LOCUS       sequence-300327-        5544 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 5544)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5544)
  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-m4134480
FEATURES             Location/Qualifiers
     source          1..5544
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    97..168
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(173..219)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    237..285
                     /note="synthetic polyadenylation signal"
     enhancer        366..669
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        671..873
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     824..844
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2205..2225
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2226..2303
                     /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    2356..2477
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2367..2386)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2421..2440
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2592..2638
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2668..2804
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2849..2920
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3146..3734)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3226..3245)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(4480..4548,4549..5340)
                     /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(4698..4717)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5353..5489
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 cttacgcgaa cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg
       61 tcagcttacc tttttggcag cgatcgcggc tcccgacatc ttggaccatt agctccacag
      121 gtatcttctt ccctctagtg gtcataacag cagcttcagc tacctctcaa ttcaaaaaac
      181 ccctcaagac ccgtttagag gccccaaggg gttatgctat caatcgttgc gttacacaca
      241 caaaaaacca acacacatcc atcttcgatg gatagcgatt ttattatcta actgctgatc
      301 gagtgtagcc agatctagta atcaattacg gggtcattag ttcatagccc atatatggag
      361 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      421 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      481 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      541 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      601 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      661 attaccatgc tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      721 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      781 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      841 cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca gatcagatct
      901 ttgtcgatcc taccatccac tcgacacacc cgccagcggc cgccaccatg tctgataatg
      961 gaccacagaa ccaacgcaat gcaccccgaa tcacttttgg cggtccgtcc gacagtaccg
     1021 ggagcaacca gaacggcgaa agatcgggag ctcgttcaaa acagcggagg cctcaagggt
     1081 tacccaataa cacggccagc tggttcacag cgttgaccca gcatggcaag gaggacctca
     1141 agttccctcg gggtcagggg gtcccaataa atacaaacag ttcccctgat gaccaaattg
     1201 gatactatag acgcgctact aggaggatcc gtggcgggga tggaaaaatg aaggaccttt
     1261 ctcccagatg gtacttttat tacctgggta ctggcccaga ggccggtcta ccgtatggag
     1321 caaacaaaga tggcattatt tgggtggcca cggaaggagc actgaatacc ccaaaggatc
     1381 acatcggcac ccggaatcca gccaacaatg ctgctatcgt tctgcaactc ccccagggga
     1441 caacattgcc taaaggcttc tatgccgagg gcagtagagg tggcagccag gcgtcctcac
     1501 gcagctcttc tcgatctcgg aacagctcaa gaaattccac tcccgggagt tcccgaggaa
     1561 ccagcccagc aaggatggct ggcaatggtg gagatgccgc tctggcccta cttctgctgg
     1621 accgcctcaa tcagttagaa tcaaagatgt cggggaaggg gcagcaacag cagggacaga
     1681 ccgtaactaa aaagagtgca gcggaggcct ctaaaaagcc tcggcagaaa cgtacggcaa
     1741 caaaggctta caacgtgacc caggcctttg gccgtagagg acccgaacag acacaaggga
     1801 actttggtga ccaagagttg ataaggcagg gaaccgacta caaacactgg cctcagatcg
     1861 cccaattcgc accgagcgcg agcgcatttt tcggaatgtc acgaattggc atggaggtga
     1921 caccttcggg aacctggctg acatacactg gcgccataaa gctcgatgac aaagatccca
     1981 acttcaagga tcaagtcatc ttgctgaata agcatattga cgcgtataaa acttttccac
     2041 ccacggaacc gaagaaggac aagaaaaaga aagctgatga gacacaggcg cttccacaaa
     2101 ggcagaaaaa gcagcagacg gttactctac tgcccgccgc agacctagat gacttctcca
     2161 aacagcttca gcagtctatg agtagcgccg actcaaccca agctgaaaac ctttactttc
     2221 agtcgaagag gcgatggaaa aagaatttta tcgcggtgag tgcagccaac agattcaaaa
     2281 agatatcctc gtctggtgcc ctctaagcta gcttgactga ctgagataca gcgtaccttc
     2341 agctcacaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     2401 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     2461 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     2521 aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatt ggcccatctc
     2581 tatcggtatc gtagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgtg
     2641 cccctcgggc cggattgcta tctaccggca ttggcgcaga aaaaaatgcc tgatgcgacg
     2701 ctgcgcgtct tatactccca catatgccag attcagcaac ggatacggct tccccaactt
     2761 gcccacttcc atacgtgtcc tccttaccag aaatttatcc ttaaggtcgt cagctatcct
     2821 gcaggcgatc tctcgatttc gatcaagaca ttcctttaat ggtcttttct ggacaccact
     2881 aggggtcaga agtagttcat caaactttct tccctcccta atctcattgg ttaccttggg
     2941 ctatcgaaac ttaattaacc agtcaagtca gctacttggc gagatcgact tgtctgggtt
     3001 tcgactacgc tcagaattgc gtcagtcaag ttcgatctgg tccttgctat tgcacccgtt
     3061 ctccgattac gagtttcatt taaatcatgt gagcaaaagg ccagcaaaag gccaggaacc
     3121 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     3181 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     3241 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     3301 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     3361 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     3421 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     3481 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     3541 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     3601 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     3661 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     3721 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     3781 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     3841 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     3901 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     3961 ccatagttgc atttaaattt ccgaactctc caaggccctc gtcggaaaat cttcaaacct
     4021 ttcgtccgat ccatcttgca ggctacctct cgaacgaact atcgcaagtc tcttggccgg
     4081 ccttgcgcct tggctattgc ttggcagcgc ctatcgccag gtattactcc aatcccgaat
     4141 atccgagatc gggatcaccc gagagaagtt caacctacat cctcaatccc gatctatccg
     4201 agatccgagg aatatcgaaa tcggggcgcg cctggtgtac cgagaacgat cctctcagtg
     4261 cgagtctcga cgatccatat cgttgcttgg cagtcagcca gtcggaatcc agcttgggac
     4321 ccaggaagtc caatcgtcag atattgtact caagcctggt cacggcagcg taccgatctg
     4381 tttaaaccta gatattgata gtctgatcgg tcaacgtata atcgagtcct agcttttgca
     4441 aacatctatc aagagacagg atcagcagga ggctttcgca tgagtattca acatttccgt
     4501 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     4561 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcgc gagtgggtta catcgaactg
     4621 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgctt tccaatgatg
     4681 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     4741 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtattc accagtcaca
     4801 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     4861 agtgataaca ctgcggccaa cttacttctg acaacgattg gaggaccgaa ggagctaacc
     4921 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     4981 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacc
     5041 ttgcgtaaac tattaactgg cgaactactt actctagctt cccggcaaca gttgatagac
     5101 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     5161 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     5221 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     5281 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     5341 ccgattctag gtgcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac
     5401 tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg
     5461 tgtcctcctt accagaaatt tatccttaag atcccgaatc gtttaaactc gactctggct
     5521 ctatcgaatc tccgtcgttt cgag
//
