcat tttgtagaac aaaaatgcaa cgcgagagcg
     4201 ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg caacgcgaaa
     4261 gcgctatttt accaacgaag aatctgtgct tcatttttgt aaaacaaaaa tgcaacgcga
     4321 gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg
     4381 cgagagcgct attttaccaa caaagaatct atacttcttt tttgttctac aaaaatgcat
     4441 cccgagagcg ctatttttct aacaaagcat cttagattac tttttttctc ctttgtgcgc
     4501 tctataatgc agtctcttga taactttttg cactgtaggt ccgttaaggt tagaagaagg
     4561 ctactttggt gtctattttc tcttccataa aaaaagcctg actccacttc ccgcgtttac
     4621 tgattactag cgaagctgcg ggtgcatttt ttcaagataa aggcatcccc gattatattc
     4681 tataccgatg tggattgcgc atactttgtg aacagaaagt gatagcgttg atgattcttc
     4741 attggtcaga aaattatgaa cggtttcttc tattttgtct ctatatacta cgtataggaa
     4801 atgtttacat tttcgtattg ttttcgattc actctatgaa tagttcttac tacaattttt
     4861 ttgtctaaag agtaatacta gagataaaca taaaaaatgt agaggtcgag tttagatgca
     4921 agttcaagga gcgaaaggtg gatgggtagg ttatataggg atatagcaca gagatatata
     4981 gcaaagagat acttttgagc aatgtttgtg gaagcggtat tcgcaatatt ttagtagctc
     5041 gttacagtcc ggtgcgtttt tggttttttg aaagtgcgtc atcagagcgc ttttggtttt
     5101 caaaagcgct ctgaagttcc tatactttct agctagagaa taggaacttc ccgagcggcc
     5161 gcgtgttaca agcttgactc cagcgtaact ggactgcaat caactcactg gctcaccttc
     5221 acgggtgggc ctttcttcgg tagaaaatca aaggatcttc ttgagatcct ttttttctgc
     5281 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     5341 atcaagagct accaactctt tttccgaggt aactggcttc agcagagcgc agataccaaa
     5401 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     5461 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     5521 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     5581 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     5641 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     5701 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg taaacgcctg
     5761 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcatcgat ttttgtgatg
     5821 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcagaaaggc ccacccgaag
     5881 gtgagccagg tgattacatt tgggccctca tcagaggttt tcaccgtcat caccgaaacg
     5941 cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc
     6001 ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat
     6061 aaagcgggcc atgttaaggg cggttttttc ctgtttggtc acttaccaat gcttaatcag
     6121 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6181 cgtgtagata actacgatgc gggagggctt accatctggc cccagtgctg caatgatacc
     6241 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccgggagggc
     6301 cgagcgcaga agtgatcctg caactttatc cgcctccatc cagtctatta attgttgccg
     6361 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     6421 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     6481 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6541 tccgatcgtt gccagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     6601 gcataattct cttactgtca tgccatccgt gagatgcttt tctgtgactg gtgagtactc
     6661 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6721 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6781 ttcggggcgt aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6841 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6901 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6961 cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga
     7021 tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaagtc aaaagcctcc
     7081 ggtcggaggc ttttgacttt ctgctatgga ggtcaggtat gatttaaatg gtcagttaga
//
