LOCUS       sequence-300234-        5484 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 5484)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5484)
  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-m4133747
FEATURES             Location/Qualifiers
     source          1..5484
                     /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..1574
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1627..1748
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1638..1657)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1692..1711
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1863..1909
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1939..2075
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2120..2191
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2417..3005)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2497..2516)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3751..3819,3820..4611)
                     /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(3969..3988)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4624..4760
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4912..4983
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4988..5034)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5052..5100
                     /note="synthetic polyadenylation signal"
     enhancer        5181..5484
                     /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 cagtcgcacc
     1561 atcaccatca ccattaagct agcttgactg actgagatac agcgtacctt cagctcacag
     1621 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     1681 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     1741 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     1801 aggtttttta aagcaagtaa aacctctaca aatgtggtat tggcccatct ctatcggtat
     1861 cgtagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgt gcccctcggg
     1921 ccggattgct atctaccggc attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc
     1981 ttatactccc acatatgcca gattcagcaa cggatacggc ttccccaact tgcccacttc
     2041 catacgtgtc ctccttacca gaaatttatc cttaaggtcg tcagctatcc tgcaggcgat
     2101 ctctcgattt cgatcaagac attcctttaa tggtcttttc tggacaccac taggggtcag
     2161 aagtagttca tcaaactttc ttccctccct aatctcattg gttaccttgg gctatcgaaa
     2221 cttaattaac cagtcaagtc agctacttgg cgagatcgac ttgtctgggt ttcgactacg
     2281 ctcagaattg cgtcagtcaa gttcgatctg gtccttgcta ttgcacccgt tctccgatta
     2341 cgagtttcat ttaaatcatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     2401 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     2461 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     2521 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     2581 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     2641 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     2701 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     2761 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     2821 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     2881 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     2941 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3001 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3061 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3121 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     3181 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     3241 catttaaatt tccgaactct ccaaggccct cgtcggaaaa tcttcaaacc tttcgtccga
     3301 tccatcttgc aggctacctc tcgaacgaac tatcgcaagt ctcttggccg gccttgcgcc
     3361 ttggctattg cttggcagcg cctatcgcca ggtattactc caatcccgaa tatccgagat
     3421 cgggatcacc cgagagaagt tcaacctaca tcctcaatcc cgatctatcc gagatccgag
     3481 gaatatcgaa atcggggcgc gcctggtgta ccgagaacga tcctctcagt gcgagtctcg
     3541 acgatccata tcgttgcttg gcagtcagcc agtcggaatc cagcttggga cccaggaagt
     3601 ccaatcgtca gatattgtac tcaagcctgg tcacggcagc gtaccgatct gtttaaacct
     3661 agatattgat agtctgatcg gtcaacgtat aatcgagtcc tagcttttgc aaacatctat
     3721 caagagacag gatcagcagg aggctttcgc atgagtattc aacatttccg tgtcgccctt
     3781 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     3841 gtaaaagatg ctgaagatca gttgggtgcg cgagtgggtt acatcgaact ggatctcaac
     3901 agcggtaaga tccttgagag ttttcgcccc gaagaacgct ttccaatgat gagcactttt
     3961 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     4021 cgccgcatac actattctca gaatgacttg gttgagtatt caccagtcac agaaaagcat
     4081 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     4141 actgcggcca acttacttct gacaacgatt ggaggaccga aggagctaac cgcttttttg
     4201 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     4261 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac cttgcgtaaa
     4321 ctattaactg gcgaactact tactctagct tcccggcaac agttgataga ctggatggag
     4381 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     4441 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     4501 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     4561 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta accgattcta
     4621 ggtgcattgg cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata
     4681 tgccagattc agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct
     4741 taccagaaat ttatccttaa gatcccgaat cgtttaaact cgactctggc tctatcgaat
     4801 ctccgtcgtt tcgagcttac gcgaacagcc gtggcgctca tttgctcgtc gggcatcgaa
     4861 tctcgtcagc tatcgtcagc ttaccttttt ggcagcgatc gcggctcccg acatcttgga
     4921 ccattagctc cacaggtatc ttcttccctc tagtggtcat aacagcagct tcagctacct
     4981 ctcaattcaa aaaacccctc aagacccgtt tagaggcccc aaggggttat gctatcaatc
     5041 gttgcgttac acacacaaaa aaccaacaca catccatctt cgatggatag cgattttatt
     5101 atctaactgc tgatcgagtg tagccagatc tagtaatcaa ttacggggtc attagttcat
     5161 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
     5221 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
     5281 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     5341 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
     5401 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     5461 gtattagtca tcgctattac catg
//
