LOCUS       sequence-287604-        6013 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 6013)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6013)
  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-m4046174
FEATURES             Location/Qualifiers
     source          1..6013
                     /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             1688..1705
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1706..1723
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1768..1786)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2137..2940
                     /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      3023..4005
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4083..4112
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4134..4721
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4733..4760)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4784..4975)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4976..5767,5768..5836))
                     /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     5599..5618
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5837..5939)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5940..5971)
                     /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 atggacccca aaatcagcga aatgcacccc gcattacgtt
      481 tggtggaccc tcagattcaa ctggcagtaa ccagaatgga gaacgcagtg gggcgcgatc
      541 aaaacaacgt cggccccaag gtttacccaa taatactgcg tcttggttca ccgctctcac
      601 tcaacatggc aaggaagacc ttaaattccc tcgaggacaa ggcgttccaa ttaacaccaa
      661 tagcagtcca gatgaccaaa ttggctacta ccgaagagct accagacgaa ttcgtggtgg
      721 tgacggtaaa atgaaagatc tcagtccaag atggtatttc tactacctag gaactgggcc
      781 agaagctgga cttccctatg gtgctaacaa agacggcatc atatgggttg caactgaggg
      841 agccttgaat acaccaaaag atcacattgg cacccgcaat cctgctaaca atgctgcaat
      901 cgtgctacaa cttcctcaag gaacaacatt gccaaaaggc ttctacgcag aagggagcag
      961 aggcggcagt caagcctctt ctcgttcctc atcacgtagt cgcaacagtt caagaaattc
     1021 aactccaggc agcagtaggg gaacttctcc tgctagaatg gctggcaatg gcggtgatgc
     1081 tgctcttgct ttgctgctgc ttgacagatt gaaccagctt gagagcaaaa tgtctggtaa
     1141 aggccaacaa caacaaggcc aaactgtcac taagaaatct gctgctgagg cttctaagaa
     1201 gcctcggcaa aaacgtactg ccactaaagc atacaatgta acacaagctt tcggcagacg
     1261 tggtccagaa caaacccaag gaaattttgg ggaccaggaa ctaatcagac aaggaactga
     1321 ttacaaacat tggccgcaaa ttgcacaatt tgcccccagc gcttcagcgt tcttcggaat
     1381 gtcgcgcatt ggcatggaag tcacaccttc gggaacgtgg ttgacctaca caggtgccat
     1441 caaattggat gacaaagatc caaatttcaa agatcaagtc attttgctga ataagcatat
     1501 tgacgcatac aaaacattcc caccaacaga gcctaaaaag gacaaaaaga agaaggctga
     1561 tgaaactcaa gccttaccgc agagacagaa gaaacagcaa actgtgactc ttcttcctgc
     1621 tgcagatttg gatgatttct ccaaacaatt gcaacaatcc atgagcagtg ctgactcaac
     1681 tcaggccctc gtgccgagag gttctcacca tcatcaccat cattaaacag gccccttttc
     1741 ctttgtcgat atcatgtaat tagttatgtc acgcttacat tcacgccctc ctcccacatc
     1801 cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt
     1861 ttaatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg
     1921 tacaaagcgg ccgccgaacc actagtgcac tgcagtacac ggtttccttg aaattttttt
     1981 gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg agcacagact tagattggta
     2041 tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt aacccaactg
     2101 cacagaacaa aaacgtgcag gaaacgaaga taaatcatgt cgaaagctac atataaggaa
     2161 cgtgctgcta ctcatcctag tcctgttgct gccaagctat ttaatatcat gcacgaaaag
     2221 caaacaaact tgtgtgcttc attggatgtt cgtaccacca aggaattact ggagttagtt
     2281 gaagcattag gtcccaaaat ttgtttacta aaaacacatg tggatatctt gactgatttt
     2341 tccatggagg gcacagttaa gccgctaaag gcattatccg ccaagtacaa ttttttactc
     2401 ttcgaggaca gaaaatttgc tgacattggt aatacagtca aattgcagta ctctgcgggt
     2461 gtatacagaa tagcagaatg ggcagacatt acgaatgcac acggtgtggt gggcccaggt
     2521 attgttagcg gtttgaagca ggcggcagaa gaagtaacaa aggaacctag aggccttttg
     2581 atgttagcag aattgtcatg caagggctcc ctatctactg gagaatatac taagggtact
     2641 gttgacattg cgaagagcga caaagatttt gttatcggct ttattgctca aagagacatg
     2701 ggtggaagag atgaaggtta cgattggttg attatgacac ccggtgtggg tttagatgac
     2761 aagggagatg cattgggtca acagtataga accgtggatg atgtggtttc tacaggatct
     2821 gacattatta ttgttggaag aggactattt gcaaagggaa gggatgctaa ggtagagggt
     2881 gaacgttaca gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa
     2941 aaaactgtat tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa
     3001 ttatatcagt tattacccga gtaacgaagc atctgtgctt cattttgtag aacaaaaatg
     3061 caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa
     3121 atgcaacgcg aaagcgctat tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa
     3181 aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa
     3241 cagaaatgca acgcgagagc gctattttac caacaaagaa tctatacttc ttttttgttc
     3301 tacaaaaatg catcccgaga gcgctatttt tctaacaaag catcttagat tacttttttt
     3361 ctcctttgtg cgctctataa tgcagtctct tgataacttt ttgcactgta ggtccgttaa
     3421 ggttagaaga aggctacttt ggtgtctatt ttctcttcca taaaaaaagc ctgactccac
     3481 ttcccgcgtt tactgattac tagcgaagct gcgggtgcat tttttcaaga taaaggcatc
     3541 cccgattata ttctataccg atgtggattg cgcatacttt gtgaacagaa agtgatagcg
     3601 ttgatgattc ttcattggtc agaaaattat gaacggtttc ttctattttg tctctatata
     3661 ctacgtatag gaaatgttta cattttcgta ttgttttcga ttcactctat gaatagttct
     3721 tactacaatt tttttgtcta aagagtaata ctagagataa acataaaaaa tgtagaggtc
     3781 gagtttagat gcaagttcaa ggagcgaaag gtggatgggt aggttatata gggatatagc
     3841 acagagatat atagcaaaga gatacttttg agcaatgttt gtggaagcgg tattcgcaat
     3901 attttagtag ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc gtcatcagag
     3961 cgcttttggt tttcaaaagc gctctgaagt tcctatactt tctagctaga gaataggaac
     4021 ttcccgagcg gccgcgtgtt acaagcttga ctccagcgta actggactgc aatcaactca
     4081 ctggctcacc ttcacgggtg ggcctttctt cggtagaaaa tcaaaggatc ttcttgagat
     4141 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     4201 gtttgtttgc cggatcaaga gctaccaact ctttttccga ggtaactggc ttcagcagag
     4261 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     4321 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     4381 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     4441 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     4501 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     4561 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     4621 gggtaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcatc
     4681 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcagaaa
     4741 ggcccacccg aaggtgagcc aggtgattac atttgggccc tcatcagagg ttttcaccgt
     4801 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt
     4861 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg
     4921 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcacttacc
     4981 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     5041 cctgactccc cgtcgtgtag ataactacga tgcgggaggg cttaccatct ggccccagtg
     5101 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     5161 cagccgggag ggccgagcgc agaagtgatc ctgcaacttt atccgcctcc atccagtcta
     5221 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     5281 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     5341 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     5401 gctccttcgg tcctccgatc gttgccagaa gtaagttggc cgcagtgtta tcactcatgg
     5461 ttatggcagc actgcataat tctcttactg tcatgccatc cgtgagatgc ttttctgtga
     5521 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     5581 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     5641 ttggaaaacg ttcttcgggg cgtaaactct caaggatctt accgctgttg agatccagtt
     5701 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     5761 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     5821 aatgttgaat actcatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata
     5881 cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa
     5941 gtcaaaagcc tccggtcgga ggcttttgac tttctgctat ggaggtcagg tatgatttaa
     6001 atggtcagtt aga
//
