LOCUS       sequence-320770-        7442 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 7442)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7442)
  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-m4134552
FEATURES             Location/Qualifiers
     source          1..7442
                     /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"
     promoter        3860..5069
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4906..4925
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5073..5672
                     /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(5073..5092)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5569..5589
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5699..5747
                     /note="synthetic polyadenylation signal"
     CDS             join(6013..6081,6082..6873)
                     /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(6231..6250)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6886..7022
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7174..7245
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7250..7296)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7314..7362
                     /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 gcctccgcgc cgggttttgg cgcctcccgc gggcgccccc
     3901 ctcgtcacgg cgagcgctgc cacgtcagac gaagggcgca ggagcgtcct gatccttccg
     3961 cccggacgct caggacagcg gcccgctgct cataagactc ggccttagaa ccccagtatc
     4021 agcagaagga cattttagga cgggacttgg gtgactctag ggcactggtt ttctttccag
     4081 agagcggaac aggcgaggaa aagtagtccc ttctcggcga ttctgcggag ggatctccgt
     4141 ggggcggtga acgccgatga ttatataagg acgcgccggg tgtggcacag ctagttccgt
     4201 cgcagccggg atttgggtcg cggttcttgt ttgtggatcg ctgtgatcgt cacttggtga
     4261 gtagcgggct gctgggctgg ccggggcttt cgtggccgcc gggccgctcg gtgggacgga
     4321 agcgtgtgga gagaccgcca agggctgtag tctgggtccg cgagcaaggt tgccctgaac
     4381 tgggggttgg ggggagcgca gcaaaatggc ggctgttccc gagtcttgaa tggaagacgc
     4441 ttgtgaggcg ggctgtgagg tcgttgaaac aaggtggggg gcatggtggg cggcaagaac
     4501 ccaaggtctt gagcccttcg ctaatgcggg aaagctctta ttcgggtgag atgggctggg
     4561 caccatctgg ggaccctgac gtgaagtttg tcactgactg gagaactcgg tttgtcgtct
     4621 gttgcggggg cggcagttat ggcggtgccg ttgggcagtg cacccgtacc tttgggagcg
     4681 cgcgccctcg tcgtgtcgtg acgtcacccg ttctgttggc ttataatgca gggtggggcc
     4741 acctgccggt aggtgtgcgg taggcttttc tccgtcgcag gacgcagggt tcgggcctag
     4801 ggtaggctct cctgaatcga caggcgccgg acctctggtg aggggaggga taagtgaggc
     4861 gtcagtttct ttggtcggtt ttatgtacct atcttcttaa gtagctgaag ctccggtttt
     4921 gaactatgcg ctcggggttg gcgagtgtgt tttgtgaagt tttttaggca ccttttgaaa
     4981 tgtaatcatt tgggtcaata tgtaattttc agtgttagac ttgtaaattg tccgctaaat
     5041 tctggccgtt tttggctttt ttgttagaca acatgaccga gtacaagccc acggtgcgcc
     5101 tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact
     5161 accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc
     5221 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg
     5281 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg
     5341 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg
     5401 agggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg
     5461 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg
     5521 ccgccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct
     5581 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca
     5641 cctggtgcat gacccgcaag cccggtgcct aggctagtat gtaagcctag tcttagataa
     5701 taaaatcgct atccatcgaa gatggatgtg tgttggtttt ttgtgtgtgt aacgctaggc
     5761 gcgcctggtg taccgagaac gatcctctca gtgcgagtct cgacgatcca tatcgttgct
     5821 tggcagtcag ccagtcggaa tccagcttgg gacccaggaa gtccaatcgt cagatattgt
     5881 actcaagcct ggtcacggca gcgtaccgat ctgtttaaac ctagatattg atagtctgat
     5941 cggtcaacgt ataatcgagt cctagctttt gcaaacatct atcaagagac aggatcagca
     6001 ggaggctttc gcatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     6061 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     6121 cagttgggtg cgcgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     6181 agttttcgcc ccgaagaacg ctttccaatg atgagcactt ttaaagttct gctatgtggc
     6241 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     6301 cagaatgact tggttgagta ttcaccagtc acagaaaagc atcttacgga tggcatgaca
     6361 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     6421 ctgacaacga ttggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     6481 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     6541 gacaccacga tgcctgtagc aatggcaaca accttgcgta aactattaac tggcgaacta
     6601 cttactctag cttcccggca acagttgata gactggatgg aggcggataa agttgcagga
     6661 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     6721 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     6781 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     6841 gagataggtg cctcactgat taagcattgg taaccgattc taggtgcatt ggcgcagaaa
     6901 aaaatgcctg atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg
     6961 atacggcttc cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt
     7021 aagatcccga atcgtttaaa ctcgactctg gctctatcga atctccgtcg tttcgagctt
     7081 acgcgaacag ccgtggcgct catttgctcg tcgggcatcg aatctcgtca gctatcgtca
     7141 gcttaccttt ttggcagcga tcgcggctcc cgacatcttg gaccattagc tccacaggta
     7201 tcttcttccc tctagtggtc ataacagcag cttcagctac ctctcaattc aaaaaacccc
     7261 tcaagacccg tttagaggcc ccaaggggtt atgctatcaa tcgttgcgtt acacacacaa
     7321 aaaaccaaca cacatccatc ttcgatggat agcgatttta ttatctaact gctgatcgag
     7381 tgtagccaga tctagtaatc aattacgggg tcattagttc atagcccata tatggagttc
     7441 cg
//
