LOCUS       sequence-295673-        6021 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 6021)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6021)
  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-m4134544
FEATURES             Location/Qualifiers
     source          1..6021
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1262..1282
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1316..1393
                     /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"
     CDS             1421..1441
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    1890..2011
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1901..1920)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1955..1974
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2126..2172
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2202..2338
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2383..2454
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2680..3268)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2760..2779)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(4014..4082,4083..4874)
                     /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(4232..4251)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4887..5023
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5175..5246
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5251..5297)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5315..5363
                     /note="synthetic polyadenylation signal"
     enhancer        5444..5747
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5749..5951
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5902..5922
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgtct gataatggac cacagaacca acgcaatgca ccccgaatca cttttggcgg
       61 tccgtccgac agtaccggga gcaaccagaa cggcgaaaga tcgggagctc gttcaaaaca
      121 gcggaggcct caagggttac ccaataacac ggccagctgg ttcacagcgt tgacccagca
      181 tggcaaggag gacctcaagt tccctcgggg tcagggggtc ccaataaata caaacagttc
      241 ccctgatgac caaattggat actatagacg cgctactagg aggatccgtg gcggggatgg
      301 aaaaatgaag gacctttctc ccagatggta cttttattac ctgggtactg gcccagaggc
      361 cggtctaccg tatggagcaa acaaagatgg cattatttgg gtggccacgg aaggagcact
      421 gaatacccca aaggatcaca tcggcacccg gaatccagcc aacaatgctg ctatcgttct
      481 gcaactcccc caggggacaa cattgcctaa aggcttctat gccgagggca gtagaggtgg
      541 cagccaggcg tcctcacgca gctcttctcg atctcggaac agctcaagaa attccactcc
      601 cgggagttcc cgaggaacca gcccagcaag gatggctggc aatggtggag atgccgctct
      661 ggccctactt ctgctggacc gcctcaatca gttagaatca aagatgtcgg ggaaggggca
      721 gcaacagcag ggacagaccg taactaaaaa gagtgcagcg gaggcctcta aaaagcctcg
      781 gcagaaacgt acggcaacaa aggcttacaa cgtgacccag gcctttggcc gtagaggacc
      841 cgaacagaca caagggaact ttggtgacca agagttgata aggcagggaa ccgactacaa
      901 acactggcct cagatcgccc aattcgcacc gagcgcgagc gcatttttcg gaatgtcacg
      961 aattggcatg gaggtgacac cttcgggaac ctggctgaca tacactggcg ccataaagct
     1021 cgatgacaaa gatcccaact tcaaggatca agtcatcttg ctgaataagc atattgacgc
     1081 gtataaaact tttccaccca cggaaccgaa gaaggacaag aaaaagaaag ctgatgagac
     1141 acaggcgctt ccacaaaggc agaaaaagca gcagacggtt actctactgc ccgccgcaga
     1201 cctagatgac ttctccaaac agcttcagca gtctatgagt agcgccgact caacccaagc
     1261 tgaaaacctt tactttcagt cgggacggcg aatcccgggg ctgataaatc cctggaaaag
     1321 gagatggaag aagaacttta ttgcagtgtc tgcggctaac cggttcaaaa agatcagttc
     1381 aagcggcgcc ctagattacg acattcctac gaccgcctcc gagaatctct atttccaggg
     1441 tgaatttggc ttggcacaac atgacgaggc cgtcgataac aagtttaata aagaacagca
     1501 gaacgctttc tatgagatcc ttcacttacc aaatctgaac gaagagcaac gcaacgcttt
     1561 cattcagagc ctgaaagacg atccctcgca gtctgcaaat ctcctggccg aagccaagaa
     1621 gctgaatgac gcacaggctc caaaagttga taacaagttt aacaaggagc agcagaatgc
     1681 tttctacgag atattgcacc ttcctaatct gaatgaggaa caaagaaatg catttatcca
     1741 gtccctaaaa gacgatccat cacagagtgc caacctgctt gcggaagcca aaaagctcaa
     1801 tgatgcccag gccccgaaag tagacgcgaa ccatcaataa gctagcttga ctgactgaga
     1861 tacagcgtac cttcagctca cagacatgat aagatacatt gatgagtttg gacaaaccac
     1921 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     1981 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     2041 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     2101 tattggccca tctctatcgg tatcgtagca taaccccttg gggcctctaa acgggtcttg
     2161 aggggttttt tgtgcccctc gggccggatt gctatctacc ggcattggcg cagaaaaaaa
     2221 tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac
     2281 ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaagg
     2341 tcgtcagcta tcctgcaggc gatctctcga tttcgatcaa gacattcctt taatggtctt
     2401 ttctggacac cactaggggt cagaagtagt tcatcaaact ttcttccctc cctaatctca
     2461 ttggttacct tgggctatcg aaacttaatt aaccagtcaa gtcagctact tggcgagatc
     2521 gacttgtctg ggtttcgact acgctcagaa ttgcgtcagt caagttcgat ctggtccttg
     2581 ctattgcacc cgttctccga ttacgagttt catttaaatc atgtgagcaa aaggccagca
     2641 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     2701 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2761 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2821 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     2881 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     2941 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3001 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3061 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3121 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3181 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3241 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     3301 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     3361 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     3421 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     3481 tctatttcgt tcatccatag ttgcatttaa atttccgaac tctccaaggc cctcgtcgga
     3541 aaatcttcaa acctttcgtc cgatccatct tgcaggctac ctctcgaacg aactatcgca
     3601 agtctcttgg ccggccttgc gccttggcta ttgcttggca gcgcctatcg ccaggtatta
     3661 ctccaatccc gaatatccga gatcgggatc acccgagaga agttcaacct acatcctcaa
     3721 tcccgatcta tccgagatcc gaggaatatc gaaatcgggg cgcgcctggt gtaccgagaa
     3781 cgatcctctc agtgcgagtc tcgacgatcc atatcgttgc ttggcagtca gccagtcgga
     3841 atccagcttg ggacccagga agtccaatcg tcagatattg tactcaagcc tggtcacggc
     3901 agcgtaccga tctgtttaaa cctagatatt gatagtctga tcggtcaacg tataatcgag
     3961 tcctagcttt tgcaaacatc tatcaagaga caggatcagc aggaggcttt cgcatgagta
     4021 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     4081 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcgcgagtgg
     4141 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     4201 gctttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     4261 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     4321 attcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     4381 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg attggaggac
     4441 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     4501 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     4561 caatggcaac aaccttgcgt aaactattaa ctggcgaact acttactcta gcttcccggc
     4621 aacagttgat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     4681 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     4741 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     4801 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     4861 ttaagcattg gtaaccgatt ctaggtgcat tggcgcagaa aaaaatgcct gatgcgacgc
     4921 tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg
     4981 cccacttcca tacgtgtcct ccttaccaga aatttatcct taagatcccg aatcgtttaa
     5041 actcgactct ggctctatcg aatctccgtc gtttcgagct tacgcgaaca gccgtggcgc
     5101 tcatttgctc gtcgggcatc gaatctcgtc agctatcgtc agcttacctt tttggcagcg
     5161 atcgcggctc ccgacatctt ggaccattag ctccacaggt atcttcttcc ctctagtggt
     5221 cataacagca gcttcagcta cctctcaatt caaaaaaccc ctcaagaccc gtttagaggc
     5281 cccaaggggt tatgctatca atcgttgcgt tacacacaca aaaaaccaac acacatccat
     5341 cttcgatgga tagcgatttt attatctaac tgctgatcga gtgtagccag atctagtaat
     5401 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
     5461 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     5521 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     5581 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
     5641 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     5701 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatgctg atgcggtttt
     5761 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     5821 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     5881 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     5941 taagcagagc tggtttagtg aaccgtcaga tcagatcttt gtcgatccta ccatccactc
     6001 gacacacccg ccagcggccg c
//
