LOCUS       sequence-306185-        8373 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 8373)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8373)
  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-m4134562
FEATURES             Location/Qualifiers
     source          1..8373
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    94..165
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(170..216)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    234..282
                     /note="synthetic polyadenylation signal"
     enhancer        363..666
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        668..870
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     821..841
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2202..2222
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2256..2333
                     /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             2361..2381
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    2846..2967
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2857..2876)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2911..2930
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      3082..3128
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      3158..3294
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3339..3410
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3636..4224)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3716..3735)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4724..5933
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5770..5789
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5937..6965
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    6992..7040
                     /note="synthetic polyadenylation signal"
     CDS             join(7306..7374,7375..8166)
                     /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(7524..7543)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      8179..8315
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 acgcgaacag ccgtggcgct catttgctcg tcgggcatcg aatctcgtca gctatcgtca
       61 gcttaccttt ttggcagcga tcgcggctcc cgacatcttg gaccattagc tccacaggta
      121 tcttcttccc tctagtggtc ataacagcag cttcagctac ctctcaattc aaaaaacccc
      181 tcaagacccg tttagaggcc ccaaggggtt atgctatcaa tcgttgcgtt acacacacaa
      241 aaaaccaaca cacatccatc ttcgatggat agcgatttta ttatctaact gctgatcgag
      301 tgtagccaga tctagtaatc aattacgggg tcattagttc atagcccata tatggagttc
      361 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      421 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      481 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      541 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      601 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      661 accatgctga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
      721 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
      781 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
      841 gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat cagatctttg
      901 tcgatcctac catccactcg acacacccgc cagcggccgc caccatgtct gataatggac
      961 cacagaacca acgcaatgca ccccgaatca cttttggcgg tccgtccgac agtaccggga
     1021 gcaaccagaa cggcgaaaga tcgggagctc gttcaaaaca gcggaggcct caagggttac
     1081 ccaataacac ggccagctgg ttcacagcgt tgacccagca tggcaaggag gacctcaagt
     1141 tccctcgggg tcagggggtc ccaataaata caaacagttc ccctgatgac caaattggat
     1201 actatagacg cgctactagg aggatccgtg gcggggatgg aaaaatgaag gacctttctc
     1261 ccagatggta cttttattac ctgggtactg gcccagaggc cggtctaccg tatggagcaa
     1321 acaaagatgg cattatttgg gtggccacgg aaggagcact gaatacccca aaggatcaca
     1381 tcggcacccg gaatccagcc aacaatgctg ctatcgttct gcaactcccc caggggacaa
     1441 cattgcctaa aggcttctat gccgagggca gtagaggtgg cagccaggcg tcctcacgca
     1501 gctcttctcg atctcggaac agctcaagaa attccactcc cgggagttcc cgaggaacca
     1561 gcccagcaag gatggctggc aatggtggag atgccgctct ggccctactt ctgctggacc
     1621 gcctcaatca gttagaatca aagatgtcgg ggaaggggca gcaacagcag ggacagaccg
     1681 taactaaaaa gagtgcagcg gaggcctcta aaaagcctcg gcagaaacgt acggcaacaa
     1741 aggcttacaa cgtgacccag gcctttggcc gtagaggacc cgaacagaca caagggaact
     1801 ttggtgacca agagttgata aggcagggaa ccgactacaa acactggcct cagatcgccc
     1861 aattcgcacc gagcgcgagc gcatttttcg gaatgtcacg aattggcatg gaggtgacac
     1921 cttcgggaac ctggctgaca tacactggcg ccataaagct cgatgacaaa gatcccaact
     1981 tcaaggatca agtcatcttg ctgaataagc atattgacgc gtataaaact tttccaccca
     2041 cggaaccgaa gaaggacaag aaaaagaaag ctgatgagac acaggcgctt ccacaaaggc
     2101 agaaaaagca gcagacggtt actctactgc ccgccgcaga cctagatgac ttctccaaac
     2161 agcttcagca gtctatgagt agcgccgact caacccaagc tgaaaacctt tactttcagt
     2221 cgggacggcg aatcccgggg ctgataaatc cctggaaaag gagatggaag aagaacttta
     2281 ttgcagtgtc tgcggctaac cggttcaaaa agatcagttc aagcggcgcc ctagattacg
     2341 acattcctac gaccgcctcc gagaatctct atttccaggg tgaatttggc ttggcacaac
     2401 atgacgaggc cgtcgataac aagtttaata aagaacagca gaacgctttc tatgagatcc
     2461 ttcacttacc aaatctgaac gaagagcaac gcaacgcttt cattcagagc ctgaaagacg
     2521 atccctcgca gtctgcaaat ctcctggccg aagccaagaa gctgaatgac gcacaggctc
     2581 caaaagttga taacaagttt aacaaggagc agcagaatgc tttctacgag atattgcacc
     2641 ttcctaatct gaatgaggaa caaagaaatg catttatcca gtccctaaaa gacgatccat
     2701 cacagagtgc caacctgctt gcggaagcca aaaagctcaa tgatgcccag gccccgaaag
     2761 tagacgcgaa ccatcaataa ggatccgcag gcctctgcta gcttgactga ctgagataca
     2821 gcgtaccttc agctcacaga catgataaga tacattgatg agtttggaca aaccacaact
     2881 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     2941 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     3001 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatt
     3061 ggcccatctc tatcggtatc gtagcataac cccttggggc ctctaaacgg gtcttgaggg
     3121 gttttttgtg cccctcgggc cggattgcta tctaccggca ttggcgcaga aaaaaatgcc
     3181 tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac ggatacggct
     3241 tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc ttaaggtcgt
     3301 cagctatcct gcaggcgatc tctcgatttc gatcaagaca ttcctttaat ggtcttttct
     3361 ggacaccact aggggtcaga agtagttcat caaactttct tccctcccta atctcattgg
     3421 ttaccttggg ctatcgaaac ttaattaacc agtcaagtca gctacttggc gagatcgact
     3481 tgtctgggtt tcgactacgc tcagaattgc gtcagtcaag ttcgatctgg tccttgctat
     3541 tgcacccgtt ctccgattac gagtttcatt taaatcatgt gagcaaaagg ccagcaaaag
     3601 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3661 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3721 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3781 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3841 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3901 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3961 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4021 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     4081 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4141 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4201 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     4261 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     4321 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     4381 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     4441 tttcgttcat ccatagttgc atttaaattt ccgaactctc caaggccctc gtcggaaaat
     4501 cttcaaacct ttcgtccgat ccatcttgca ggctacctct cgaacgaact atcgcaagtc
     4561 tcttggccgg ccttgcgcct tggctattgc ttggcagcgc ctatcgccag gtattactcc
     4621 aatcccgaat atccgagatc gggatcaccc gagagaagtt caacctacat cctcaatccc
     4681 gatctatccg agatccgagg aatatcgaaa tcggggcgcg cctggcctcc gcgccgggtt
     4741 ttggcgcctc ccgcgggcgc ccccctcgtc acggcgagcg ctgccacgtc agacgaaggg
     4801 cgcaggagcg tcctgatcct tccgcccgga cgctcaggac agcggcccgc tgctcataag
     4861 actcggcctt agaaccccag tatcagcaga aggacatttt aggacgggac ttgggtgact
     4921 ctagggcact ggttttcttt ccagagagcg gaacaggcga ggaaaagtag tcccttctcg
     4981 gcgattctgc ggagggatct ccgtggggcg gtgaacgccg atgattatat aaggacgcgc
     5041 cgggtgtggc acagctagtt ccgtcgcagc cgggatttgg gtcgcggttc ttgtttgtgg
     5101 atcgctgtga tcgtcacttg gtgagtagcg ggctgctggg ctggccgggg ctttcgtggc
     5161 cgccgggccg ctcggtggga cggaagcgtg tggagagacc gccaagggct gtagtctggg
     5221 tccgcgagca aggttgccct gaactggggg ttggggggag cgcagcaaaa tggcggctgt
     5281 tcccgagtct tgaatggaag acgcttgtga ggcgggctgt gaggtcgttg aaacaaggtg
     5341 gggggcatgg tgggcggcaa gaacccaagg tcttgagccc ttcgctaatg cgggaaagct
     5401 cttattcggg tgagatgggc tgggcaccat ctggggaccc tgacgtgaag tttgtcactg
     5461 actggagaac tcggtttgtc gtctgttgcg ggggcggcag ttatggcggt gccgttgggc
     5521 agtgcacccg tacctttggg agcgcgcgcc ctcgtcgtgt cgtgacgtca cccgttctgt
     5581 tggcttataa tgcagggtgg ggccacctgc cggtaggtgt gcggtaggct tttctccgtc
     5641 gcaggacgca gggttcgggc ctagggtagg ctctcctgaa tcgacaggcg ccggacctct
     5701 ggtgagggga gggataagtg aggcgtcagt ttctttggtc ggttttatgt acctatcttc
     5761 ttaagtagct gaagctccgg ttttgaacta tgcgctcggg gttggcgagt gtgttttgtg
     5821 aagtttttta ggcacctttt gaaatgtaat catttgggtc aatatgtaat tttcagtgtt
     5881 agacttgtaa attgtccgct aaattctggc cgtttttggc ttttttgtta gacaacatgg
     5941 gtaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca
     6001 gcgtctccga cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg
     6061 taggagggcg tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc
     6121 gttatgttta tcggcacttt gcatccgccg cgctcccgat tccggaagtg cttgacattg
     6181 gggagttcag cgagagcctg acctattgca tctcccgccg tgcacagggt gtcacgttgc
     6241 aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag gcaatggatg
     6301 ccatcgctgc cgccgatctt agccagacga gcgggttcgg cccattcgga ccgcaaggaa
     6361 tcggtcaata cactacatgg cgtgatttca tatgcgcgat tgctgatccc catgtgtatc
     6421 actggcaaac tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc
     6481 tgatgctttg ggccgaggac tgccccgaag tccggcacct cgtgcacgcg gatttcggct
     6541 ccaacaatgt cctgacggac aatggccgca taacagcggt cattgactgg agcgaggcga
     6601 tgttcgggga ttcccaatac gaggtcgcca acatcttctt ctggaggccg tggttggctt
     6661 gtatggagca gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgcccc
     6721 ggctccgggc gtatatgctc cgcattggtc ttgaccaact ctatcagagc ttggttgacg
     6781 gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag
     6841 ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc cgccgtctgg accgatggct
     6901 gtgtagaagt actcgccgat agtggaaacc gacgccccag cactcgtccg agggcaaagg
     6961 aataagctag tatgtaagcc tagtcttaga taataaaatc gctatccatc gaagatggat
     7021 gtgtgttggt tttttgtgtg tgtaacgcta ggcgcgcctg gtgtaccgag aacgatcctc
     7081 tcagtgcgag tctcgacgat ccatatcgtt gcttggcagt cagccagtcg gaatccagct
     7141 tgggacccag gaagtccaat cgtcagatat tgtactcaag cctggtcacg gcagcgtacc
     7201 gatctgttta aacctagata ttgatagtct gatcggtcaa cgtataatcg agtcctagct
     7261 tttgcaaaca tctatcaaga gacaggatca gcaggaggct ttcgcatgag tattcaacat
     7321 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     7381 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcgcgagt gggttacatc
     7441 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgctttcca
     7501 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     7561 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtattcacca
     7621 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     7681 accatgagtg ataacactgc ggccaactta cttctgacaa cgattggagg accgaaggag
     7741 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     7801 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     7861 acaaccttgc gtaaactatt aactggcgaa ctacttactc tagcttcccg gcaacagttg
     7921 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     7981 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     8041 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     8101 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     8161 tggtaaccga ttctaggtgc attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc
     8221 ttatactccc acatatgcca gattcagcaa cggatacggc ttccccaact tgcccacttc
     8281 catacgtgtc ctccttacca gaaatttatc cttaagatcc cgaatcgttt aaactcgact
     8341 ctggctctat cgaatctccg tcgtttcgag ctt
//
