LOCUS       sequence-300241-        7928 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 7928)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7928)
  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-m4133833
FEATURES             Location/Qualifiers
     source          1..7928
                     /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             1894..1971
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             1999..2019
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    2468..2589
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2479..2498)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2533..2552
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2704..2750
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2780..2916
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2961..3032
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3258..3846)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3338..3357)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4346..5555
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5392..5411
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5559..6158
                     /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(5559..5578)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6055..6075
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    6185..6233
                     /note="synthetic polyadenylation signal"
     CDS             join(6499..6567,6568..7359)
                     /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(6717..6736)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7372..7508
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7660..7731
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7736..7782)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7800..7848
                     /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 taatggaccc
      601 caaaatcagc gaaatgcacc ccgcattacg tttggtggac cctcagattc aactggcagt
      661 aaccagaatg gagaacgcag tggggcgcga tcaaaacaac gtcggcccca aggtttaccc
      721 aataatactg cgtcttggtt caccgctctc actcaacatg gcaaggaaga ccttaaattc
      781 cctcgaggac aaggcgttcc aattaacacc aatagcagtc cagatgacca aattggctac
      841 taccgaagag ctaccagacg aattcgtggt ggtgacggta aaatgaaaga tctcagtcca
      901 agatggtatt tctactacct aggaactggg ccagaagctg gacttcccta tggtgctaac
      961 aaagacggca tcatatgggt tgcaactgag ggagccttga atacaccaaa agatcacatt
     1021 ggcacccgca atcctgctaa caatgctgca atcgtgctac aacttcctca aggaacaaca
     1081 ttgccaaaag gcttctacgc agaagggagc agaggcggca gtcaagcctc ttctcgttcc
     1141 tcatcacgta gtcgcaacag ttcaagaaat tcaactccag gcagcagtag gggaacttct
     1201 cctgctagaa tggctggcaa tggcggtgat gctgctcttg ctttgctgct gcttgacaga
     1261 ttgaaccagc ttgagagcaa aatgtctggt aaaggccaac aacaacaagg ccaaactgtc
     1321 actaagaaat ctgctgctga ggcttctaag aagcctcggc aaaaacgtac tgccactaaa
     1381 gcatacaatg taacacaagc tttcggcaga cgtggtccag aacaaaccca aggaaatttt
     1441 ggggaccagg aactaatcag acaaggaact gattacaaac attggccgca aattgcacaa
     1501 tttgccccca gcgcttcagc gttcttcgga atgtcgcgca ttggcatgga agtcacacct
     1561 tcgggaacgt ggttgaccta cacaggtgcc atcaaattgg atgacaaaga tccaaatttc
     1621 aaagatcaag tcattttgct gaataagcat attgacgcat acaaaacatt cccaccaaca
     1681 gagcctaaaa aggacaaaaa gaagaaggct gatgaaactc aagccttacc gcagagacag
     1741 aagaaacagc aaactgtgac tcttcttcct gctgcagatt tggatgattt ctccaaacaa
     1801 ttgcaacaat ccatgagcag tgctgactca actcaggccg aaaaccttta ctttcagtcg
     1861 ggacggcgaa tcccggggct gataaatccc tggaaaagga gatggaagaa gaactttatt
     1921 gcagtgtctg cggctaaccg gttcaaaaag atcagttcaa gcggcgccct agattacgac
     1981 attcctacga ccgcctccga gaatctctat ttccagggtg aatttggctt ggcacaacat
     2041 gacgaggccg tcgataacaa gtttaataaa gaacagcaga acgctttcta tgagatcctt
     2101 cacttaccaa atctgaacga agagcaacgc aacgctttca ttcagagcct gaaagacgat
     2161 ccctcgcagt ctgcaaatct cctggccgaa gccaagaagc tgaatgacgc acaggctcca
     2221 aaagttgata acaagtttaa caaggagcag cagaatgctt tctacgagat attgcacctt
     2281 cctaatctga atgaggaaca aagaaatgca tttatccagt ccctaaaaga cgatccatca
     2341 cagagtgcca acctgcttgc ggaagccaaa aagctcaatg atgcccaggc cccgaaagta
     2401 gacgcgaacc atcaataagc tagcttgact gactgagata cagcgtacct tcagctcaca
     2461 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     2521 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     2581 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg
     2641 gaggtttttt aaagcaagta aaacctctac aaatgtggta ttggcccatc tctatcggta
     2701 tcgtagcata accccttggg gcctctaaac gggtcttgag gggttttttg tgcccctcgg
     2761 gccggattgc tatctaccgg cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt
     2821 cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac ttgcccactt
     2881 ccatacgtgt cctccttacc agaaatttat ccttaaggtc gtcagctatc ctgcaggcga
     2941 tctctcgatt tcgatcaaga cattccttta atggtctttt ctggacacca ctaggggtca
     3001 gaagtagttc atcaaacttt cttccctccc taatctcatt ggttaccttg ggctatcgaa
     3061 acttaattaa ccagtcaagt cagctacttg gcgagatcga cttgtctggg tttcgactac
     3121 gctcagaatt gcgtcagtca agttcgatct ggtccttgct attgcacccg ttctccgatt
     3181 acgagtttca tttaaatcat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     3241 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     3301 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     3361 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     3421 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     3481 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     3541 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     3601 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     3661 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     3721 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     3781 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     3841 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     3901 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     3961 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     4021 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     4081 gcatttaaat ttccgaactc tccaaggccc tcgtcggaaa atcttcaaac ctttcgtccg
     4141 atccatcttg caggctacct ctcgaacgaa ctatcgcaag tctcttggcc ggccttgcgc
     4201 cttggctatt gcttggcagc gcctatcgcc aggtattact ccaatcccga atatccgaga
     4261 tcgggatcac ccgagagaag ttcaacctac atcctcaatc ccgatctatc cgagatccga
     4321 ggaatatcga aatcggggcg cgcctggcct ccgcgccggg ttttggcgcc tcccgcgggc
     4381 gcccccctcg tcacggcgag cgctgccacg tcagacgaag ggcgcaggag cgtcctgatc
     4441 cttccgcccg gacgctcagg acagcggccc gctgctcata agactcggcc ttagaacccc
     4501 agtatcagca gaaggacatt ttaggacggg acttgggtga ctctagggca ctggttttct
     4561 ttccagagag cggaacaggc gaggaaaagt agtcccttct cggcgattct gcggagggat
     4621 ctccgtgggg cggtgaacgc cgatgattat ataaggacgc gccgggtgtg gcacagctag
     4681 ttccgtcgca gccgggattt gggtcgcggt tcttgtttgt ggatcgctgt gatcgtcact
     4741 tggtgagtag cgggctgctg ggctggccgg ggctttcgtg gccgccgggc cgctcggtgg
     4801 gacggaagcg tgtggagaga ccgccaaggg ctgtagtctg ggtccgcgag caaggttgcc
     4861 ctgaactggg ggttgggggg agcgcagcaa aatggcggct gttcccgagt cttgaatgga
     4921 agacgcttgt gaggcgggct gtgaggtcgt tgaaacaagg tggggggcat ggtgggcggc
     4981 aagaacccaa ggtcttgagc ccttcgctaa tgcgggaaag ctcttattcg ggtgagatgg
     5041 gctgggcacc atctggggac cctgacgtga agtttgtcac tgactggaga actcggtttg
     5101 tcgtctgttg cgggggcggc agttatggcg gtgccgttgg gcagtgcacc cgtacctttg
     5161 ggagcgcgcg ccctcgtcgt gtcgtgacgt cacccgttct gttggcttat aatgcagggt
     5221 ggggccacct gccggtaggt gtgcggtagg cttttctccg tcgcaggacg cagggttcgg
     5281 gcctagggta ggctctcctg aatcgacagg cgccggacct ctggtgaggg gagggataag
     5341 tgaggcgtca gtttctttgg tcggttttat gtacctatct tcttaagtag ctgaagctcc
     5401 ggttttgaac tatgcgctcg gggttggcga gtgtgttttg tgaagttttt taggcacctt
     5461 ttgaaatgta atcatttggg tcaatatgta attttcagtg ttagacttgt aaattgtccg
     5521 ctaaattctg gccgtttttg gcttttttgt tagacaacat gaccgagtac aagcccacgg
     5581 tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg
     5641 ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg
     5701 agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg
     5761 acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt
     5821 tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac
     5881 agatggaggg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg
     5941 tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag
     6001 tggaggccgc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc
     6061 tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac
     6121 cgcgcacctg gtgcatgacc cgcaagcccg gtgcctaggc tagtatgtaa gcctagtctt
     6181 agataataaa atcgctatcc atcgaagatg gatgtgtgtt ggttttttgt gtgtgtaacg
     6241 ctaggcgcgc ctggtgtacc gagaacgatc ctctcagtgc gagtctcgac gatccatatc
     6301 gttgcttggc agtcagccag tcggaatcca gcttgggacc caggaagtcc aatcgtcaga
     6361 tattgtactc aagcctggtc acggcagcgt accgatctgt ttaaacctag atattgatag
     6421 tctgatcggt caacgtataa tcgagtccta gcttttgcaa acatctatca agagacagga
     6481 tcagcaggag gctttcgcat gagtattcaa catttccgtg tcgcccttat tccctttttt
     6541 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     6601 gaagatcagt tgggtgcgcg agtgggttac atcgaactgg atctcaacag cggtaagatc
     6661 cttgagagtt ttcgccccga agaacgcttt ccaatgatga gcacttttaa agttctgcta
     6721 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     6781 tattctcaga atgacttggt tgagtattca ccagtcacag aaaagcatct tacggatggc
     6841 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     6901 ttacttctga caacgattgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     6961 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     7021 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacct tgcgtaaact attaactggc
     7081 gaactactta ctctagcttc ccggcaacag ttgatagact ggatggaggc ggataaagtt
     7141 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     7201 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     7261 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     7321 atcgctgaga taggtgcctc actgattaag cattggtaac cgattctagg tgcattggcg
     7381 cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag
     7441 caacggatac ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt
     7501 atccttaaga tcccgaatcg tttaaactcg actctggctc tatcgaatct ccgtcgtttc
     7561 gagcttacgc gaacagccgt ggcgctcatt tgctcgtcgg gcatcgaatc tcgtcagcta
     7621 tcgtcagctt acctttttgg cagcgatcgc ggctcccgac atcttggacc attagctcca
     7681 caggtatctt cttccctcta gtggtcataa cagcagcttc agctacctct caattcaaaa
     7741 aacccctcaa gacccgttta gaggccccaa ggggttatgc tatcaatcgt tgcgttacac
     7801 acacaaaaaa ccaacacaca tccatcttcg atggatagcg attttattat ctaactgctg
     7861 atcgagtgta gccagatcta gtaatcaatt acggggtcat tagttcatag cccatatatg
     7921 gagttccg
//
