LOCUS       sequence-300233-        7860 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 N (nucleocapsid).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7860)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7860)
  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-m4133881
FEATURES             Location/Qualifiers
     source          1..7860
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1536..1556
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1557..1598
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    1667..1788
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1678..1697)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1732..1751
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1903..1949
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1979..2115
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2160..2231
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2457..3045)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2537..2556)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3545..4754
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4591..4610
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4758..5786
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5813..5861
                     /note="synthetic polyadenylation signal"
     CDS             join(6127..6195,6196..6987)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6345..6364)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7000..7136
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7288..7359
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7364..7410)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7428..7476
                     /note="synthetic polyadenylation signal"
     enhancer        7557..7860
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ctgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatcagatc tttgtcgatc
      241 ctaccatcca ctcgacacac ccgccagcgg ccgccaccat gtctgataat ggaccccaaa
      301 atcagcgaaa tgcaccccgc attacgtttg gtggaccctc agattcaact ggcagtaacc
      361 agaatggaga acgcagtggg gcgcgatcaa aacaacgtcg gccccaaggt ttacccaata
      421 atactgcgtc ttggttcacc gctctcactc aacatggcaa ggaagacctt aaattccctc
      481 gaggacaagg cgttccaatt aacaccaata gcagtccaga tgaccaaatt ggctactacc
      541 gaagagctac cagacgaatt cgtggtggtg acggtaaaat gaaagatctc agtccaagat
      601 ggtatttcta ctacctagga actgggccag aagctggact tccctatggt gctaacaaag
      661 acggcatcat atgggttgca actgagggag ccttgaatac accaaaagat cacattggca
      721 cccgcaatcc tgctaacaat gctgcaatcg tgctacaact tcctcaagga acaacattgc
      781 caaaaggctt ctacgcagaa gggagcagag gcggcagtca agcctcttct cgttcctcat
      841 cacgtagtcg caacagttca agaaattcaa ctccaggcag cagtagggga acttctcctg
      901 ctagaatggc tggcaatggc ggtgatgctg ctcttgcttt gctgctgctt gacagattga
      961 accagcttga gagcaaaatg tctggtaaag gccaacaaca acaaggccaa actgtcacta
     1021 agaaatctgc tgctgaggct tctaagaagc ctcggcaaaa acgtactgcc actaaagcat
     1081 acaatgtaac acaagctttc ggcagacgtg gtccagaaca aacccaagga aattttgggg
     1141 accaggaact aatcagacaa ggaactgatt acaaacattg gccgcaaatt gcacaatttg
     1201 cccccagcgc ttcagcgttc ttcggaatgt cgcgcattgg catggaagtc acaccttcgg
     1261 gaacgtggtt gacctacaca ggtgccatca aattggatga caaagatcca aatttcaaag
     1321 atcaagtcat tttgctgaat aagcatattg acgcatacaa aacattccca ccaacagagc
     1381 ctaaaaagga caaaaagaag aaggctgatg aaactcaagc cttaccgcag agacagaaga
     1441 aacagcaaac tgtgactctt cttcctgctg cagatttgga tgatttctcc aaacaattgc
     1501 aacaatccat gagcagtgct gactcaactc aggccgaaaa cctttacttt cagtcgggga
     1561 aacctatacc caatccactt ctcggcttag attctaccta aggatccgca ggcctctgct
     1621 agcttgactg actgagatac agcgtacctt cagctcacag acatgataag atacattgat
     1681 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     1741 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     1801 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa
     1861 aacctctaca aatgtggtat tggcccatct ctatcggtat cgtagcataa ccccttgggg
     1921 cctctaaacg ggtcttgagg ggttttttgt gcccctcggg ccggattgct atctaccggc
     1981 attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc acatatgcca
     2041 gattcagcaa cggatacggc ttccccaact tgcccacttc catacgtgtc ctccttacca
     2101 gaaatttatc cttaaggtcg tcagctatcc tgcaggcgat ctctcgattt cgatcaagac
     2161 attcctttaa tggtcttttc tggacaccac taggggtcag aagtagttca tcaaactttc
     2221 ttccctccct aatctcattg gttaccttgg gctatcgaaa cttaattaac cagtcaagtc
     2281 agctacttgg cgagatcgac ttgtctgggt ttcgactacg ctcagaattg cgtcagtcaa
     2341 gttcgatctg gtccttgcta ttgcacccgt tctccgatta cgagtttcat ttaaatcatg
     2401 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     2461 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     2521 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     2581 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     2641 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     2701 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     2761 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     2821 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     2881 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     2941 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     3001 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     3061 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     3121 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     3181 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     3241 cctatctcag cgatctgtct atttcgttca tccatagttg catttaaatt tccgaactct
     3301 ccaaggccct cgtcggaaaa tcttcaaacc tttcgtccga tccatcttgc aggctacctc
     3361 tcgaacgaac tatcgcaagt ctcttggccg gccttgcgcc ttggctattg cttggcagcg
     3421 cctatcgcca ggtattactc caatcccgaa tatccgagat cgggatcacc cgagagaagt
     3481 tcaacctaca tcctcaatcc cgatctatcc gagatccgag gaatatcgaa atcggggcgc
     3541 gcctggcctc cgcgccgggt tttggcgcct cccgcgggcg cccccctcgt cacggcgagc
     3601 gctgccacgt cagacgaagg gcgcaggagc gtcctgatcc ttccgcccgg acgctcagga
     3661 cagcggcccg ctgctcataa gactcggcct tagaacccca gtatcagcag aaggacattt
     3721 taggacggga cttgggtgac tctagggcac tggttttctt tccagagagc ggaacaggcg
     3781 aggaaaagta gtcccttctc ggcgattctg cggagggatc tccgtggggc ggtgaacgcc
     3841 gatgattata taaggacgcg ccgggtgtgg cacagctagt tccgtcgcag ccgggatttg
     3901 ggtcgcggtt cttgtttgtg gatcgctgtg atcgtcactt ggtgagtagc gggctgctgg
     3961 gctggccggg gctttcgtgg ccgccgggcc gctcggtggg acggaagcgt gtggagagac
     4021 cgccaagggc tgtagtctgg gtccgcgagc aaggttgccc tgaactgggg gttgggggga
     4081 gcgcagcaaa atggcggctg ttcccgagtc ttgaatggaa gacgcttgtg aggcgggctg
     4141 tgaggtcgtt gaaacaaggt ggggggcatg gtgggcggca agaacccaag gtcttgagcc
     4201 cttcgctaat gcgggaaagc tcttattcgg gtgagatggg ctgggcacca tctggggacc
     4261 ctgacgtgaa gtttgtcact gactggagaa ctcggtttgt cgtctgttgc gggggcggca
     4321 gttatggcgg tgccgttggg cagtgcaccc gtacctttgg gagcgcgcgc cctcgtcgtg
     4381 tcgtgacgtc acccgttctg ttggcttata atgcagggtg gggccacctg ccggtaggtg
     4441 tgcggtaggc ttttctccgt cgcaggacgc agggttcggg cctagggtag gctctcctga
     4501 atcgacaggc gccggacctc tggtgagggg agggataagt gaggcgtcag tttctttggt
     4561 cggttttatg tacctatctt cttaagtagc tgaagctccg gttttgaact atgcgctcgg
     4621 ggttggcgag tgtgttttgt gaagtttttt aggcaccttt tgaaatgtaa tcatttgggt
     4681 caatatgtaa ttttcagtgt tagacttgta aattgtccgc taaattctgg ccgtttttgg
     4741 cttttttgtt agacaacatg ggtaaaaagc ctgaactcac cgcgacgtct gtcgagaagt
     4801 ttctgatcga aaagttcgac agcgtctccg acctgatgca gctctcggag ggcgaagaat
     4861 ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt cctgcgggta aatagctgcg
     4921 ccgatggttt ctacaaagat cgttatgttt atcggcactt tgcatccgcc gcgctcccga
     4981 ttccggaagt gcttgacatt ggggagttca gcgagagcct gacctattgc atctcccgcc
     5041 gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga actgcccgct gttctgcagc
     5101 cggtcgcgga ggcaatggat gccatcgctg ccgccgatct tagccagacg agcgggttcg
     5161 gcccattcgg accgcaagga atcggtcaat acactacatg gcgtgatttc atatgcgcga
     5221 ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg
     5281 tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga ctgccccgaa gtccggcacc
     5341 tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga caatggccgc ataacagcgg
     5401 tcattgactg gagcgaggcg atgttcgggg attcccaata cgaggtcgcc aacatcttct
     5461 tctggaggcc gtggttggct tgtatggagc agcagacgcg ctacttcgag cggaggcatc
     5521 cggagcttgc aggatcgccc cggctccggg cgtatatgct ccgcattggt cttgaccaac
     5581 tctatcagag cttggttgac ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg
     5641 acgcaatcgt ccgatccgga gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg
     5701 ccgccgtctg gaccgatggc tgtgtagaag tactcgccga tagtggaaac cgacgcccca
     5761 gcactcgtcc gagggcaaag gaataagcta gtatgtaagc ctagtcttag ataataaaat
     5821 cgctatccat cgaagatgga tgtgtgttgg ttttttgtgt gtgtaacgct aggcgcgcct
     5881 ggtgtaccga gaacgatcct ctcagtgcga gtctcgacga tccatatcgt tgcttggcag
     5941 tcagccagtc ggaatccagc ttgggaccca ggaagtccaa tcgtcagata ttgtactcaa
     6001 gcctggtcac ggcagcgtac cgatctgttt aaacctagat attgatagtc tgatcggtca
     6061 acgtataatc gagtcctagc ttttgcaaac atctatcaag agacaggatc agcaggaggc
     6121 tttcgcatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     6181 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     6241 ggtgcgcgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     6301 cgccccgaag aacgctttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     6361 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     6421 gacttggttg agtattcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     6481 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     6541 acgattggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     6601 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     6661 acgatgcctg tagcaatggc aacaaccttg cgtaaactat taactggcga actacttact
     6721 ctagcttccc ggcaacagtt gatagactgg atggaggcgg ataaagttgc aggaccactt
     6781 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     6841 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     6901 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     6961 ggtgcctcac tgattaagca ttggtaaccg attctaggtg cattggcgca gaaaaaaatg
     7021 cctgatgcga cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg
     7081 cttccccaac ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaagatc
     7141 ccgaatcgtt taaactcgac tctggctcta tcgaatctcc gtcgtttcga gcttacgcga
     7201 acagccgtgg cgctcatttg ctcgtcgggc atcgaatctc gtcagctatc gtcagcttac
     7261 ctttttggca gcgatcgcgg ctcccgacat cttggaccat tagctccaca ggtatcttct
     7321 tccctctagt ggtcataaca gcagcttcag ctacctctca attcaaaaaa cccctcaaga
     7381 cccgtttaga ggccccaagg ggttatgcta tcaatcgttg cgttacacac acaaaaaacc
     7441 aacacacatc catcttcgat ggatagcgat tttattatct aactgctgat cgagtgtagc
     7501 cagatctagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     7561 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     7621 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     7681 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     7741 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     7801 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
//
