LOCUS       sequence-306156-        7418 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 7418)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7418)
  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-m4134558
FEATURES             Location/Qualifiers
     source          1..7418
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    86..157
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(162..208)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    226..274
                     /note="synthetic polyadenylation signal"
     enhancer        355..658
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        660..862
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     813..833
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2194..2214
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2215..2259
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    2312..2433
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2323..2342)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2377..2396
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2548..2594
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2624..2760
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2805..2876
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3102..3690)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3182..3201)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4190..5399
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5236..5255
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5403..6002
                     /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(5403..5422)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5899..5919
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    6029..6077
                     /note="synthetic polyadenylation signal"
     CDS             join(6343..6411,6412..7203)
                     /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(6561..6580)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7216..7352
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 agccgtggcg ctcatttgct cgtcgggcat cgaatctcgt cagctatcgt cagcttacct
       61 ttttggcagc gatcgcggct cccgacatct tggaccatta gctccacagg tatcttcttc
      121 cctctagtgg tcataacagc agcttcagct acctctcaat tcaaaaaacc cctcaagacc
      181 cgtttagagg ccccaagggg ttatgctatc aatcgttgcg ttacacacac aaaaaaccaa
      241 cacacatcca tcttcgatgg atagcgattt tattatctaa ctgctgatcg agtgtagcca
      301 gatctagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      361 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      421 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      481 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      541 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      601 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatgct
      661 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      721 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      781 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      841 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atcagatctt tgtcgatcct
      901 accatccact cgacacaccc gccagcggcc gccaccatgt ctgataatgg accacagaac
      961 caacgcaatg caccccgaat cacttttggc ggtccgtccg acagtaccgg gagcaaccag
     1021 aacggcgaaa gatcgggagc tcgttcaaaa cagcggaggc ctcaagggtt acccaataac
     1081 acggccagct ggttcacagc gttgacccag catggcaagg aggacctcaa gttccctcgg
     1141 ggtcaggggg tcccaataaa tacaaacagt tcccctgatg accaaattgg atactataga
     1201 cgcgctacta ggaggatccg tggcggggat ggaaaaatga aggacctttc tcccagatgg
     1261 tacttttatt acctgggtac tggcccagag gccggtctac cgtatggagc aaacaaagat
     1321 ggcattattt gggtggccac ggaaggagca ctgaataccc caaaggatca catcggcacc
     1381 cggaatccag ccaacaatgc tgctatcgtt ctgcaactcc cccaggggac aacattgcct
     1441 aaaggcttct atgccgaggg cagtagaggt ggcagccagg cgtcctcacg cagctcttct
     1501 cgatctcgga acagctcaag aaattccact cccgggagtt cccgaggaac cagcccagca
     1561 aggatggctg gcaatggtgg agatgccgct ctggccctac ttctgctgga ccgcctcaat
     1621 cagttagaat caaagatgtc ggggaagggg cagcaacagc agggacagac cgtaactaaa
     1681 aagagtgcag cggaggcctc taaaaagcct cggcagaaac gtacggcaac aaaggcttac
     1741 aacgtgaccc aggcctttgg ccgtagagga cccgaacaga cacaagggaa ctttggtgac
     1801 caagagttga taaggcaggg aaccgactac aaacactggc ctcagatcgc ccaattcgca
     1861 ccgagcgcga gcgcattttt cggaatgtca cgaattggca tggaggtgac accttcggga
     1921 acctggctga catacactgg cgccataaag ctcgatgaca aagatcccaa cttcaaggat
     1981 caagtcatct tgctgaataa gcatattgac gcgtataaaa cttttccacc cacggaaccg
     2041 aagaaggaca agaaaaagaa agctgatgag acacaggcgc ttccacaaag gcagaaaaag
     2101 cagcagacgg ttactctact gcccgccgca gacctagatg acttctccaa acagcttcag
     2161 cagtctatga gtagcgccga ctcaacccaa gctgaaaacc tttactttca gtcgaaagaa
     2221 accgcggcag ctaagttcga gaggcagcac atggacagtt aagctagctt gactgactga
     2281 gatacagcgt accttcagct cacagacatg ataagataca ttgatgagtt tggacaaacc
     2341 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     2401 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     2461 tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt
     2521 ggtattggcc catctctatc ggtatcgtag cataacccct tggggcctct aaacgggtct
     2581 tgaggggttt tttgtgcccc tcgggccgga ttgctatcta ccggcattgg cgcagaaaaa
     2641 aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc agcaacggat
     2701 acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat ttatccttaa
     2761 ggtcgtcagc tatcctgcag gcgatctctc gatttcgatc aagacattcc tttaatggtc
     2821 ttttctggac accactaggg gtcagaagta gttcatcaaa ctttcttccc tccctaatct
     2881 cattggttac cttgggctat cgaaacttaa ttaaccagtc aagtcagcta cttggcgaga
     2941 tcgacttgtc tgggtttcga ctacgctcag aattgcgtca gtcaagttcg atctggtcct
     3001 tgctattgca cccgttctcc gattacgagt ttcatttaaa tcatgtgagc aaaaggccag
     3061 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     3121 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     3181 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     3241 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     3301 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     3361 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     3421 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     3481 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     3541 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     3601 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     3661 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     3721 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     3781 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     3841 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     3901 tgtctatttc gttcatccat agttgcattt aaatttccga actctccaag gccctcgtcg
     3961 gaaaatcttc aaacctttcg tccgatccat cttgcaggct acctctcgaa cgaactatcg
     4021 caagtctctt ggccggcctt gcgccttggc tattgcttgg cagcgcctat cgccaggtat
     4081 tactccaatc ccgaatatcc gagatcggga tcacccgaga gaagttcaac ctacatcctc
     4141 aatcccgatc tatccgagat ccgaggaata tcgaaatcgg ggcgcgcctg gcctccgcgc
     4201 cgggttttgg cgcctcccgc gggcgccccc ctcgtcacgg cgagcgctgc cacgtcagac
     4261 gaagggcgca ggagcgtcct gatccttccg cccggacgct caggacagcg gcccgctgct
     4321 cataagactc ggccttagaa ccccagtatc agcagaagga cattttagga cgggacttgg
     4381 gtgactctag ggcactggtt ttctttccag agagcggaac aggcgaggaa aagtagtccc
     4441 ttctcggcga ttctgcggag ggatctccgt ggggcggtga acgccgatga ttatataagg
     4501 acgcgccggg tgtggcacag ctagttccgt cgcagccggg atttgggtcg cggttcttgt
     4561 ttgtggatcg ctgtgatcgt cacttggtga gtagcgggct gctgggctgg ccggggcttt
     4621 cgtggccgcc gggccgctcg gtgggacgga agcgtgtgga gagaccgcca agggctgtag
     4681 tctgggtccg cgagcaaggt tgccctgaac tgggggttgg ggggagcgca gcaaaatggc
     4741 ggctgttccc gagtcttgaa tggaagacgc ttgtgaggcg ggctgtgagg tcgttgaaac
     4801 aaggtggggg gcatggtggg cggcaagaac ccaaggtctt gagcccttcg ctaatgcggg
     4861 aaagctctta ttcgggtgag atgggctggg caccatctgg ggaccctgac gtgaagtttg
     4921 tcactgactg gagaactcgg tttgtcgtct gttgcggggg cggcagttat ggcggtgccg
     4981 ttgggcagtg cacccgtacc tttgggagcg cgcgccctcg tcgtgtcgtg acgtcacccg
     5041 ttctgttggc ttataatgca gggtggggcc acctgccggt aggtgtgcgg taggcttttc
     5101 tccgtcgcag gacgcagggt tcgggcctag ggtaggctct cctgaatcga caggcgccgg
     5161 acctctggtg aggggaggga taagtgaggc gtcagtttct ttggtcggtt ttatgtacct
     5221 atcttcttaa gtagctgaag ctccggtttt gaactatgcg ctcggggttg gcgagtgtgt
     5281 tttgtgaagt tttttaggca ccttttgaaa tgtaatcatt tgggtcaata tgtaattttc
     5341 agtgttagac ttgtaaattg tccgctaaat tctggccgtt tttggctttt ttgttagaca
     5401 acatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg
     5461 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg
     5521 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg
     5581 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg
     5641 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg
     5701 gttcccggct ggccgcgcag caacagatgg agggcctcct ggcgccgcac cggcccaagg
     5761 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg
     5821 gcagcgccgt cgtgctcccc ggagtggagg ccgccgagcg cgccggggtg cccgccttcc
     5881 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg
     5941 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct
     6001 aggctagtat gtaagcctag tcttagataa taaaatcgct atccatcgaa gatggatgtg
     6061 tgttggtttt ttgtgtgtgt aacgctaggc gcgcctggtg taccgagaac gatcctctca
     6121 gtgcgagtct cgacgatcca tatcgttgct tggcagtcag ccagtcggaa tccagcttgg
     6181 gacccaggaa gtccaatcgt cagatattgt actcaagcct ggtcacggca gcgtaccgat
     6241 ctgtttaaac ctagatattg atagtctgat cggtcaacgt ataatcgagt cctagctttt
     6301 gcaaacatct atcaagagac aggatcagca ggaggctttc gcatgagtat tcaacatttc
     6361 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     6421 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cgcgagtggg ttacatcgaa
     6481 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ctttccaatg
     6541 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     6601 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ttcaccagtc
     6661 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     6721 atgagtgata acactgcggc caacttactt ctgacaacga ttggaggacc gaaggagcta
     6781 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     6841 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     6901 accttgcgta aactattaac tggcgaacta cttactctag cttcccggca acagttgata
     6961 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     7021 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     7081 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     7141 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     7201 taaccgattc taggtgcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta
     7261 tactcccaca tatgccagat tcagcaacgg atacggcttc cccaacttgc ccacttccat
     7321 acgtgtcctc cttaccagaa atttatcctt aagatcccga atcgtttaaa ctcgactctg
     7381 gctctatcga atctccgtcg tttcgagctt acgcgaac
//
