LOCUS       sequence-306085-        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-m4134501
FEATURES             Location/Qualifiers
     source          1..7928
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    134..205
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(210..256)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    274..322
                     /note="synthetic polyadenylation signal"
     enhancer        403..706
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        708..910
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     861..881
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2242..2262
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2296..2373
                     /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             2401..2421
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    2870..2991
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2881..2900)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2935..2954
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      3106..3152
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      3182..3318
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3363..3434
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3660..4248)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3740..3759)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4748..5957
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5794..5813
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5961..6560
                     /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(5961..5980)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6457..6477
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    6587..6635
                     /note="synthetic polyadenylation signal"
     CDS             join(6901..6969,6970..7761)
                     /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(7119..7138)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7774..7910
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 ctcgactctg gctctatcga atctccgtcg tttcgagctt acgcgaacag ccgtggcgct
       61 catttgctcg tcgggcatcg aatctcgtca gctatcgtca gcttaccttt ttggcagcga
      121 tcgcggctcc cgacatcttg gaccattagc tccacaggta tcttcttccc tctagtggtc
      181 ataacagcag cttcagctac ctctcaattc aaaaaacccc tcaagacccg tttagaggcc
      241 ccaaggggtt atgctatcaa tcgttgcgtt acacacacaa aaaaccaaca cacatccatc
      301 ttcgatggat agcgatttta ttatctaact gctgatcgag tgtagccaga tctagtaatc
      361 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
      421 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      481 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      541 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      601 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      661 tcctacttgg cagtacatct acgtattagt catcgctatt accatgctga tgcggttttg
      721 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
      781 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
      841 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
      901 aagcagagct ggtttagtga accgtcagat cagatctttg tcgatcctac catccactcg
      961 acacacccgc cagcggccgc caccatgtct gataatggac cacagaacca acgcaatgca
     1021 ccccgaatca cttttggcgg tccgtccgac agtaccggga gcaaccagaa cggcgaaaga
     1081 tcgggagctc gttcaaaaca gcggaggcct caagggttac ccaataacac ggccagctgg
     1141 ttcacagcgt tgacccagca tggcaaggag gacctcaagt tccctcgggg tcagggggtc
     1201 ccaataaata caaacagttc ccctgatgac caaattggat actatagacg cgctactagg
     1261 aggatccgtg gcggggatgg aaaaatgaag gacctttctc ccagatggta cttttattac
     1321 ctgggtactg gcccagaggc cggtctaccg tatggagcaa acaaagatgg cattatttgg
     1381 gtggccacgg aaggagcact gaatacccca aaggatcaca tcggcacccg gaatccagcc
     1441 aacaatgctg ctatcgttct gcaactcccc caggggacaa cattgcctaa aggcttctat
     1501 gccgagggca gtagaggtgg cagccaggcg tcctcacgca gctcttctcg atctcggaac
     1561 agctcaagaa attccactcc cgggagttcc cgaggaacca gcccagcaag gatggctggc
     1621 aatggtggag atgccgctct ggccctactt ctgctggacc gcctcaatca gttagaatca
     1681 aagatgtcgg ggaaggggca gcaacagcag ggacagaccg taactaaaaa gagtgcagcg
     1741 gaggcctcta aaaagcctcg gcagaaacgt acggcaacaa aggcttacaa cgtgacccag
     1801 gcctttggcc gtagaggacc cgaacagaca caagggaact ttggtgacca agagttgata
     1861 aggcagggaa ccgactacaa acactggcct cagatcgccc aattcgcacc gagcgcgagc
     1921 gcatttttcg gaatgtcacg aattggcatg gaggtgacac cttcgggaac ctggctgaca
     1981 tacactggcg ccataaagct cgatgacaaa gatcccaact tcaaggatca agtcatcttg
     2041 ctgaataagc atattgacgc gtataaaact tttccaccca cggaaccgaa gaaggacaag
     2101 aaaaagaaag ctgatgagac acaggcgctt ccacaaaggc agaaaaagca gcagacggtt
     2161 actctactgc ccgccgcaga cctagatgac ttctccaaac agcttcagca gtctatgagt
     2221 agcgccgact caacccaagc tgaaaacctt tactttcagt cgggacggcg aatcccgggg
     2281 ctgataaatc cctggaaaag gagatggaag aagaacttta ttgcagtgtc tgcggctaac
     2341 cggttcaaaa agatcagttc aagcggcgcc ctagattacg acattcctac gaccgcctcc
     2401 gagaatctct atttccaggg tgaatttggc ttggcacaac atgacgaggc cgtcgataac
     2461 aagtttaata aagaacagca gaacgctttc tatgagatcc ttcacttacc aaatctgaac
     2521 gaagagcaac gcaacgcttt cattcagagc ctgaaagacg atccctcgca gtctgcaaat
     2581 ctcctggccg aagccaagaa gctgaatgac gcacaggctc caaaagttga taacaagttt
     2641 aacaaggagc agcagaatgc tttctacgag atattgcacc ttcctaatct gaatgaggaa
     2701 caaagaaatg catttatcca gtccctaaaa gacgatccat cacagagtgc caacctgctt
     2761 gcggaagcca aaaagctcaa tgatgcccag gccccgaaag tagacgcgaa ccatcaataa
     2821 gctagcttga ctgactgaga tacagcgtac cttcagctca cagacatgat aagatacatt
     2881 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     2941 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     3001 aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag
     3061 taaaacctct acaaatgtgg tattggccca tctctatcgg tatcgtagca taaccccttg
     3121 gggcctctaa acgggtcttg aggggttttt tgtgcccctc gggccggatt gctatctacc
     3181 ggcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact cccacatatg
     3241 ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt gtcctcctta
     3301 ccagaaattt atccttaagg tcgtcagcta tcctgcaggc gatctctcga tttcgatcaa
     3361 gacattcctt taatggtctt ttctggacac cactaggggt cagaagtagt tcatcaaact
     3421 ttcttccctc cctaatctca ttggttacct tgggctatcg aaacttaatt aaccagtcaa
     3481 gtcagctact tggcgagatc gacttgtctg ggtttcgact acgctcagaa ttgcgtcagt
     3541 caagttcgat ctggtccttg ctattgcacc cgttctccga ttacgagttt catttaaatc
     3601 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     3661 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     3721 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     3781 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     3841 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     3901 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     3961 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     4021 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     4081 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     4141 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     4201 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     4261 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     4321 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     4381 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     4441 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcatttaa atttccgaac
     4501 tctccaaggc cctcgtcgga aaatcttcaa acctttcgtc cgatccatct tgcaggctac
     4561 ctctcgaacg aactatcgca agtctcttgg ccggccttgc gccttggcta ttgcttggca
     4621 gcgcctatcg ccaggtatta ctccaatccc gaatatccga gatcgggatc acccgagaga
     4681 agttcaacct acatcctcaa tcccgatcta tccgagatcc gaggaatatc gaaatcgggg
     4741 cgcgcctggc ctccgcgccg ggttttggcg cctcccgcgg gcgcccccct cgtcacggcg
     4801 agcgctgcca cgtcagacga agggcgcagg agcgtcctga tccttccgcc cggacgctca
     4861 ggacagcggc ccgctgctca taagactcgg ccttagaacc ccagtatcag cagaaggaca
     4921 ttttaggacg ggacttgggt gactctaggg cactggtttt ctttccagag agcggaacag
     4981 gcgaggaaaa gtagtccctt ctcggcgatt ctgcggaggg atctccgtgg ggcggtgaac
     5041 gccgatgatt atataaggac gcgccgggtg tggcacagct agttccgtcg cagccgggat
     5101 ttgggtcgcg gttcttgttt gtggatcgct gtgatcgtca cttggtgagt agcgggctgc
     5161 tgggctggcc ggggctttcg tggccgccgg gccgctcggt gggacggaag cgtgtggaga
     5221 gaccgccaag ggctgtagtc tgggtccgcg agcaaggttg ccctgaactg ggggttgggg
     5281 ggagcgcagc aaaatggcgg ctgttcccga gtcttgaatg gaagacgctt gtgaggcggg
     5341 ctgtgaggtc gttgaaacaa ggtggggggc atggtgggcg gcaagaaccc aaggtcttga
     5401 gcccttcgct aatgcgggaa agctcttatt cgggtgagat gggctgggca ccatctgggg
     5461 accctgacgt gaagtttgtc actgactgga gaactcggtt tgtcgtctgt tgcgggggcg
     5521 gcagttatgg cggtgccgtt gggcagtgca cccgtacctt tgggagcgcg cgccctcgtc
     5581 gtgtcgtgac gtcacccgtt ctgttggctt ataatgcagg gtggggccac ctgccggtag
     5641 gtgtgcggta ggcttttctc cgtcgcagga cgcagggttc gggcctaggg taggctctcc
     5701 tgaatcgaca ggcgccggac ctctggtgag gggagggata agtgaggcgt cagtttcttt
     5761 ggtcggtttt atgtacctat cttcttaagt agctgaagct ccggttttga actatgcgct
     5821 cggggttggc gagtgtgttt tgtgaagttt tttaggcacc ttttgaaatg taatcatttg
     5881 ggtcaatatg taattttcag tgttagactt gtaaattgtc cgctaaattc tggccgtttt
     5941 tggctttttt gttagacaac atgaccgagt acaagcccac ggtgcgcctc gccacccgcg
     6001 acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc
     6061 gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc
     6121 tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg
     6181 cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc
     6241 gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggag ggcctcctgg
     6301 cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc
     6361 accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcc gccgagcgcg
     6421 ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc
     6481 tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga
     6541 cccgcaagcc cggtgcctag gctagtatgt aagcctagtc ttagataata aaatcgctat
     6601 ccatcgaaga tggatgtgtg ttggtttttt gtgtgtgtaa cgctaggcgc gcctggtgta
     6661 ccgagaacga tcctctcagt gcgagtctcg acgatccata tcgttgcttg gcagtcagcc
     6721 agtcggaatc cagcttggga cccaggaagt ccaatcgtca gatattgtac tcaagcctgg
     6781 tcacggcagc gtaccgatct gtttaaacct agatattgat agtctgatcg gtcaacgtat
     6841 aatcgagtcc tagcttttgc aaacatctat caagagacag gatcagcagg aggctttcgc
     6901 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     6961 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgcg
     7021 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     7081 gaagaacgct ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     7141 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     7201 gttgagtatt caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     7261 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatt
     7321 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     7381 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     7441 cctgtagcaa tggcaacaac cttgcgtaaa ctattaactg gcgaactact tactctagct
     7501 tcccggcaac agttgataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     7561 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     7621 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     7681 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     7741 tcactgatta agcattggta accgattcta ggtgcattgg cgcagaaaaa aatgcctgat
     7801 gcgacgctgc gcgtcttata ctcccacata tgccagattc agcaacggat acggcttccc
     7861 caacttgccc acttccatac gtgtcctcct taccagaaat ttatccttaa gatcccgaat
     7921 cgtttaaa
//
