LOCUS       sequence-307247-        5535 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 5535)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5535)
  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-m4133772
FEATURES             Location/Qualifiers
     source          1..5535
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      242..378
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    530..601
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(606..652)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    670..718
                     /note="synthetic polyadenylation signal"
     enhancer        799..1102
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1104..1306
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1257..1277
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2638..2658
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2662..2727
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    2780..2901
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2791..2810)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2845..2864
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      3016..3062
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      3092..3228
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3273..3344
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3570..4158)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3650..3669)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(4904..4972,4973..5535,1..229)
                     /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(5122..5141)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
       61 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
      121 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
      181 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac cgattctagg
      241 tgcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact cccacatatg
      301 ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt gtcctcctta
      361 ccagaaattt atccttaaga tcccgaatcg tttaaactcg actctggctc tatcgaatct
      421 ccgtcgtttc gagcttacgc gaacagccgt ggcgctcatt tgctcgtcgg gcatcgaatc
      481 tcgtcagcta tcgtcagctt acctttttgg cagcgatcgc ggctcccgac atcttggacc
      541 attagctcca caggtatctt cttccctcta gtggtcataa cagcagcttc agctacctct
      601 caattcaaaa aacccctcaa gacccgttta gaggccccaa ggggttatgc tatcaatcgt
      661 tgcgttacac acacaaaaaa ccaacacaca tccatcttcg atggatagcg attttattat
      721 ctaactgctg atcgagtgta gccagatcta gtaatcaatt acggggtcat tagttcatag
      781 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
      841 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
      901 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
      961 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     1021 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     1081 attagtcatc gctattacca tgctgatgcg gttttggcag tacatcaatg ggcgtggata
     1141 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     1201 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
     1261 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg
     1321 tcagatcaga tctttgtcga tcctaccatc cactcgacac acccgccagc ggccgccacc
     1381 atgtctgata atggacccca aaatcagcga aatgcacccc gcattacgtt tggtggaccc
     1441 tcagattcaa ctggcagtaa ccagaatgga gaacgcagtg gggcgcgatc aaaacaacgt
     1501 cggccccaag gtttacccaa taatactgcg tcttggttca ccgctctcac tcaacatggc
     1561 aaggaagacc ttaaattccc tcgaggacaa ggcgttccaa ttaacaccaa tagcagtcca
     1621 gatgaccaaa ttggctacta ccgaagagct accagacgaa ttcgtggtgg tgacggtaaa
     1681 atgaaagatc tcagtccaag atggtatttc tactacctag gaactgggcc agaagctgga
     1741 cttccctatg gtgctaacaa agacggcatc atatgggttg caactgaggg agccttgaat
     1801 acaccaaaag atcacattgg cacccgcaat cctgctaaca atgctgcaat cgtgctacaa
     1861 cttcctcaag gaacaacatt gccaaaaggc ttctacgcag aagggagcag aggcggcagt
     1921 caagcctctt ctcgttcctc atcacgtagt cgcaacagtt caagaaattc aactccaggc
     1981 agcagtaggg gaacttctcc tgctagaatg gctggcaatg gcggtgatgc tgctcttgct
     2041 ttgctgctgc ttgacagatt gaaccagctt gagagcaaaa tgtctggtaa aggccaacaa
     2101 caacaaggcc aaactgtcac taagaaatct gctgctgagg cttctaagaa gcctcggcaa
     2161 aaacgtactg ccactaaagc atacaatgta acacaagctt tcggcagacg tggtccagaa
     2221 caaacccaag gaaattttgg ggaccaggaa ctaatcagac aaggaactga ttacaaacat
     2281 tggccgcaaa ttgcacaatt tgcccccagc gcttcagcgt tcttcggaat gtcgcgcatt
     2341 ggcatggaag tcacaccttc gggaacgtgg ttgacctaca caggtgccat caaattggat
     2401 gacaaagatc caaatttcaa agatcaagtc attttgctga ataagcatat tgacgcatac
     2461 aaaacattcc caccaacaga gcctaaaaag gacaaaaaga agaaggctga tgaaactcaa
     2521 gccttaccgc agagacagaa gaaacagcaa actgtgactc ttcttcctgc tgcagatttg
     2581 gatgatttct ccaaacaatt gcaacaatcc atgagcagtg ctgactcaac tcaggccgaa
     2641 aacctttact ttcagtcgat ggattataag gaccacgatg gagactacaa agatcatgac
     2701 attgattaca aggacgacga tgacaaataa gctagcttga ctgactgaga tacagcgtac
     2761 cttcagctca cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     2821 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     2881 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     2941 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tattggccca
     3001 tctctatcgg tatcgtagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     3061 tgtgcccctc gggccggatt gctatctacc ggcattggcg cagaaaaaaa tgcctgatgc
     3121 gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca
     3181 acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaagg tcgtcagcta
     3241 tcctgcaggc gatctctcga tttcgatcaa gacattcctt taatggtctt ttctggacac
     3301 cactaggggt cagaagtagt tcatcaaact ttcttccctc cctaatctca ttggttacct
     3361 tgggctatcg aaacttaatt aaccagtcaa gtcagctact tggcgagatc gacttgtctg
     3421 ggtttcgact acgctcagaa ttgcgtcagt caagttcgat ctggtccttg ctattgcacc
     3481 cgttctccga ttacgagttt catttaaatc atgtgagcaa aaggccagca aaaggccagg
     3541 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3601 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3661 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     3721 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     3781 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     3841 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     3901 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     3961 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     4021 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     4081 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     4141 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     4201 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     4261 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     4321 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     4381 tcatccatag ttgcatttaa atttccgaac tctccaaggc cctcgtcgga aaatcttcaa
     4441 acctttcgtc cgatccatct tgcaggctac ctctcgaacg aactatcgca agtctcttgg
     4501 ccggccttgc gccttggcta ttgcttggca gcgcctatcg ccaggtatta ctccaatccc
     4561 gaatatccga gatcgggatc acccgagaga agttcaacct acatcctcaa tcccgatcta
     4621 tccgagatcc gaggaatatc gaaatcgggg cgcgcctggt gtaccgagaa cgatcctctc
     4681 agtgcgagtc tcgacgatcc atatcgttgc ttggcagtca gccagtcgga atccagcttg
     4741 ggacccagga agtccaatcg tcagatattg tactcaagcc tggtcacggc agcgtaccga
     4801 tctgtttaaa cctagatatt gatagtctga tcggtcaacg tataatcgag tcctagcttt
     4861 tgcaaacatc tatcaagaga caggatcagc aggaggcttt cgcatgagta ttcaacattt
     4921 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     4981 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcgcgagtgg gttacatcga
     5041 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gctttccaat
     5101 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     5161 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt attcaccagt
     5221 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     5281 catgagtgat aacactgcgg ccaacttact tctgacaacg attggaggac cgaaggagct
     5341 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     5401 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     5461 aaccttgcgt aaactattaa ctggcgaact acttactcta gcttcccggc aacagttgat
     5521 agactggatg gaggc
//
