LOCUS       sequence-306183-        7451 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 7451)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7451)
  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-m4134538
FEATURES             Location/Qualifiers
     source          1..7451
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    109..180
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(185..231)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    249..297
                     /note="synthetic polyadenylation signal"
     enhancer        378..681
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        683..885
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     836..856
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2217..2237
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2238..2315
                     /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    2368..2489
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2379..2398)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2433..2452
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2604..2650
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2680..2816
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2861..2932
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3158..3746)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3238..3257)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4246..5455
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5292..5311
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5459..6058
                     /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(5459..5478)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5955..5975
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    6085..6133
                     /note="synthetic polyadenylation signal"
     CDS             join(6399..6467,6468..7259)
                     /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(6617..6636)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7272..7408
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt gctcgtcggg catcgaatct
       61 cgtcagctat cgtcagctta cctttttggc agcgatcgcg gctcccgaca tcttggacca
      121 ttagctccac aggtatcttc ttccctctag tggtcataac agcagcttca gctacctctc
      181 aattcaaaaa acccctcaag acccgtttag aggccccaag gggttatgct atcaatcgtt
      241 gcgttacaca cacaaaaaac caacacacat ccatcttcga tggatagcga ttttattatc
      301 taactgctga tcgagtgtag ccagatctag taatcaatta cggggtcatt agttcatagc
      361 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
      421 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
      481 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
      541 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
      601 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
      661 ttagtcatcg ctattaccat gctgatgcgg ttttggcagt acatcaatgg gcgtggatag
      721 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
      781 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
      841 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt
      901 cagatcagat ctttgtcgat cctaccatcc actcgacaca cccgccagcg gccgccacca
      961 tgtctgataa tggaccacag aaccaacgca atgcaccccg aatcactttt ggcggtccgt
     1021 ccgacagtac cgggagcaac cagaacggcg aaagatcggg agctcgttca aaacagcgga
     1081 ggcctcaagg gttacccaat aacacggcca gctggttcac agcgttgacc cagcatggca
     1141 aggaggacct caagttccct cggggtcagg gggtcccaat aaatacaaac agttcccctg
     1201 atgaccaaat tggatactat agacgcgcta ctaggaggat ccgtggcggg gatggaaaaa
     1261 tgaaggacct ttctcccaga tggtactttt attacctggg tactggccca gaggccggtc
     1321 taccgtatgg agcaaacaaa gatggcatta tttgggtggc cacggaagga gcactgaata
     1381 ccccaaagga tcacatcggc acccggaatc cagccaacaa tgctgctatc gttctgcaac
     1441 tcccccaggg gacaacattg cctaaaggct tctatgccga gggcagtaga ggtggcagcc
     1501 aggcgtcctc acgcagctct tctcgatctc ggaacagctc aagaaattcc actcccggga
     1561 gttcccgagg aaccagccca gcaaggatgg ctggcaatgg tggagatgcc gctctggccc
     1621 tacttctgct ggaccgcctc aatcagttag aatcaaagat gtcggggaag gggcagcaac
     1681 agcagggaca gaccgtaact aaaaagagtg cagcggaggc ctctaaaaag cctcggcaga
     1741 aacgtacggc aacaaaggct tacaacgtga cccaggcctt tggccgtaga ggacccgaac
     1801 agacacaagg gaactttggt gaccaagagt tgataaggca gggaaccgac tacaaacact
     1861 ggcctcagat cgcccaattc gcaccgagcg cgagcgcatt tttcggaatg tcacgaattg
     1921 gcatggaggt gacaccttcg ggaacctggc tgacatacac tggcgccata aagctcgatg
     1981 acaaagatcc caacttcaag gatcaagtca tcttgctgaa taagcatatt gacgcgtata
     2041 aaacttttcc acccacggaa ccgaagaagg acaagaaaaa gaaagctgat gagacacagg
     2101 cgcttccaca aaggcagaaa aagcagcaga cggttactct actgcccgcc gcagacctag
     2161 atgacttctc caaacagctt cagcagtcta tgagtagcgc cgactcaacc caagctgaaa
     2221 acctttactt tcagtcgaag aggcgatgga aaaagaattt tatcgcggtg agtgcagcca
     2281 acagattcaa aaagatatcc tcgtctggtg ccctctaagc tagcttgact gactgagata
     2341 cagcgtacct tcagctcaca gacatgataa gatacattga tgagtttgga caaaccacaa
     2401 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     2461 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     2521 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     2581 ttggcccatc tctatcggta tcgtagcata accccttggg gcctctaaac gggtcttgag
     2641 gggttttttg tgcccctcgg gccggattgc tatctaccgg cattggcgca gaaaaaaatg
     2701 cctgatgcga cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg
     2761 cttccccaac ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaaggtc
     2821 gtcagctatc ctgcaggcga tctctcgatt tcgatcaaga cattccttta atggtctttt
     2881 ctggacacca ctaggggtca gaagtagttc atcaaacttt cttccctccc taatctcatt
     2941 ggttaccttg ggctatcgaa acttaattaa ccagtcaagt cagctacttg gcgagatcga
     3001 cttgtctggg tttcgactac gctcagaatt gcgtcagtca agttcgatct ggtccttgct
     3061 attgcacccg ttctccgatt acgagtttca tttaaatcat gtgagcaaaa ggccagcaaa
     3121 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     3181 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     3241 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3301 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     3361 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3421 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     3481 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     3541 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     3601 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     3661 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     3721 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     3781 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     3841 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     3901 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     3961 tatttcgttc atccatagtt gcatttaaat ttccgaactc tccaaggccc tcgtcggaaa
     4021 atcttcaaac ctttcgtccg atccatcttg caggctacct ctcgaacgaa ctatcgcaag
     4081 tctcttggcc ggccttgcgc cttggctatt gcttggcagc gcctatcgcc aggtattact
     4141 ccaatcccga atatccgaga tcgggatcac ccgagagaag ttcaacctac atcctcaatc
     4201 ccgatctatc cgagatccga ggaatatcga aatcggggcg cgcctggcct ccgcgccggg
     4261 ttttggcgcc tcccgcgggc gcccccctcg tcacggcgag cgctgccacg tcagacgaag
     4321 ggcgcaggag cgtcctgatc cttccgcccg gacgctcagg acagcggccc gctgctcata
     4381 agactcggcc ttagaacccc agtatcagca gaaggacatt ttaggacggg acttgggtga
     4441 ctctagggca ctggttttct ttccagagag cggaacaggc gaggaaaagt agtcccttct
     4501 cggcgattct gcggagggat ctccgtgggg cggtgaacgc cgatgattat ataaggacgc
     4561 gccgggtgtg gcacagctag ttccgtcgca gccgggattt gggtcgcggt tcttgtttgt
     4621 ggatcgctgt gatcgtcact tggtgagtag cgggctgctg ggctggccgg ggctttcgtg
     4681 gccgccgggc cgctcggtgg gacggaagcg tgtggagaga ccgccaaggg ctgtagtctg
     4741 ggtccgcgag caaggttgcc ctgaactggg ggttgggggg agcgcagcaa aatggcggct
     4801 gttcccgagt cttgaatgga agacgcttgt gaggcgggct gtgaggtcgt tgaaacaagg
     4861 tggggggcat ggtgggcggc aagaacccaa ggtcttgagc ccttcgctaa tgcgggaaag
     4921 ctcttattcg ggtgagatgg gctgggcacc atctggggac cctgacgtga agtttgtcac
     4981 tgactggaga actcggtttg tcgtctgttg cgggggcggc agttatggcg gtgccgttgg
     5041 gcagtgcacc cgtacctttg ggagcgcgcg ccctcgtcgt gtcgtgacgt cacccgttct
     5101 gttggcttat aatgcagggt ggggccacct gccggtaggt gtgcggtagg cttttctccg
     5161 tcgcaggacg cagggttcgg gcctagggta ggctctcctg aatcgacagg cgccggacct
     5221 ctggtgaggg gagggataag tgaggcgtca gtttctttgg tcggttttat gtacctatct
     5281 tcttaagtag ctgaagctcc ggttttgaac tatgcgctcg gggttggcga gtgtgttttg
     5341 tgaagttttt taggcacctt ttgaaatgta atcatttggg tcaatatgta attttcagtg
     5401 ttagacttgt aaattgtccg ctaaattctg gccgtttttg gcttttttgt tagacaacat
     5461 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg
     5521 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg
     5581 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat
     5641 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag
     5701 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc
     5761 ccggctggcc gcgcagcaac agatggaggg cctcctggcg ccgcaccggc ccaaggagcc
     5821 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag
     5881 cgccgtcgtg ctccccggag tggaggccgc cgagcgcgcc ggggtgcccg ccttcctgga
     5941 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga
     6001 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctaggc
     6061 tagtatgtaa gcctagtctt agataataaa atcgctatcc atcgaagatg gatgtgtgtt
     6121 ggttttttgt gtgtgtaacg ctaggcgcgc ctggtgtacc gagaacgatc ctctcagtgc
     6181 gagtctcgac gatccatatc gttgcttggc agtcagccag tcggaatcca gcttgggacc
     6241 caggaagtcc aatcgtcaga tattgtactc aagcctggtc acggcagcgt accgatctgt
     6301 ttaaacctag atattgatag tctgatcggt caacgtataa tcgagtccta gcttttgcaa
     6361 acatctatca agagacagga tcagcaggag gctttcgcat gagtattcaa catttccgtg
     6421 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     6481 tggtgaaagt aaaagatgct gaagatcagt tgggtgcgcg agtgggttac atcgaactgg
     6541 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgcttt ccaatgatga
     6601 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     6661 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtattca ccagtcacag
     6721 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     6781 gtgataacac tgcggccaac ttacttctga caacgattgg aggaccgaag gagctaaccg
     6841 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     6901 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacct
     6961 tgcgtaaact attaactggc gaactactta ctctagcttc ccggcaacag ttgatagact
     7021 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     7081 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     7141 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     7201 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     7261 cgattctagg tgcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact
     7321 cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt
     7381 gtcctcctta ccagaaattt atccttaaga tcccgaatcg tttaaactcg actctggctc
     7441 tatcgaatct c
//
