LOCUS       sequence-295791-        7756 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 7756)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7756)
  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-m4133472
FEATURES             Location/Qualifiers
     source          1..7756
                     /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             1631..1648
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1725..1846
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1736..1755)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1790..1809
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1961..2007
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2037..2173
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2218..2289
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2515..3103)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2595..2614)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3603..4812
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4649..4668
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4816..5415
                     /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(4816..4835)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5312..5332
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5442..5490
                     /note="synthetic polyadenylation signal"
     CDS             join(5756..5824,5825..6616)
                     /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(5974..5993)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6629..6765
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6910..6981
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6986..7032)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7050..7098
                     /note="synthetic polyadenylation signal"
     enhancer        7179..7482
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7484..7686
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7637..7657
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgtct gataatggac cccaaaatca gcgaaatgca ccccgcatta cgtttggtgg
       61 accctcagat tcaactggca gtaaccagaa tggagaacgc agtggggcgc gatcaaaaca
      121 acgtcggccc caaggtttac ccaataatac tgcgtcttgg ttcaccgctc tcactcaaca
      181 tggcaaggaa gaccttaaat tccctcgagg acaaggcgtt ccaattaaca ccaatagcag
      241 tccagatgac caaattggct actaccgaag agctaccaga cgaattcgtg gtggtgacgg
      301 taaaatgaaa gatctcagtc caagatggta tttctactac ctaggaactg ggccagaagc
      361 tggacttccc tatggtgcta acaaagacgg catcatatgg gttgcaactg agggagcctt
      421 gaatacacca aaagatcaca ttggcacccg caatcctgct aacaatgctg caatcgtgct
      481 acaacttcct caaggaacaa cattgccaaa aggcttctac gcagaaggga gcagaggcgg
      541 cagtcaagcc tcttctcgtt cctcatcacg tagtcgcaac agttcaagaa attcaactcc
      601 aggcagcagt aggggaactt ctcctgctag aatggctggc aatggcggtg atgctgctct
      661 tgctttgctg ctgcttgaca gattgaacca gcttgagagc aaaatgtctg gtaaaggcca
      721 acaacaacaa ggccaaactg tcactaagaa atctgctgct gaggcttcta agaagcctcg
      781 gcaaaaacgt actgccacta aagcatacaa tgtaacacaa gctttcggca gacgtggtcc
      841 agaacaaacc caaggaaatt ttggggacca ggaactaatc agacaaggaa ctgattacaa
      901 acattggccg caaattgcac aatttgcccc cagcgcttca gcgttcttcg gaatgtcgcg
      961 cattggcatg gaagtcacac cttcgggaac gtggttgacc tacacaggtg ccatcaaatt
     1021 ggatgacaaa gatccaaatt tcaaagatca agtcattttg ctgaataagc atattgacgc
     1081 atacaaaaca ttcccaccaa cagagcctaa aaaggacaaa aagaagaagg ctgatgaaac
     1141 tcaagcctta ccgcagagac agaagaaaca gcaaactgtg actcttcttc ctgctgcaga
     1201 tttggatgat ttctccaaac aattgcaaca atccatgagc agtgctgact caactcaggc
     1261 cgaaaacctt tactttcagt cgatgagtga caaaatcatt catctaaccg atgactcctt
     1321 tgatacagac gtgttgaagg ccgatggtgc gatactggta gatttctggg ctgaatggtg
     1381 tggcccgtgc aagatgatcg cacccattct ggacgagatc gccgacgagt atcaaggaaa
     1441 attaactgtc gctaagctta atatagatca gaacccaggg acggcaccta aatacggcat
     1501 caggggaata cctaccctcc tgctctttaa gaatggggaa gttgccgcga caaaagtggg
     1561 agccctgtct aagggccagc taaaagagtt cttggacgct aacctggcag gttcaggctc
     1621 ggggcacatg caccatcacc atcaccatag cagcgggctc gtgccacgcg gctaagctag
     1681 cttgactgac tgagatacag cgtaccttca gctcacagac atgataagat acattgatga
     1741 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     1801 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     1861 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     1921 cctctacaaa tgtggtattg gcccatctct atcggtatcg tagcataacc ccttggggcc
     1981 tctaaacggg tcttgagggg ttttttgtgc ccctcgggcc ggattgctat ctaccggcat
     2041 tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga
     2101 ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga
     2161 aatttatcct taaggtcgtc agctatcctg caggcgatct ctcgatttcg atcaagacat
     2221 tcctttaatg gtcttttctg gacaccacta ggggtcagaa gtagttcatc aaactttctt
     2281 ccctccctaa tctcattggt taccttgggc tatcgaaact taattaacca gtcaagtcag
     2341 ctacttggcg agatcgactt gtctgggttt cgactacgct cagaattgcg tcagtcaagt
     2401 tcgatctggt ccttgctatt gcacccgttc tccgattacg agtttcattt aaatcatgtg
     2461 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2521 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2581 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     2641 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     2701 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     2761 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     2821 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     2881 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     2941 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     3001 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     3061 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     3121 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     3181 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     3241 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     3301 tatctcagcg atctgtctat ttcgttcatc catagttgca tttaaatttc cgaactctcc
     3361 aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc catcttgcag gctacctctc
     3421 gaacgaacta tcgcaagtct cttggccggc cttgcgcctt ggctattgct tggcagcgcc
     3481 tatcgccagg tattactcca atcccgaata tccgagatcg ggatcacccg agagaagttc
     3541 aacctacatc ctcaatcccg atctatccga gatccgagga atatcgaaat cggggcgcgc
     3601 ctggcctccg cgccgggttt tggcgcctcc cgcgggcgcc cccctcgtca cggcgagcgc
     3661 tgccacgtca gacgaagggc gcaggagcgt cctgatcctt ccgcccggac gctcaggaca
     3721 gcggcccgct gctcataaga ctcggcctta gaaccccagt atcagcagaa ggacatttta
     3781 ggacgggact tgggtgactc tagggcactg gttttctttc cagagagcgg aacaggcgag
     3841 gaaaagtagt cccttctcgg cgattctgcg gagggatctc cgtggggcgg tgaacgccga
     3901 tgattatata aggacgcgcc gggtgtggca cagctagttc cgtcgcagcc gggatttggg
     3961 tcgcggttct tgtttgtgga tcgctgtgat cgtcacttgg tgagtagcgg gctgctgggc
     4021 tggccggggc tttcgtggcc gccgggccgc tcggtgggac ggaagcgtgt ggagagaccg
     4081 ccaagggctg tagtctgggt ccgcgagcaa ggttgccctg aactgggggt tggggggagc
     4141 gcagcaaaat ggcggctgtt cccgagtctt gaatggaaga cgcttgtgag gcgggctgtg
     4201 aggtcgttga aacaaggtgg ggggcatggt gggcggcaag aacccaaggt cttgagccct
     4261 tcgctaatgc gggaaagctc ttattcgggt gagatgggct gggcaccatc tggggaccct
     4321 gacgtgaagt ttgtcactga ctggagaact cggtttgtcg tctgttgcgg gggcggcagt
     4381 tatggcggtg ccgttgggca gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc
     4441 gtgacgtcac ccgttctgtt ggcttataat gcagggtggg gccacctgcc ggtaggtgtg
     4501 cggtaggctt ttctccgtcg caggacgcag ggttcgggcc tagggtaggc tctcctgaat
     4561 cgacaggcgc cggacctctg gtgaggggag ggataagtga ggcgtcagtt tctttggtcg
     4621 gttttatgta cctatcttct taagtagctg aagctccggt tttgaactat gcgctcgggg
     4681 ttggcgagtg tgttttgtga agttttttag gcaccttttg aaatgtaatc atttgggtca
     4741 atatgtaatt ttcagtgtta gacttgtaaa ttgtccgcta aattctggcc gtttttggct
     4801 tttttgttag acaacatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac
     4861 gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac
     4921 accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg
     4981 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc
     5041 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg
     5101 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggagggcct cctggcgccg
     5161 caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag
     5221 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggccgccga gcgcgccggg
     5281 gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc
     5341 ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc
     5401 aagcccggtg cctaggctag tatgtaagcc tagtcttaga taataaaatc gctatccatc
     5461 gaagatggat gtgtgttggt tttttgtgtg tgtaacgcta ggcgcgcctg gtgtaccgag
     5521 aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt gcttggcagt cagccagtcg
     5581 gaatccagct tgggacccag gaagtccaat cgtcagatat tgtactcaag cctggtcacg
     5641 gcagcgtacc gatctgttta aacctagata ttgatagtct gatcggtcaa cgtataatcg
     5701 agtcctagct tttgcaaaca tctatcaaga gacaggatca gcaggaggct ttcgcatgag
     5761 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     5821 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcgcgagt
     5881 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     5941 acgctttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     6001 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     6061 gtattcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     6121 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgattggagg
     6181 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     6241 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     6301 agcaatggca acaaccttgc gtaaactatt aactggcgaa ctacttactc tagcttcccg
     6361 gcaacagttg atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     6421 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     6481 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     6541 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     6601 gattaagcat tggtaaccga ttctaggtgc attggcgcag aaaaaaatgc ctgatgcgac
     6661 gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc ttccccaact
     6721 tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaagatcg tttaaactcg
     6781 actctggctc tatcgaatct ccgtcgtttc gagcttacgc gaacagccgt ggcgctcatt
     6841 tgctcgtcgg gcatcgaatc tcgtcagcta tcgtcagctt acctttttgg cagcgatcgc
     6901 ggctcccgac atcttggacc attagctcca caggtatctt cttccctcta gtggtcataa
     6961 cagcagcttc agctacctct caattcaaaa aacccctcaa gacccgttta gaggccccaa
     7021 ggggttatgc tatcaatcgt tgcgttacac acacaaaaaa ccaacacaca tccatcttcg
     7081 atggatagcg attttattat ctaactgctg atcgagtgta gccagatcta gtaatcaatt
     7141 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     7201 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     7261 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     7321 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     7381 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     7441 acttggcagt acatctacgt attagtcatc gctattacca tgctgatgcg gttttggcag
     7501 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
     7561 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
     7621 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
     7681 agagctggtt tagtgaaccg tcagatcaga tctttgtcga tcctaccatc cactcgacac
     7741 acccgccagc ggccgc
//
