LOCUS       sequence-300271-        6021 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 6021)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6021)
  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-m4133993
FEATURES             Location/Qualifiers
     source          1..6021
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    62..133
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(138..184)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    202..250
                     /note="synthetic polyadenylation signal"
     enhancer        331..634
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        636..838
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     789..809
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2170..2190
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2224..2301
                     /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             2329..2349
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    2798..2919
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2809..2828)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2863..2882
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      3034..3080
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      3110..3246
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3291..3362
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3588..4176)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3668..3687)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(4922..4990,4991..5782)
                     /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(5140..5159)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5795..5931
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 gggcatcgaa tctcgtcagc tatcgtcagc ttaccttttt ggcagcgatc gcggctcccg
       61 acatcttgga ccattagctc cacaggtatc ttcttccctc tagtggtcat aacagcagct
      121 tcagctacct ctcaattcaa aaaacccctc aagacccgtt tagaggcccc aaggggttat
      181 gctatcaatc gttgcgttac acacacaaaa aaccaacaca catccatctt cgatggatag
      241 cgattttatt atctaactgc tgatcgagtg tagccagatc tagtaatcaa ttacggggtc
      301 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
      361 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
      421 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
      481 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
      541 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
      601 gtacatctac gtattagtca tcgctattac catgctgatg cggttttggc agtacatcaa
      661 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
      721 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
      781 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg
      841 tttagtgaac cgtcagatca gatctttgtc gatcctacca tccactcgac acacccgcca
      901 gcggccgcca ccatgtctga taatggaccc caaaatcagc gaaatgcacc ccgcattacg
      961 tttggtggac cctcagattc aactggcagt aaccagaatg gagaacgcag tggggcgcga
     1021 tcaaaacaac gtcggcccca aggtttaccc aataatactg cgtcttggtt caccgctctc
     1081 actcaacatg gcaaggaaga ccttaaattc cctcgaggac aaggcgttcc aattaacacc
     1141 aatagcagtc cagatgacca aattggctac taccgaagag ctaccagacg aattcgtggt
     1201 ggtgacggta aaatgaaaga tctcagtcca agatggtatt tctactacct aggaactggg
     1261 ccagaagctg gacttcccta tggtgctaac aaagacggca tcatatgggt tgcaactgag
     1321 ggagccttga atacaccaaa agatcacatt ggcacccgca atcctgctaa caatgctgca
     1381 atcgtgctac aacttcctca aggaacaaca ttgccaaaag gcttctacgc agaagggagc
     1441 agaggcggca gtcaagcctc ttctcgttcc tcatcacgta gtcgcaacag ttcaagaaat
     1501 tcaactccag gcagcagtag gggaacttct cctgctagaa tggctggcaa tggcggtgat
     1561 gctgctcttg ctttgctgct gcttgacaga ttgaaccagc ttgagagcaa aatgtctggt
     1621 aaaggccaac aacaacaagg ccaaactgtc actaagaaat ctgctgctga ggcttctaag
     1681 aagcctcggc aaaaacgtac tgccactaaa gcatacaatg taacacaagc tttcggcaga
     1741 cgtggtccag aacaaaccca aggaaatttt ggggaccagg aactaatcag acaaggaact
     1801 gattacaaac attggccgca aattgcacaa tttgccccca gcgcttcagc gttcttcgga
     1861 atgtcgcgca ttggcatgga agtcacacct tcgggaacgt ggttgaccta cacaggtgcc
     1921 atcaaattgg atgacaaaga tccaaatttc aaagatcaag tcattttgct gaataagcat
     1981 attgacgcat acaaaacatt cccaccaaca gagcctaaaa aggacaaaaa gaagaaggct
     2041 gatgaaactc aagccttacc gcagagacag aagaaacagc aaactgtgac tcttcttcct
     2101 gctgcagatt tggatgattt ctccaaacaa ttgcaacaat ccatgagcag tgctgactca
     2161 actcaggccg aaaaccttta ctttcagtcg ggacggcgaa tcccggggct gataaatccc
     2221 tggaaaagga gatggaagaa gaactttatt gcagtgtctg cggctaaccg gttcaaaaag
     2281 atcagttcaa gcggcgccct agattacgac attcctacga ccgcctccga gaatctctat
     2341 ttccagggtg aatttggctt ggcacaacat gacgaggccg tcgataacaa gtttaataaa
     2401 gaacagcaga acgctttcta tgagatcctt cacttaccaa atctgaacga agagcaacgc
     2461 aacgctttca ttcagagcct gaaagacgat ccctcgcagt ctgcaaatct cctggccgaa
     2521 gccaagaagc tgaatgacgc acaggctcca aaagttgata acaagtttaa caaggagcag
     2581 cagaatgctt tctacgagat attgcacctt cctaatctga atgaggaaca aagaaatgca
     2641 tttatccagt ccctaaaaga cgatccatca cagagtgcca acctgcttgc ggaagccaaa
     2701 aagctcaatg atgcccaggc cccgaaagta gacgcgaacc atcaataagc tagcttgact
     2761 gactgagata cagcgtacct tcagctcaca gacatgataa gatacattga tgagtttgga
     2821 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     2881 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     2941 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac
     3001 aaatgtggta ttggcccatc tctatcggta tcgtagcata accccttggg gcctctaaac
     3061 gggtcttgag gggttttttg tgcccctcgg gccggattgc tatctaccgg cattggcgca
     3121 gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc agattcagca
     3181 acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc agaaatttat
     3241 ccttaaggtc gtcagctatc ctgcaggcga tctctcgatt tcgatcaaga cattccttta
     3301 atggtctttt ctggacacca ctaggggtca gaagtagttc atcaaacttt cttccctccc
     3361 taatctcatt ggttaccttg ggctatcgaa acttaattaa ccagtcaagt cagctacttg
     3421 gcgagatcga cttgtctggg tttcgactac gctcagaatt gcgtcagtca agttcgatct
     3481 ggtccttgct attgcacccg ttctccgatt acgagtttca tttaaatcat gtgagcaaaa
     3541 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     3601 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3661 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     3721 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     3781 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     3841 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3901 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     3961 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     4021 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     4081 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     4141 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     4201 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     4261 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     4321 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     4381 gcgatctgtc tatttcgttc atccatagtt gcatttaaat ttccgaactc tccaaggccc
     4441 tcgtcggaaa atcttcaaac ctttcgtccg atccatcttg caggctacct ctcgaacgaa
     4501 ctatcgcaag tctcttggcc ggccttgcgc cttggctatt gcttggcagc gcctatcgcc
     4561 aggtattact ccaatcccga atatccgaga tcgggatcac ccgagagaag ttcaacctac
     4621 atcctcaatc ccgatctatc cgagatccga ggaatatcga aatcggggcg cgcctggtgt
     4681 accgagaacg atcctctcag tgcgagtctc gacgatccat atcgttgctt ggcagtcagc
     4741 cagtcggaat ccagcttggg acccaggaag tccaatcgtc agatattgta ctcaagcctg
     4801 gtcacggcag cgtaccgatc tgtttaaacc tagatattga tagtctgatc ggtcaacgta
     4861 taatcgagtc ctagcttttg caaacatcta tcaagagaca ggatcagcag gaggctttcg
     4921 catgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     4981 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     5041 gcgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     5101 cgaagaacgc tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     5161 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     5221 ggttgagtat tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     5281 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     5341 tggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     5401 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     5461 gcctgtagca atggcaacaa ccttgcgtaa actattaact ggcgaactac ttactctagc
     5521 ttcccggcaa cagttgatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     5581 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     5641 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     5701 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     5761 ctcactgatt aagcattggt aaccgattct aggtgcattg gcgcagaaaa aaatgcctga
     5821 tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga tacggcttcc
     5881 ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta agatcccgaa
     5941 tcgtttaaac tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta cgcgaacagc
     6001 cgtggcgctc atttgctcgt c
//
