LOCUS       sequence-307265-        7763 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 7763)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7763)
  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-m4134554
FEATURES             Location/Qualifiers
     source          1..7763
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        14..317
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        319..521
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     472..492
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1853..1873
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2222..2239
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2316..2437
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2327..2346)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2381..2400
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2552..2598
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2628..2764
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2809..2880
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3106..3694)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3186..3205)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4194..5403
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5240..5259
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5407..6006
                     /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(5407..5426)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5903..5923
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    6033..6081
                     /note="synthetic polyadenylation signal"
     CDS             join(6347..6415,6416..7207)
                     /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(6565..6584)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7220..7356
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7508..7579
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7584..7630)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7648..7696
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
       61 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      121 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      181 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      241 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      301 tcatcgctat taccatgctg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      361 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      421 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      481 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga
      541 tcagatcttt gtcgatccta ccatccactc gacacacccg ccagcggccg ccaccatgtc
      601 tgataatgga ccacagaacc aacgcaatgc accccgaatc acttttggcg gtccgtccga
      661 cagtaccggg agcaaccaga acggcgaaag atcgggagct cgttcaaaac agcggaggcc
      721 tcaagggtta cccaataaca cggccagctg gttcacagcg ttgacccagc atggcaagga
      781 ggacctcaag ttccctcggg gtcagggggt cccaataaat acaaacagtt cccctgatga
      841 ccaaattgga tactatagac gcgctactag gaggatccgt ggcggggatg gaaaaatgaa
      901 ggacctttct cccagatggt acttttatta cctgggtact ggcccagagg ccggtctacc
      961 gtatggagca aacaaagatg gcattatttg ggtggccacg gaaggagcac tgaatacccc
     1021 aaaggatcac atcggcaccc ggaatccagc caacaatgct gctatcgttc tgcaactccc
     1081 ccaggggaca acattgccta aaggcttcta tgccgagggc agtagaggtg gcagccaggc
     1141 gtcctcacgc agctcttctc gatctcggaa cagctcaaga aattccactc ccgggagttc
     1201 ccgaggaacc agcccagcaa ggatggctgg caatggtgga gatgccgctc tggccctact
     1261 tctgctggac cgcctcaatc agttagaatc aaagatgtcg gggaaggggc agcaacagca
     1321 gggacagacc gtaactaaaa agagtgcagc ggaggcctct aaaaagcctc ggcagaaacg
     1381 tacggcaaca aaggcttaca acgtgaccca ggcctttggc cgtagaggac ccgaacagac
     1441 acaagggaac tttggtgacc aagagttgat aaggcaggga accgactaca aacactggcc
     1501 tcagatcgcc caattcgcac cgagcgcgag cgcatttttc ggaatgtcac gaattggcat
     1561 ggaggtgaca ccttcgggaa cctggctgac atacactggc gccataaagc tcgatgacaa
     1621 agatcccaac ttcaaggatc aagtcatctt gctgaataag catattgacg cgtataaaac
     1681 ttttccaccc acggaaccga agaaggacaa gaaaaagaaa gctgatgaga cacaggcgct
     1741 tccacaaagg cagaaaaagc agcagacggt tactctactg cccgccgcag acctagatga
     1801 cttctccaaa cagcttcagc agtctatgag tagcgccgac tcaacccaag ctgaaaacct
     1861 ttactttcag tcgatgagtg acaaaatcat tcatctaacc gatgactcct ttgatacaga
     1921 cgtgttgaag gccgatggtg cgatactggt agatttctgg gctgaatggt gtggcccgtg
     1981 caagatgatc gcacccattc tggacgagat cgccgacgag tatcaaggaa aattaactgt
     2041 cgctaagctt aatatagatc agaacccagg gacggcacct aaatacggca tcaggggaat
     2101 acctaccctc ctgctcttta agaatgggga agttgccgcg acaaaagtgg gagccctgtc
     2161 taagggccag ctaaaagagt tcttggacgc taacctggca ggttcaggct cggggcacat
     2221 gcaccatcac catcaccata gcagcgggct cgtgccacgc ggctaagcta gcttgactga
     2281 ctgagataca gcgtaccttc agctcacaga catgataaga tacattgatg agtttggaca
     2341 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     2401 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     2461 tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa
     2521 atgtggtatt ggcccatctc tatcggtatc gtagcataac cccttggggc ctctaaacgg
     2581 gtcttgaggg gttttttgtg cccctcgggc cggattgcta tctaccggca ttggcgcaga
     2641 aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac
     2701 ggatacggct tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc
     2761 ttaaggtcgt cagctatcct gcaggcgatc tctcgatttc gatcaagaca ttcctttaat
     2821 ggtcttttct ggacaccact aggggtcaga agtagttcat caaactttct tccctcccta
     2881 atctcattgg ttaccttggg ctatcgaaac ttaattaacc agtcaagtca gctacttggc
     2941 gagatcgact tgtctgggtt tcgactacgc tcagaattgc gtcagtcaag ttcgatctgg
     3001 tccttgctat tgcacccgtt ctccgattac gagtttcatt taaatcatgt gagcaaaagg
     3061 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     3121 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3181 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3241 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3301 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3361 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3421 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3481 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     3541 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     3601 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     3661 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     3721 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     3781 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     3841 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     3901 gatctgtcta tttcgttcat ccatagttgc atttaaattt ccgaactctc caaggccctc
     3961 gtcggaaaat cttcaaacct ttcgtccgat ccatcttgca ggctacctct cgaacgaact
     4021 atcgcaagtc tcttggccgg ccttgcgcct tggctattgc ttggcagcgc ctatcgccag
     4081 gtattactcc aatcccgaat atccgagatc gggatcaccc gagagaagtt caacctacat
     4141 cctcaatccc gatctatccg agatccgagg aatatcgaaa tcggggcgcg cctggcctcc
     4201 gcgccgggtt ttggcgcctc ccgcgggcgc ccccctcgtc acggcgagcg ctgccacgtc
     4261 agacgaaggg cgcaggagcg tcctgatcct tccgcccgga cgctcaggac agcggcccgc
     4321 tgctcataag actcggcctt agaaccccag tatcagcaga aggacatttt aggacgggac
     4381 ttgggtgact ctagggcact ggttttcttt ccagagagcg gaacaggcga ggaaaagtag
     4441 tcccttctcg gcgattctgc ggagggatct ccgtggggcg gtgaacgccg atgattatat
     4501 aaggacgcgc cgggtgtggc acagctagtt ccgtcgcagc cgggatttgg gtcgcggttc
     4561 ttgtttgtgg atcgctgtga tcgtcacttg gtgagtagcg ggctgctggg ctggccgggg
     4621 ctttcgtggc cgccgggccg ctcggtggga cggaagcgtg tggagagacc gccaagggct
     4681 gtagtctggg tccgcgagca aggttgccct gaactggggg ttggggggag cgcagcaaaa
     4741 tggcggctgt tcccgagtct tgaatggaag acgcttgtga ggcgggctgt gaggtcgttg
     4801 aaacaaggtg gggggcatgg tgggcggcaa gaacccaagg tcttgagccc ttcgctaatg
     4861 cgggaaagct cttattcggg tgagatgggc tgggcaccat ctggggaccc tgacgtgaag
     4921 tttgtcactg actggagaac tcggtttgtc gtctgttgcg ggggcggcag ttatggcggt
     4981 gccgttgggc agtgcacccg tacctttggg agcgcgcgcc ctcgtcgtgt cgtgacgtca
     5041 cccgttctgt tggcttataa tgcagggtgg ggccacctgc cggtaggtgt gcggtaggct
     5101 tttctccgtc gcaggacgca gggttcgggc ctagggtagg ctctcctgaa tcgacaggcg
     5161 ccggacctct ggtgagggga gggataagtg aggcgtcagt ttctttggtc ggttttatgt
     5221 acctatcttc ttaagtagct gaagctccgg ttttgaacta tgcgctcggg gttggcgagt
     5281 gtgttttgtg aagtttttta ggcacctttt gaaatgtaat catttgggtc aatatgtaat
     5341 tttcagtgtt agacttgtaa attgtccgct aaattctggc cgtttttggc ttttttgtta
     5401 gacaacatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg
     5461 gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat
     5521 ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg
     5581 ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg
     5641 ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg
     5701 agcggttccc ggctggccgc gcagcaacag atggagggcc tcctggcgcc gcaccggccc
     5761 aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt
     5821 ctgggcagcg ccgtcgtgct ccccggagtg gaggccgccg agcgcgccgg ggtgcccgcc
     5881 ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc
     5941 accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt
     6001 gcctaggcta gtatgtaagc ctagtcttag ataataaaat cgctatccat cgaagatgga
     6061 tgtgtgttgg ttttttgtgt gtgtaacgct aggcgcgcct ggtgtaccga gaacgatcct
     6121 ctcagtgcga gtctcgacga tccatatcgt tgcttggcag tcagccagtc ggaatccagc
     6181 ttgggaccca ggaagtccaa tcgtcagata ttgtactcaa gcctggtcac ggcagcgtac
     6241 cgatctgttt aaacctagat attgatagtc tgatcggtca acgtataatc gagtcctagc
     6301 ttttgcaaac atctatcaag agacaggatc agcaggaggc tttcgcatga gtattcaaca
     6361 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     6421 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcgcgag tgggttacat
     6481 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgctttcc
     6541 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     6601 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtattcacc
     6661 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     6721 aaccatgagt gataacactg cggccaactt acttctgaca acgattggag gaccgaagga
     6781 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     6841 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     6901 aacaaccttg cgtaaactat taactggcga actacttact ctagcttccc ggcaacagtt
     6961 gatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     7021 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     7081 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     7141 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     7201 ttggtaaccg attctaggtg cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt
     7261 cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac ttgcccactt
     7321 ccatacgtgt cctccttacc agaaatttat ccttaagatc ccgaatcgtt taaactcgac
     7381 tctggctcta tcgaatctcc gtcgtttcga gcttacgcga acagccgtgg cgctcatttg
     7441 ctcgtcgggc atcgaatctc gtcagctatc gtcagcttac ctttttggca gcgatcgcgg
     7501 ctcccgacat cttggaccat tagctccaca ggtatcttct tccctctagt ggtcataaca
     7561 gcagcttcag ctacctctca attcaaaaaa cccctcaaga cccgtttaga ggccccaagg
     7621 ggttatgcta tcaatcgttg cgttacacac acaaaaaacc aacacacatc catcttcgat
     7681 ggatagcgat tttattatct aactgctgat cgagtgtagc cagatctagt aatcaattac
     7741 ggggtcatta gttcatagcc cat
//
