LOCUS       sequence-312892-        5535 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 5535)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5535)
  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-m4134524
FEATURES             Location/Qualifiers
     source          1..5535
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1840..1860
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1864..1929
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1982..2103
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1993..2012)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2047..2066
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2218..2264
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2294..2430
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2475..2546
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2772..3360)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2852..2871)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(4106..4174,4175..4966)
                     /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(4324..4343)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4979..5115
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5267..5338
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5343..5389)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5407..5455
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
      541 gatcctacca tccactcgac acacccgcca gcggccgcca ccatgtctga taatggacca
      601 cagaaccaac gcaatgcacc ccgaatcact tttggcggtc cgtccgacag taccgggagc
      661 aaccagaacg gcgaaagatc gggagctcgt tcaaaacagc ggaggcctca agggttaccc
      721 aataacacgg ccagctggtt cacagcgttg acccagcatg gcaaggagga cctcaagttc
      781 cctcggggtc agggggtccc aataaataca aacagttccc ctgatgacca aattggatac
      841 tatagacgcg ctactaggag gatccgtggc ggggatggaa aaatgaagga cctttctccc
      901 agatggtact tttattacct gggtactggc ccagaggccg gtctaccgta tggagcaaac
      961 aaagatggca ttatttgggt ggccacggaa ggagcactga ataccccaaa ggatcacatc
     1021 ggcacccgga atccagccaa caatgctgct atcgttctgc aactccccca ggggacaaca
     1081 ttgcctaaag gcttctatgc cgagggcagt agaggtggca gccaggcgtc ctcacgcagc
     1141 tcttctcgat ctcggaacag ctcaagaaat tccactcccg ggagttcccg aggaaccagc
     1201 ccagcaagga tggctggcaa tggtggagat gccgctctgg ccctacttct gctggaccgc
     1261 ctcaatcagt tagaatcaaa gatgtcgggg aaggggcagc aacagcaggg acagaccgta
     1321 actaaaaaga gtgcagcgga ggcctctaaa aagcctcggc agaaacgtac ggcaacaaag
     1381 gcttacaacg tgacccaggc ctttggccgt agaggacccg aacagacaca agggaacttt
     1441 ggtgaccaag agttgataag gcagggaacc gactacaaac actggcctca gatcgcccaa
     1501 ttcgcaccga gcgcgagcgc atttttcgga atgtcacgaa ttggcatgga ggtgacacct
     1561 tcgggaacct ggctgacata cactggcgcc ataaagctcg atgacaaaga tcccaacttc
     1621 aaggatcaag tcatcttgct gaataagcat attgacgcgt ataaaacttt tccacccacg
     1681 gaaccgaaga aggacaagaa aaagaaagct gatgagacac aggcgcttcc acaaaggcag
     1741 aaaaagcagc agacggttac tctactgccc gccgcagacc tagatgactt ctccaaacag
     1801 cttcagcagt ctatgagtag cgccgactca acccaagctg aaaaccttta ctttcagtcg
     1861 atggattata aggaccacga tggagactac aaagatcatg acattgatta caaggacgac
     1921 gatgacaaat aagctagctt gactgactga gatacagcgt accttcagct cacagacatg
     1981 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     2041 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     2101 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     2161 ttttaaagca agtaaaacct ctacaaatgt ggtattggcc catctctatc ggtatcgtag
     2221 cataacccct tggggcctct aaacgggtct tgaggggttt tttgtgcccc tcgggccgga
     2281 ttgctatcta ccggcattgg cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata
     2341 ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc acttccatac
     2401 gtgtcctcct taccagaaat ttatccttaa ggtcgtcagc tatcctgcag gcgatctctc
     2461 gatttcgatc aagacattcc tttaatggtc ttttctggac accactaggg gtcagaagta
     2521 gttcatcaaa ctttcttccc tccctaatct cattggttac cttgggctat cgaaacttaa
     2581 ttaaccagtc aagtcagcta cttggcgaga tcgacttgtc tgggtttcga ctacgctcag
     2641 aattgcgtca gtcaagttcg atctggtcct tgctattgca cccgttctcc gattacgagt
     2701 ttcatttaaa tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     2761 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     2821 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     2881 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     2941 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     3001 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     3061 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     3121 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     3181 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     3241 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     3301 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     3361 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     3421 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     3481 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     3541 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcattt
     3601 aaatttccga actctccaag gccctcgtcg gaaaatcttc aaacctttcg tccgatccat
     3661 cttgcaggct acctctcgaa cgaactatcg caagtctctt ggccggcctt gcgccttggc
     3721 tattgcttgg cagcgcctat cgccaggtat tactccaatc ccgaatatcc gagatcggga
     3781 tcacccgaga gaagttcaac ctacatcctc aatcccgatc tatccgagat ccgaggaata
     3841 tcgaaatcgg ggcgcgcctg gtgtaccgag aacgatcctc tcagtgcgag tctcgacgat
     3901 ccatatcgtt gcttggcagt cagccagtcg gaatccagct tgggacccag gaagtccaat
     3961 cgtcagatat tgtactcaag cctggtcacg gcagcgtacc gatctgttta aacctagata
     4021 ttgatagtct gatcggtcaa cgtataatcg agtcctagct tttgcaaaca tctatcaaga
     4081 gacaggatca gcaggaggct ttcgcatgag tattcaacat ttccgtgtcg cccttattcc
     4141 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     4201 agatgctgaa gatcagttgg gtgcgcgagt gggttacatc gaactggatc tcaacagcgg
     4261 taagatcctt gagagttttc gccccgaaga acgctttcca atgatgagca cttttaaagt
     4321 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     4381 catacactat tctcagaatg acttggttga gtattcacca gtcacagaaa agcatcttac
     4441 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     4501 ggccaactta cttctgacaa cgattggagg accgaaggag ctaaccgctt ttttgcacaa
     4561 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     4621 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaaccttgc gtaaactatt
     4681 aactggcgaa ctacttactc tagcttcccg gcaacagttg atagactgga tggaggcgga
     4741 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     4801 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     4861 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     4921 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaaccga ttctaggtgc
     4981 attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc acatatgcca
     5041 gattcagcaa cggatacggc ttccccaact tgcccacttc catacgtgtc ctccttacca
     5101 gaaatttatc cttaagatcc cgaatcgttt aaactcgact ctggctctat cgaatctccg
     5161 tcgtttcgag cttacgcgaa cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg
     5221 tcagctatcg tcagcttacc tttttggcag cgatcgcggc tcccgacatc ttggaccatt
     5281 agctccacag gtatcttctt ccctctagtg gtcataacag cagcttcagc tacctctcaa
     5341 ttcaaaaaac ccctcaagac ccgtttagag gccccaaggg gttatgctat caatcgttgc
     5401 gttacacaca caaaaaacca acacacatcc atcttcgatg gatagcgatt ttattatcta
     5461 actgctgatc gagtgtagcc agatctagta atcaattacg gggtcattag ttcatagccc
     5521 atatatggag ttccg
//
