LOCUS       sequence-296253-        6024 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 6024)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6024)
  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-m4046661
FEATURES             Location/Qualifiers
     source          1..6024
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        50..491
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    50..167
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     462..485
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     complement(1804..1822)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2173..2976
                     /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      3059..4041
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4119..4148
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4170..4757
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4769..4796)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4820..5011)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5012..5803,5804..5872))
                     /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     5635..5654
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5873..5975)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5976..6007)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 ggtatgattt aaatggtcag ttagagcggc cgcttgctat attgaagtac ggattagaag
       61 ccgccgagcg ggcgacagcc ctccgacgga agactctcct ccgtgcgtcc tcgtcttcac
      121 cggtcgcgtt cctgaaacgc agatgtgcct cgcgccgcac tgctccgaac aataaagatt
      181 ctacaatact agcttttatg gttatgaaga ggaaaaattg gcagtaacct ggccccacaa
      241 accttcaaat taacgaatca aattaacaac cataggatga taatgcgatt agttttttag
      301 ccttatttct ggggtaatta atcagcgaag cgatgatttt tgatctatta acagatatat
      361 aaatggaaaa gctgcataac cactttaact aatactttca acattttcag tttgtattac
      421 ttcttattca aatgtcataa aagtatcaac aaaaaattgt taatatacct ctatacttta
      481 acgtcaagga gaaaaaacta taatgtctga taatggaccc caaaatcagc gaaatgcacc
      541 ccgcattacg tttggtggac cctcagattc aactggcagt aaccagaatg gagaacgcag
      601 tggggcgcga tcaaaacaac gtcggcccca aggtttaccc aataatactg cgtcttggtt
      661 caccgctctc actcaacatg gcaaggaaga ccttaaattc cctcgaggac aaggcgttcc
      721 aattaacacc aatagcagtc cagatgacca aattggctac taccgaagag ctaccagacg
      781 aattcgtggt ggtgacggta aaatgaaaga tctcagtcca agatggtatt tctactacct
      841 aggaactggg ccagaagctg gacttcccta tggtgctaac aaagacggca tcatatgggt
      901 tgcaactgag ggagccttga atacaccaaa agatcacatt ggcacccgca atcctgctaa
      961 caatgctgca atcgtgctac aacttcctca aggaacaaca ttgccaaaag gcttctacgc
     1021 agaagggagc agaggcggca gtcaagcctc ttctcgttcc tcatcacgta gtcgcaacag
     1081 ttcaagaaat tcaactccag gcagcagtag gggaacttct cctgctagaa tggctggcaa
     1141 tggcggtgat gctgctcttg ctttgctgct gcttgacaga ttgaaccagc ttgagagcaa
     1201 aatgtctggt aaaggccaac aacaacaagg ccaaactgtc actaagaaat ctgctgctga
     1261 ggcttctaag aagcctcggc aaaaacgtac tgccactaaa gcatacaatg taacacaagc
     1321 tttcggcaga cgtggtccag aacaaaccca aggaaatttt ggggaccagg aactaatcag
     1381 acaaggaact gattacaaac attggccgca aattgcacaa tttgccccca gcgcttcagc
     1441 gttcttcgga atgtcgcgca ttggcatgga agtcacacct tcgggaacgt ggttgaccta
     1501 cacaggtgcc atcaaattgg atgacaaaga tccaaatttc aaagatcaag tcattttgct
     1561 gaataagcat attgacgcat acaaaacatt cccaccaaca gagcctaaaa aggacaaaaa
     1621 gaagaaggct gatgaaactc aagccttacc gcagagacag aagaaacagc aaactgtgac
     1681 tcttcttcct gctgcagatt tggatgattt ctccaaacaa ttgcaacaat ccatgagcag
     1741 tgctgactca actcaggcct aaacaggccc cttttccttt gtcgatatca tgtaattagt
     1801 tatgtcacgc ttacattcac gccctcctcc cacatccgct ctaaccgaaa aggaaggagt
     1861 tagacaacct gaagtctagg tccctattta ttttttttaa tagttatgtt agtattaaga
     1921 acgttattta tatttcaaat ttttcttttt tttctgtaca aagcggccgc cgaaccacta
     1981 gtgcactgca gtacacggtt tccttgaaat ttttttgatt cggtaatctc cgaacagaag
     2041 gaagaacgaa ggaaggagca cagacttaga ttggtatata tacgcatatg tagtgttgaa
     2101 gaaacatgaa attgcccagt attcttaacc caactgcaca gaacaaaaac gtgcaggaaa
     2161 cgaagataaa tcatgtcgaa agctacatat aaggaacgtg ctgctactca tcctagtcct
     2221 gttgctgcca agctatttaa tatcatgcac gaaaagcaaa caaacttgtg tgcttcattg
     2281 gatgttcgta ccaccaagga attactggag ttagttgaag cattaggtcc caaaatttgt
     2341 ttactaaaaa cacatgtgga tatcttgact gatttttcca tggagggcac agttaagccg
     2401 ctaaaggcat tatccgccaa gtacaatttt ttactcttcg aggacagaaa atttgctgac
     2461 attggtaata cagtcaaatt gcagtactct gcgggtgtat acagaatagc agaatgggca
     2521 gacattacga atgcacacgg tgtggtgggc ccaggtattg ttagcggttt gaagcaggcg
     2581 gcagaagaag taacaaagga acctagaggc cttttgatgt tagcagaatt gtcatgcaag
     2641 ggctccctat ctactggaga atatactaag ggtactgttg acattgcgaa gagcgacaaa
     2701 gattttgtta tcggctttat tgctcaaaga gacatgggtg gaagagatga aggttacgat
     2761 tggttgatta tgacacccgg tgtgggttta gatgacaagg gagatgcatt gggtcaacag
     2821 tatagaaccg tggatgatgt ggtttctaca ggatctgaca ttattattgt tggaagagga
     2881 ctatttgcaa agggaaggga tgctaaggta gagggtgaac gttacagaaa agcaggctgg
     2941 gaagcatatt tgagaagatg cggccagcaa aactaaaaaa ctgtattata agtaaatgca
     3001 tgtatactaa actcacaaat tagagcttca atttaattat atcagttatt acccgagtaa
     3061 cgaagcatct gtgcttcatt ttgtagaaca aaaatgcaac gcgagagcgc taatttttca
     3121 aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgaaag cgctatttta
     3181 ccaacgaaga atctgtgctt catttttgta aaacaaaaat gcaacgcgag agcgctaatt
     3241 tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc gagagcgcta
     3301 ttttaccaac aaagaatcta tacttctttt ttgttctaca aaaatgcatc ccgagagcgc
     3361 tatttttcta acaaagcatc ttagattact ttttttctcc tttgtgcgct ctataatgca
     3421 gtctcttgat aactttttgc actgtaggtc cgttaaggtt agaagaaggc tactttggtg
     3481 tctattttct cttccataaa aaaagcctga ctccacttcc cgcgtttact gattactagc
     3541 gaagctgcgg gtgcattttt tcaagataaa ggcatccccg attatattct ataccgatgt
     3601 ggattgcgca tactttgtga acagaaagtg atagcgttga tgattcttca ttggtcagaa
     3661 aattatgaac ggtttcttct attttgtctc tatatactac gtataggaaa tgtttacatt
     3721 ttcgtattgt tttcgattca ctctatgaat agttcttact acaatttttt tgtctaaaga
     3781 gtaatactag agataaacat aaaaaatgta gaggtcgagt ttagatgcaa gttcaaggag
     3841 cgaaaggtgg atgggtaggt tatataggga tatagcacag agatatatag caaagagata
     3901 cttttgagca atgtttgtgg aagcggtatt cgcaatattt tagtagctcg ttacagtccg
     3961 gtgcgttttt ggttttttga aagtgcgtca tcagagcgct tttggttttc aaaagcgctc
     4021 tgaagttcct atactttcta gctagagaat aggaacttcc cgagcggccg cgtgttacaa
     4081 gcttgactcc agcgtaactg gactgcaatc aactcactgg ctcaccttca cgggtgggcc
     4141 tttcttcggt agaaaatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     4201 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     4261 ccaactcttt ttccgaggta actggcttca gcagagcgca gataccaaat actgttcttc
     4321 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4381 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4441 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4501 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4561 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     4621 gggtcggaac aggagagcgc acgagggagc ttccaggggt aaacgcctgg tatctttata
     4681 gtcctgtcgg gtttcgccac ctctgacttg agcatcgatt tttgtgatgc tcgtcagggg
     4741 ggcggagcct atggaaaaac gccagcaacg cagaaaggcc cacccgaagg tgagccaggt
     4801 gattacattt gggccctcat cagaggtttt caccgtcatc accgaaacgc gcgaggcagc
     4861 tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg
     4921 cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca
     4981 tgttaagggc ggttttttcc tgtttggtca cttaccaatg cttaatcagt gaggcaccta
     5041 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5101 ctacgatgcg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     5161 gctcaccggc tccagattta tcagcaataa accagccagc cgggagggcc gagcgcagaa
     5221 gtgatcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     5281 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     5341 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     5401 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     5461 ccagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     5521 ttactgtcat gccatccgtg agatgctttt ctgtgactgg tgagtactca accaagtcat
     5581 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     5641 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgta
     5701 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     5761 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     5821 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc attttagctt
     5881 ccttagctcc tgaaaatctc gataactcaa aaaatacgcc cggtagtgat cttatttcat
     5941 tatggtgaaa gttggaacct cttacgtgcc gatcaagtca aaagcctccg gtcggaggct
     6001 tttgactttc tgctatggag gtca
//
