LOCUS       sequence-300259-        7352 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 7352)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7352)
  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-m4134903
FEATURES             Location/Qualifiers
     source          1..7352
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1314..1435
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1325..1344)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1379..1398
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1550..1596
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1626..1762
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1807..1878
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2104..2692)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2184..2203)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3192..4401
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4238..4257
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4405..5004
                     /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(4405..4424)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4901..4921
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5031..5079
                     /note="synthetic polyadenylation signal"
     CDS             join(5345..5413,5414..6205)
                     /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(5563..5582)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6218..6354
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6506..6577
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6582..6628)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6646..6694
                     /note="synthetic polyadenylation signal"
     enhancer        6775..7078
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7080..7282
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7233..7253
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgtcg gataacggtc cgcagaatca acggaatgct ccaaggatta cgttcggagg
       61 cccctcagac agtactggga gcaaccagaa tggtgaaaga tctggcgcgc gctccaaaca
      121 gcgaagacct caaggattac ccaacaacac agcatcctgg tttaccgccc taactcagca
      181 cgggaaggag gatcttaagt ttcctcgtgg ccagggggtg ccaatcaata ccaacagttc
      241 ccctgacgac cagataggat attacaggcg ggccacccgc cggatccgag gcggagatgg
      301 taaaatgaag gatttgagcc cccgctggta cttctattac ctcggaacag gcccagaggc
      361 tgggctgcct tatggagcaa ataaagacgg cattatatgg gtagccacgg aaggggcact
      421 gaacacccca aaggaccata tcggcactag aaatcccgcg aacaatgccg ctattgttct
      481 gcaacttccc cagggaacaa ccctgccgaa aggtttctat gctgagggct caaggggcgg
      541 gtcgcaggcc tctagtagaa gctcttcacg gagcagaaac tccagcagga acagtactcc
      601 ggggtcttcc cgcggaacat cccctgcccg tatggcgggt aacgggggtg atgccgcact
      661 agctcttctg ctcttggaca gactgaatca actcgaatcg aagatgtcag gcaaaggaca
      721 gcagcaacag ggacagaccg tgaccaagaa gagtgccgca gaggctagca agaaaccccg
      781 acaaaaacgc acggccacaa aggcgtacaa tgtcactcag gcttttggtc gcagaggccc
      841 cgagcaaaca cagggcaatt ttggcgacca ggaactgatc aggcagggaa ccgattacaa
      901 acactggcct cagattgccc agttcgcccc atcggcatct gcttttttcg ggatgtcacg
      961 gatcggcatg gaggtgactc cgtcaggtac gtggctgacc tacacaggcg caataaaatt
     1021 agatgacaag gatccaaact tcaaggatca agttattcta ctgaataagc atatcgacgc
     1081 atataaaact tttcccccta ctgaacccaa gaaagataaa aaaaagaaag ctgacgaaac
     1141 tcaagcactc ccacagcggc agaagaagca gcagacagtc acattgttgc cagccgcgga
     1201 cctcgatgat ttctctaagc aactccagca gagcatgtcc agtgccgact ccacccaagc
     1261 ttaagctagc ttgactgact gagatacagc gtaccttcag ctcacagaca tgataagata
     1321 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     1381 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     1441 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag
     1501 caagtaaaac ctctacaaat gtggtattgg cccatctcta tcggtatcgt agcataaccc
     1561 cttggggcct ctaaacgggt cttgaggggt tttttgtgcc cctcgggccg gattgctatc
     1621 taccggcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca
     1681 tatgccagat tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc
     1741 cttaccagaa atttatcctt aaggtcgtca gctatcctgc aggcgatctc tcgatttcga
     1801 tcaagacatt cctttaatgg tcttttctgg acaccactag gggtcagaag tagttcatca
     1861 aactttcttc cctccctaat ctcattggtt accttgggct atcgaaactt aattaaccag
     1921 tcaagtcagc tacttggcga gatcgacttg tctgggtttc gactacgctc agaattgcgt
     1981 cagtcaagtt cgatctggtc cttgctattg cacccgttct ccgattacga gtttcattta
     2041 aatcatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     2101 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     2161 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     2221 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2281 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2341 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2401 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     2461 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     2521 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     2581 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2641 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     2701 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     2761 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     2821 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     2881 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcat ttaaatttcc
     2941 gaactctcca aggccctcgt cggaaaatct tcaaaccttt cgtccgatcc atcttgcagg
     3001 ctacctctcg aacgaactat cgcaagtctc ttggccggcc ttgcgccttg gctattgctt
     3061 ggcagcgcct atcgccaggt attactccaa tcccgaatat ccgagatcgg gatcacccga
     3121 gagaagttca acctacatcc tcaatcccga tctatccgag atccgaggaa tatcgaaatc
     3181 ggggcgcgcc tggcctccgc gccgggtttt ggcgcctccc gcgggcgccc ccctcgtcac
     3241 ggcgagcgct gccacgtcag acgaagggcg caggagcgtc ctgatccttc cgcccggacg
     3301 ctcaggacag cggcccgctg ctcataagac tcggccttag aaccccagta tcagcagaag
     3361 gacattttag gacgggactt gggtgactct agggcactgg ttttctttcc agagagcgga
     3421 acaggcgagg aaaagtagtc ccttctcggc gattctgcgg agggatctcc gtggggcggt
     3481 gaacgccgat gattatataa ggacgcgccg ggtgtggcac agctagttcc gtcgcagccg
     3541 ggatttgggt cgcggttctt gtttgtggat cgctgtgatc gtcacttggt gagtagcggg
     3601 ctgctgggct ggccggggct ttcgtggccg ccgggccgct cggtgggacg gaagcgtgtg
     3661 gagagaccgc caagggctgt agtctgggtc cgcgagcaag gttgccctga actgggggtt
     3721 ggggggagcg cagcaaaatg gcggctgttc ccgagtcttg aatggaagac gcttgtgagg
     3781 cgggctgtga ggtcgttgaa acaaggtggg gggcatggtg ggcggcaaga acccaaggtc
     3841 ttgagccctt cgctaatgcg ggaaagctct tattcgggtg agatgggctg ggcaccatct
     3901 ggggaccctg acgtgaagtt tgtcactgac tggagaactc ggtttgtcgt ctgttgcggg
     3961 ggcggcagtt atggcggtgc cgttgggcag tgcacccgta cctttgggag cgcgcgccct
     4021 cgtcgtgtcg tgacgtcacc cgttctgttg gcttataatg cagggtgggg ccacctgccg
     4081 gtaggtgtgc ggtaggcttt tctccgtcgc aggacgcagg gttcgggcct agggtaggct
     4141 ctcctgaatc gacaggcgcc ggacctctgg tgaggggagg gataagtgag gcgtcagttt
     4201 ctttggtcgg ttttatgtac ctatcttctt aagtagctga agctccggtt ttgaactatg
     4261 cgctcggggt tggcgagtgt gttttgtgaa gttttttagg caccttttga aatgtaatca
     4321 tttgggtcaa tatgtaattt tcagtgttag acttgtaaat tgtccgctaa attctggccg
     4381 tttttggctt ttttgttaga caacatgacc gagtacaagc ccacggtgcg cctcgccacc
     4441 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc
     4501 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc
     4561 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg
     4621 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc
     4681 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggagggcctc
     4741 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc
     4801 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggccgccgag
     4861 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag
     4921 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc
     4981 atgacccgca agcccggtgc ctaggctagt atgtaagcct agtcttagat aataaaatcg
     5041 ctatccatcg aagatggatg tgtgttggtt ttttgtgtgt gtaacgctag gcgcgcctgg
     5101 tgtaccgaga acgatcctct cagtgcgagt ctcgacgatc catatcgttg cttggcagtc
     5161 agccagtcgg aatccagctt gggacccagg aagtccaatc gtcagatatt gtactcaagc
     5221 ctggtcacgg cagcgtaccg atctgtttaa acctagatat tgatagtctg atcggtcaac
     5281 gtataatcga gtcctagctt ttgcaaacat ctatcaagag acaggatcag caggaggctt
     5341 tcgcatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     5401 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     5461 tgcgcgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     5521 ccccgaagaa cgctttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     5581 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     5641 cttggttgag tattcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     5701 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     5761 gattggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     5821 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     5881 gatgcctgta gcaatggcaa caaccttgcg taaactatta actggcgaac tacttactct
     5941 agcttcccgg caacagttga tagactggat ggaggcggat aaagttgcag gaccacttct
     6001 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     6061 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     6121 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     6181 tgcctcactg attaagcatt ggtaaccgat tctaggtgca ttggcgcaga aaaaaatgcc
     6241 tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac ggatacggct
     6301 tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc ttaagatccc
     6361 gaatcgttta aactcgactc tggctctatc gaatctccgt cgtttcgagc ttacgcgaac
     6421 agccgtggcg ctcatttgct cgtcgggcat cgaatctcgt cagctatcgt cagcttacct
     6481 ttttggcagc gatcgcggct cccgacatct tggaccatta gctccacagg tatcttcttc
     6541 cctctagtgg tcataacagc agcttcagct acctctcaat tcaaaaaacc cctcaagacc
     6601 cgtttagagg ccccaagggg ttatgctatc aatcgttgcg ttacacacac aaaaaaccaa
     6661 cacacatcca tcttcgatgg atagcgattt tattatctaa ctgctgatcg agtgtagcca
     6721 gatctagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
     6781 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
     6841 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
     6901 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
     6961 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
     7021 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatgct
     7081 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
     7141 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     7201 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
     7261 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atcagatctt tgtcgatcct
     7321 accatccact cgacacaccc gccagcggcc gc
//
