LOCUS       sequence-307256-        7451 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 7451)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7451)
  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-m4133812
FEATURES             Location/Qualifiers
     source          1..7451
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    18..66
                     /note="synthetic polyadenylation signal"
     enhancer        147..450
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        452..654
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     605..625
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1986..2006
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2007..2084
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     polyA_signal    2137..2258
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2148..2167)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2202..2221
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2373..2419
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2449..2585
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2630..2701
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2927..3515)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3007..3026)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4015..5224
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5061..5080
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5228..5827
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5228..5247)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5724..5744
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5854..5902
                     /note="synthetic polyadenylation signal"
     CDS             join(6168..6236,6237..7028)
                     /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(6386..6405)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7041..7177
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7329..7400
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7405..7451)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 tcaatcgttg cgttacacac acaaaaaacc aacacacatc catcttcgat ggatagcgat
       61 tttattatct aactgctgat cgagtgtagc cagatctagt aatcaattac ggggtcatta
      121 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
      181 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      241 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      301 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      361 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      421 atctacgtat tagtcatcgc tattaccatg ctgatgcggt tttggcagta catcaatggg
      481 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      541 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      601 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta
      661 gtgaaccgtc agatcagatc tttgtcgatc ctaccatcca ctcgacacac ccgccagcgg
      721 ccgccaccat gtctgataat ggaccccaaa atcagcgaaa tgcaccccgc attacgtttg
      781 gtggaccctc agattcaact ggcagtaacc agaatggaga acgcagtggg gcgcgatcaa
      841 aacaacgtcg gccccaaggt ttacccaata atactgcgtc ttggttcacc gctctcactc
      901 aacatggcaa ggaagacctt aaattccctc gaggacaagg cgttccaatt aacaccaata
      961 gcagtccaga tgaccaaatt ggctactacc gaagagctac cagacgaatt cgtggtggtg
     1021 acggtaaaat gaaagatctc agtccaagat ggtatttcta ctacctagga actgggccag
     1081 aagctggact tccctatggt gctaacaaag acggcatcat atgggttgca actgagggag
     1141 ccttgaatac accaaaagat cacattggca cccgcaatcc tgctaacaat gctgcaatcg
     1201 tgctacaact tcctcaagga acaacattgc caaaaggctt ctacgcagaa gggagcagag
     1261 gcggcagtca agcctcttct cgttcctcat cacgtagtcg caacagttca agaaattcaa
     1321 ctccaggcag cagtagggga acttctcctg ctagaatggc tggcaatggc ggtgatgctg
     1381 ctcttgcttt gctgctgctt gacagattga accagcttga gagcaaaatg tctggtaaag
     1441 gccaacaaca acaaggccaa actgtcacta agaaatctgc tgctgaggct tctaagaagc
     1501 ctcggcaaaa acgtactgcc actaaagcat acaatgtaac acaagctttc ggcagacgtg
     1561 gtccagaaca aacccaagga aattttgggg accaggaact aatcagacaa ggaactgatt
     1621 acaaacattg gccgcaaatt gcacaatttg cccccagcgc ttcagcgttc ttcggaatgt
     1681 cgcgcattgg catggaagtc acaccttcgg gaacgtggtt gacctacaca ggtgccatca
     1741 aattggatga caaagatcca aatttcaaag atcaagtcat tttgctgaat aagcatattg
     1801 acgcatacaa aacattccca ccaacagagc ctaaaaagga caaaaagaag aaggctgatg
     1861 aaactcaagc cttaccgcag agacagaaga aacagcaaac tgtgactctt cttcctgctg
     1921 cagatttgga tgatttctcc aaacaattgc aacaatccat gagcagtgct gactcaactc
     1981 aggccgaaaa cctttacttt cagtcgaaga ggcgatggaa aaagaatttt atcgcggtga
     2041 gtgcagccaa cagattcaaa aagatatcct cgtctggtgc cctctaagct agcttgactg
     2101 actgagatac agcgtacctt cagctcacag acatgataag atacattgat gagtttggac
     2161 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     2221 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     2281 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
     2341 aatgtggtat tggcccatct ctatcggtat cgtagcataa ccccttgggg cctctaaacg
     2401 ggtcttgagg ggttttttgt gcccctcggg ccggattgct atctaccggc attggcgcag
     2461 aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa
     2521 cggatacggc ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc
     2581 cttaaggtcg tcagctatcc tgcaggcgat ctctcgattt cgatcaagac attcctttaa
     2641 tggtcttttc tggacaccac taggggtcag aagtagttca tcaaactttc ttccctccct
     2701 aatctcattg gttaccttgg gctatcgaaa cttaattaac cagtcaagtc agctacttgg
     2761 cgagatcgac ttgtctgggt ttcgactacg ctcagaattg cgtcagtcaa gttcgatctg
     2821 gtccttgcta ttgcacccgt tctccgatta cgagtttcat ttaaatcatg tgagcaaaag
     2881 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     2941 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     3001 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     3061 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     3121 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     3181 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3241 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     3301 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     3361 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     3421 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     3481 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     3541 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     3601 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     3661 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     3721 cgatctgtct atttcgttca tccatagttg catttaaatt tccgaactct ccaaggccct
     3781 cgtcggaaaa tcttcaaacc tttcgtccga tccatcttgc aggctacctc tcgaacgaac
     3841 tatcgcaagt ctcttggccg gccttgcgcc ttggctattg cttggcagcg cctatcgcca
     3901 ggtattactc caatcccgaa tatccgagat cgggatcacc cgagagaagt tcaacctaca
     3961 tcctcaatcc cgatctatcc gagatccgag gaatatcgaa atcggggcgc gcctggcctc
     4021 cgcgccgggt tttggcgcct cccgcgggcg cccccctcgt cacggcgagc gctgccacgt
     4081 cagacgaagg gcgcaggagc gtcctgatcc ttccgcccgg acgctcagga cagcggcccg
     4141 ctgctcataa gactcggcct tagaacccca gtatcagcag aaggacattt taggacggga
     4201 cttgggtgac tctagggcac tggttttctt tccagagagc ggaacaggcg aggaaaagta
     4261 gtcccttctc ggcgattctg cggagggatc tccgtggggc ggtgaacgcc gatgattata
     4321 taaggacgcg ccgggtgtgg cacagctagt tccgtcgcag ccgggatttg ggtcgcggtt
     4381 cttgtttgtg gatcgctgtg atcgtcactt ggtgagtagc gggctgctgg gctggccggg
     4441 gctttcgtgg ccgccgggcc gctcggtggg acggaagcgt gtggagagac cgccaagggc
     4501 tgtagtctgg gtccgcgagc aaggttgccc tgaactgggg gttgggggga gcgcagcaaa
     4561 atggcggctg ttcccgagtc ttgaatggaa gacgcttgtg aggcgggctg tgaggtcgtt
     4621 gaaacaaggt ggggggcatg gtgggcggca agaacccaag gtcttgagcc cttcgctaat
     4681 gcgggaaagc tcttattcgg gtgagatggg ctgggcacca tctggggacc ctgacgtgaa
     4741 gtttgtcact gactggagaa ctcggtttgt cgtctgttgc gggggcggca gttatggcgg
     4801 tgccgttggg cagtgcaccc gtacctttgg gagcgcgcgc cctcgtcgtg tcgtgacgtc
     4861 acccgttctg ttggcttata atgcagggtg gggccacctg ccggtaggtg tgcggtaggc
     4921 ttttctccgt cgcaggacgc agggttcggg cctagggtag gctctcctga atcgacaggc
     4981 gccggacctc tggtgagggg agggataagt gaggcgtcag tttctttggt cggttttatg
     5041 tacctatctt cttaagtagc tgaagctccg gttttgaact atgcgctcgg ggttggcgag
     5101 tgtgttttgt gaagtttttt aggcaccttt tgaaatgtaa tcatttgggt caatatgtaa
     5161 ttttcagtgt tagacttgta aattgtccgc taaattctgg ccgtttttgg cttttttgtt
     5221 agacaacatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag
     5281 ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga
     5341 tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg
     5401 gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac
     5461 gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt
     5521 gagcggttcc cggctggccg cgcagcaaca gatggagggc ctcctggcgc cgcaccggcc
     5581 caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg
     5641 tctgggcagc gccgtcgtgc tccccggagt ggaggccgcc gagcgcgccg gggtgcccgc
     5701 cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt
     5761 caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg
     5821 tgcctaggct agtatgtaag cctagtctta gataataaaa tcgctatcca tcgaagatgg
     5881 atgtgtgttg gttttttgtg tgtgtaacgc taggcgcgcc tggtgtaccg agaacgatcc
     5941 tctcagtgcg agtctcgacg atccatatcg ttgcttggca gtcagccagt cggaatccag
     6001 cttgggaccc aggaagtcca atcgtcagat attgtactca agcctggtca cggcagcgta
     6061 ccgatctgtt taaacctaga tattgatagt ctgatcggtc aacgtataat cgagtcctag
     6121 cttttgcaaa catctatcaa gagacaggat cagcaggagg ctttcgcatg agtattcaac
     6181 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     6241 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcgcga gtgggttaca
     6301 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgctttc
     6361 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     6421 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtattcac
     6481 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     6541 taaccatgag tgataacact gcggccaact tacttctgac aacgattgga ggaccgaagg
     6601 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     6661 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     6721 caacaacctt gcgtaaacta ttaactggcg aactacttac tctagcttcc cggcaacagt
     6781 tgatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     6841 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     6901 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     6961 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     7021 attggtaacc gattctaggt gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg
     7081 tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact
     7141 tccatacgtg tcctccttac cagaaattta tccttaagat cccgaatcgt ttaaactcga
     7201 ctctggctct atcgaatctc cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt
     7261 gctcgtcggg catcgaatct cgtcagctat cgtcagctta cctttttggc agcgatcgcg
     7321 gctcccgaca tcttggacca ttagctccac aggtatcttc ttccctctag tggtcataac
     7381 agcagcttca gctacctctc aattcaaaaa acccctcaag acccgtttag aggccccaag
     7441 gggttatgct a
//
