LOCUS       sequence-338674-        7896 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 7896)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7896)
  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-m4134312
FEATURES             Location/Qualifiers
     source          1..7896
                     /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             1861..1938
                     /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"
     polyA_signal    2007..2128
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2018..2037)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2072..2091
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2243..2289
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2319..2455
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2500..2571
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2797..3385)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2877..2896)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3885..5094
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4931..4950
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5098..6126
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    6153..6201
                     /note="synthetic polyadenylation signal"
     CDS             join(6467..6535,6536..7327)
                     /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(6685..6704)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7340..7476
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7628..7699
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7704..7750)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7768..7816
                     /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 aagaggcgat ggaaaaagaa ttttatcgcg gtgagtgcag ccaacagatt caaaaagata
     1921 tcctcgtctg gtgccctcta aggatccgca ggcctctgct agcttgactg actgagatac
     1981 agcgtacctt cagctcacag acatgataag atacattgat gagtttggac aaaccacaac
     2041 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     2101 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     2161 ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat
     2221 tggcccatct ctatcggtat cgtagcataa ccccttgggg cctctaaacg ggtcttgagg
     2281 ggttttttgt gcccctcggg ccggattgct atctaccggc attggcgcag aaaaaaatgc
     2341 ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc
     2401 ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaaggtcg
     2461 tcagctatcc tgcaggcgat ctctcgattt cgatcaagac attcctttaa tggtcttttc
     2521 tggacaccac taggggtcag aagtagttca tcaaactttc ttccctccct aatctcattg
     2581 gttaccttgg gctatcgaaa cttaattaac cagtcaagtc agctacttgg cgagatcgac
     2641 ttgtctgggt ttcgactacg ctcagaattg cgtcagtcaa gttcgatctg gtccttgcta
     2701 ttgcacccgt tctccgatta cgagtttcat ttaaatcatg tgagcaaaag gccagcaaaa
     2761 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2821 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2881 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2941 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3001 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3061 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3121 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3181 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     3241 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3301 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3361 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3421 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3481 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3541 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     3601 atttcgttca tccatagttg catttaaatt tccgaactct ccaaggccct cgtcggaaaa
     3661 tcttcaaacc tttcgtccga tccatcttgc aggctacctc tcgaacgaac tatcgcaagt
     3721 ctcttggccg gccttgcgcc ttggctattg cttggcagcg cctatcgcca ggtattactc
     3781 caatcccgaa tatccgagat cgggatcacc cgagagaagt tcaacctaca tcctcaatcc
     3841 cgatctatcc gagatccgag gaatatcgaa atcggggcgc gcctggcctc cgcgccgggt
     3901 tttggcgcct cccgcgggcg cccccctcgt cacggcgagc gctgccacgt cagacgaagg
     3961 gcgcaggagc gtcctgatcc ttccgcccgg acgctcagga cagcggcccg ctgctcataa
     4021 gactcggcct tagaacccca gtatcagcag aaggacattt taggacggga cttgggtgac
     4081 tctagggcac tggttttctt tccagagagc ggaacaggcg aggaaaagta gtcccttctc
     4141 ggcgattctg cggagggatc tccgtggggc ggtgaacgcc gatgattata taaggacgcg
     4201 ccgggtgtgg cacagctagt tccgtcgcag ccgggatttg ggtcgcggtt cttgtttgtg
     4261 gatcgctgtg atcgtcactt ggtgagtagc gggctgctgg gctggccggg gctttcgtgg
     4321 ccgccgggcc gctcggtggg acggaagcgt gtggagagac cgccaagggc tgtagtctgg
     4381 gtccgcgagc aaggttgccc tgaactgggg gttgggggga gcgcagcaaa atggcggctg
     4441 ttcccgagtc ttgaatggaa gacgcttgtg aggcgggctg tgaggtcgtt gaaacaaggt
     4501 ggggggcatg gtgggcggca agaacccaag gtcttgagcc cttcgctaat gcgggaaagc
     4561 tcttattcgg gtgagatggg ctgggcacca tctggggacc ctgacgtgaa gtttgtcact
     4621 gactggagaa ctcggtttgt cgtctgttgc gggggcggca gttatggcgg tgccgttggg
     4681 cagtgcaccc gtacctttgg gagcgcgcgc cctcgtcgtg tcgtgacgtc acccgttctg
     4741 ttggcttata atgcagggtg gggccacctg ccggtaggtg tgcggtaggc ttttctccgt
     4801 cgcaggacgc agggttcggg cctagggtag gctctcctga atcgacaggc gccggacctc
     4861 tggtgagggg agggataagt gaggcgtcag tttctttggt cggttttatg tacctatctt
     4921 cttaagtagc tgaagctccg gttttgaact atgcgctcgg ggttggcgag tgtgttttgt
     4981 gaagtttttt aggcaccttt tgaaatgtaa tcatttgggt caatatgtaa ttttcagtgt
     5041 tagacttgta aattgtccgc taaattctgg ccgtttttgg cttttttgtt agacaacatg
     5101 ggtaaaaagc ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac
     5161 agcgtctccg acctgatgca gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat
     5221 gtaggagggc gtggatatgt cctgcgggta aatagctgcg ccgatggttt ctacaaagat
     5281 cgttatgttt atcggcactt tgcatccgcc gcgctcccga ttccggaagt gcttgacatt
     5341 ggggagttca gcgagagcct gacctattgc atctcccgcc gtgcacaggg tgtcacgttg
     5401 caagacctgc ctgaaaccga actgcccgct gttctgcagc cggtcgcgga ggcaatggat
     5461 gccatcgctg ccgccgatct tagccagacg agcgggttcg gcccattcgg accgcaagga
     5521 atcggtcaat acactacatg gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat
     5581 cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag
     5641 ctgatgcttt gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc
     5701 tccaacaatg tcctgacgga caatggccgc ataacagcgg tcattgactg gagcgaggcg
     5761 atgttcgggg attcccaata cgaggtcgcc aacatcttct tctggaggcc gtggttggct
     5821 tgtatggagc agcagacgcg ctacttcgag cggaggcatc cggagcttgc aggatcgccc
     5881 cggctccggg cgtatatgct ccgcattggt cttgaccaac tctatcagag cttggttgac
     5941 ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga
     6001 gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg ccgccgtctg gaccgatggc
     6061 tgtgtagaag tactcgccga tagtggaaac cgacgcccca gcactcgtcc gagggcaaag
     6121 gaataagcta gtatgtaagc ctagtcttag ataataaaat cgctatccat cgaagatgga
     6181 tgtgtgttgg ttttttgtgt gtgtaacgct aggcgcgcct ggtgtaccga gaacgatcct
     6241 ctcagtgcga gtctcgacga tccatatcgt tgcttggcag tcagccagtc ggaatccagc
     6301 ttgggaccca ggaagtccaa tcgtcagata ttgtactcaa gcctggtcac ggcagcgtac
     6361 cgatctgttt aaacctagat attgatagtc tgatcggtca acgtataatc gagtcctagc
     6421 ttttgcaaac atctatcaag agacaggatc agcaggaggc tttcgcatga gtattcaaca
     6481 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     6541 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcgcgag tgggttacat
     6601 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgctttcc
     6661 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     6721 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtattcacc
     6781 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     6841 aaccatgagt gataacactg cggccaactt acttctgaca acgattggag gaccgaagga
     6901 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     6961 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     7021 aacaaccttg cgtaaactat taactggcga actacttact ctagcttccc ggcaacagtt
     7081 gatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     7141 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     7201 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     7261 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     7321 ttggtaaccg attctaggtg cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt
     7381 cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac ttgcccactt
     7441 ccatacgtgt cctccttacc agaaatttat ccttaagatc ccgaatcgtt taaactcgac
     7501 tctggctcta tcgaatctcc gtcgtttcga gcttacgcga acagccgtgg cgctcatttg
     7561 ctcgtcgggc atcgaatctc gtcagctatc gtcagcttac ctttttggca gcgatcgcgg
     7621 ctcccgacat cttggaccat tagctccaca ggtatcttct tccctctagt ggtcataaca
     7681 gcagcttcag ctacctctca attcaaaaaa cccctcaaga cccgtttaga ggccccaagg
     7741 ggttatgcta tcaatcgttg cgttacacac acaaaaaacc aacacacatc catcttcgat
     7801 ggatagcgat tttattatct aactgctgat cgagtgtagc cagatctagt aatcaattac
     7861 ggggtcatta gttcatagcc catatatgga gttccg
//
