LOCUS       sequence-297571-        5977 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 5977)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5977)
  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-m4046696
FEATURES             Location/Qualifiers
     source          1..5977
                     /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             complement(620..631)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(1732..1750)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2101..2904
                     /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      2987..3969
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4047..4076
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4098..4685
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4697..4724)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4748..4939)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4940..5731,5732..5800))
                     /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     5563..5582
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5801..5903)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5904..5935)
                     /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 atggcccaca gaaccaacgt aacgcaccta gaatcacctt
      481 cggtggtcca tccgacagca ctggttcaaa tcaaaacgga gagagatcgg gtgcccggag
      541 taagcaaagg cgcccccaag gtctaccgaa taacacagct tcttggttta ctgctcttac
      601 gcaacacggt aaagaagact tgaagttccc aagaggtcaa ggggttccaa ttaacaccaa
      661 ctcctctcca gatgatcaaa taggttacta tagaagagct actagacgta ttaggggtgg
      721 ggacggcaaa atgaaggatt tatctcctag atggtacttt tattacttgg gtaccggtcc
      781 agaagctgga ttgccttacg gtgccaataa ggacggtatc atttgggtag caactgaagg
      841 tgctttaaat accccaaagg accatatcgg tactagaaac cctgctaata acgctgccat
      901 tgtcttgcaa ttaccacagg ggacaacgtt gccaaagggt ttctacgccg aaggatccag
      961 aggtggctcg caagcttcta gtagatcatc ctctcgcagc agaaactcat ctcgtaactc
     1021 cacaccaggt agttcgagag gtacctctcc agcgagaatg gctggtaacg gtggtgatgc
     1081 agctctggcg ttgctgctac tcgacagatt gaatcaattg gaatccaaaa tgtctggtaa
     1141 gggtcaacaa caacagggtc agactgtgac taagaagtct gctgctgaag cttccaaaaa
     1201 gccccgtcaa aagagaaccg ccaccaaggc ctataatgtc actcaagctt tcggaagaag
     1261 aggtccagag caaacgcaag gtaactttgg agatcaagaa ttgatccggc agggtaccga
     1321 ttataaacac tggccccaaa ttgctcaatt cgcgccaagc gcctcagctt ttttcggcat
     1381 gtcaagaata ggtatggaag ttactccatc tgggacttgg ttaacttaca ctggtgcaat
     1441 caagttggac gataaagacc caaattttaa agaccaagtt attctactca acaagcatat
     1501 agatgcttac aaaaccttcc cgcctacaga acccaagaaa gataaaaaga agaaggctga
     1561 cgagacacaa gcattgcctc agcgtcaaaa gaaacaacaa actgtcacct tgttaccagc
     1621 agctgatctt gatgatttct ctaagcaatt gcaacaatcc atgtcgtccg ccgactctac
     1681 tcaagcgtaa acaggcccct tttcctttgt cgatatcatg taattagtta tgtcacgctt
     1741 acattcacgc cctcctccca catccgctct aaccgaaaag gaaggagtta gacaacctga
     1801 agtctaggtc cctatttatt ttttttaata gttatgttag tattaagaac gttatttata
     1861 tttcaaattt ttcttttttt tctgtacaaa gcggccgccg aaccactagt gcactgcagt
     1921 acacggtttc cttgaaattt ttttgattcg gtaatctccg aacagaagga agaacgaagg
     1981 aaggagcaca gacttagatt ggtatatata cgcatatgta gtgttgaaga aacatgaaat
     2041 tgcccagtat tcttaaccca actgcacaga acaaaaacgt gcaggaaacg aagataaatc
     2101 atgtcgaaag ctacatataa ggaacgtgct gctactcatc ctagtcctgt tgctgccaag
     2161 ctatttaata tcatgcacga aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc
     2221 accaaggaat tactggagtt agttgaagca ttaggtccca aaatttgttt actaaaaaca
     2281 catgtggata tcttgactga tttttccatg gagggcacag ttaagccgct aaaggcatta
     2341 tccgccaagt acaatttttt actcttcgag gacagaaaat ttgctgacat tggtaataca
     2401 gtcaaattgc agtactctgc gggtgtatac agaatagcag aatgggcaga cattacgaat
     2461 gcacacggtg tggtgggccc aggtattgtt agcggtttga agcaggcggc agaagaagta
     2521 acaaaggaac ctagaggcct tttgatgtta gcagaattgt catgcaaggg ctccctatct
     2581 actggagaat atactaaggg tactgttgac attgcgaaga gcgacaaaga ttttgttatc
     2641 ggctttattg ctcaaagaga catgggtgga agagatgaag gttacgattg gttgattatg
     2701 acacccggtg tgggtttaga tgacaaggga gatgcattgg gtcaacagta tagaaccgtg
     2761 gatgatgtgg tttctacagg atctgacatt attattgttg gaagaggact atttgcaaag
     2821 ggaagggatg ctaaggtaga gggtgaacgt tacagaaaag caggctggga agcatatttg
     2881 agaagatgcg gccagcaaaa ctaaaaaact gtattataag taaatgcatg tatactaaac
     2941 tcacaaatta gagcttcaat ttaattatat cagttattac ccgagtaacg aagcatctgt
     3001 gcttcatttt gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct
     3061 gagctgcatt tttacagaac agaaatgcaa cgcgaaagcg ctattttacc aacgaagaat
     3121 ctgtgcttca tttttgtaaa acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag
     3181 aatctgagct gcatttttac agaacagaaa tgcaacgcga gagcgctatt ttaccaacaa
     3241 agaatctata cttctttttt gttctacaaa aatgcatccc gagagcgcta tttttctaac
     3301 aaagcatctt agattacttt ttttctcctt tgtgcgctct ataatgcagt ctcttgataa
     3361 ctttttgcac tgtaggtccg ttaaggttag aagaaggcta ctttggtgtc tattttctct
     3421 tccataaaaa aagcctgact ccacttcccg cgtttactga ttactagcga agctgcgggt
     3481 gcattttttc aagataaagg catccccgat tatattctat accgatgtgg attgcgcata
     3541 ctttgtgaac agaaagtgat agcgttgatg attcttcatt ggtcagaaaa ttatgaacgg
     3601 tttcttctat tttgtctcta tatactacgt ataggaaatg tttacatttt cgtattgttt
     3661 tcgattcact ctatgaatag ttcttactac aatttttttg tctaaagagt aatactagag
     3721 ataaacataa aaaatgtaga ggtcgagttt agatgcaagt tcaaggagcg aaaggtggat
     3781 gggtaggtta tatagggata tagcacagag atatatagca aagagatact tttgagcaat
     3841 gtttgtggaa gcggtattcg caatatttta gtagctcgtt acagtccggt gcgtttttgg
     3901 ttttttgaaa gtgcgtcatc agagcgcttt tggttttcaa aagcgctctg aagttcctat
     3961 actttctagc tagagaatag gaacttcccg agcggccgcg tgttacaagc ttgactccag
     4021 cgtaactgga ctgcaatcaa ctcactggct caccttcacg ggtgggcctt tcttcggtag
     4081 aaaatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     4141 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     4201 ccgaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     4261 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4321 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     4381 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     4441 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     4501 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     4561 gagagcgcac gagggagctt ccaggggtaa acgcctggta tctttatagt cctgtcgggt
     4621 ttcgccacct ctgacttgag catcgatttt tgtgatgctc gtcagggggg cggagcctat
     4681 ggaaaaacgc cagcaacgca gaaaggccca cccgaaggtg agccaggtga ttacatttgg
     4741 gccctcatca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct
     4801 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt
     4861 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg
     4921 ttttttcctg tttggtcact taccaatgct taatcagtga ggcacctatc tcagcgatct
     4981 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatgcggg
     5041 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     5101 cagatttatc agcaataaac cagccagccg ggagggccga gcgcagaagt gatcctgcaa
     5161 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     5221 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     5281 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     5341 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgcc agaagtaagt
     5401 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     5461 catccgtgag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     5521 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     5581 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgtaaa ctctcaagga
     5641 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     5701 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     5761 aaaagggaat aagggcgaca cggaaatgtt gaatactcat tttagcttcc ttagctcctg
     5821 aaaatctcga taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt
     5881 tggaacctct tacgtgccga tcaagtcaaa agcctccggt cggaggcttt tgactttctg
     5941 ctatggaggt caggtatgat ttaaatggtc agttaga
//
