LOCUS       sequence-306089-        7887 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 7887)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7887)
  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-m4134490
FEATURES             Location/Qualifiers
     source          1..7887
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    124..195
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(200..246)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    264..312
                     /note="synthetic polyadenylation signal"
     enhancer        393..696
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..900
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     851..871
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2232..2252
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2256..2321
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    2390..2511
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2401..2420)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2455..2474
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2626..2672
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2702..2838
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2883..2954
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3180..3768)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3260..3279)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4268..5477
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5314..5333
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5481..6509
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    6536..6584
                     /note="synthetic polyadenylation signal"
     CDS             join(6850..6918,6919..7710)
                     /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(7068..7087)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7723..7859
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 gctctatcga atctccgtcg tttcgagctt acgcgaacag ccgtggcgct catttgctcg
       61 tcgggcatcg aatctcgtca gctatcgtca gcttaccttt ttggcagcga tcgcggctcc
      121 cgacatcttg gaccattagc tccacaggta tcttcttccc tctagtggtc ataacagcag
      181 cttcagctac ctctcaattc aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt
      241 atgctatcaa tcgttgcgtt acacacacaa aaaaccaaca cacatccatc ttcgatggat
      301 agcgatttta ttatctaact gctgatcgag tgtagccaga tctagtaatc aattacgggg
      361 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
      421 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
      481 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
      541 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
      601 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
      661 cagtacatct acgtattagt catcgctatt accatgctga tgcggttttg gcagtacatc
      721 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
      781 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
      841 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
      901 ggtttagtga accgtcagat cagatctttg tcgatcctac catccactcg acacacccgc
      961 cagcggccgc caccatgtct gataatggac cacagaacca acgcaatgca ccccgaatca
     1021 cttttggcgg tccgtccgac agtaccggga gcaaccagaa cggcgaaaga tcgggagctc
     1081 gttcaaaaca gcggaggcct caagggttac ccaataacac ggccagctgg ttcacagcgt
     1141 tgacccagca tggcaaggag gacctcaagt tccctcgggg tcagggggtc ccaataaata
     1201 caaacagttc ccctgatgac caaattggat actatagacg cgctactagg aggatccgtg
     1261 gcggggatgg aaaaatgaag gacctttctc ccagatggta cttttattac ctgggtactg
     1321 gcccagaggc cggtctaccg tatggagcaa acaaagatgg cattatttgg gtggccacgg
     1381 aaggagcact gaatacccca aaggatcaca tcggcacccg gaatccagcc aacaatgctg
     1441 ctatcgttct gcaactcccc caggggacaa cattgcctaa aggcttctat gccgagggca
     1501 gtagaggtgg cagccaggcg tcctcacgca gctcttctcg atctcggaac agctcaagaa
     1561 attccactcc cgggagttcc cgaggaacca gcccagcaag gatggctggc aatggtggag
     1621 atgccgctct ggccctactt ctgctggacc gcctcaatca gttagaatca aagatgtcgg
     1681 ggaaggggca gcaacagcag ggacagaccg taactaaaaa gagtgcagcg gaggcctcta
     1741 aaaagcctcg gcagaaacgt acggcaacaa aggcttacaa cgtgacccag gcctttggcc
     1801 gtagaggacc cgaacagaca caagggaact ttggtgacca agagttgata aggcagggaa
     1861 ccgactacaa acactggcct cagatcgccc aattcgcacc gagcgcgagc gcatttttcg
     1921 gaatgtcacg aattggcatg gaggtgacac cttcgggaac ctggctgaca tacactggcg
     1981 ccataaagct cgatgacaaa gatcccaact tcaaggatca agtcatcttg ctgaataagc
     2041 atattgacgc gtataaaact tttccaccca cggaaccgaa gaaggacaag aaaaagaaag
     2101 ctgatgagac acaggcgctt ccacaaaggc agaaaaagca gcagacggtt actctactgc
     2161 ccgccgcaga cctagatgac ttctccaaac agcttcagca gtctatgagt agcgccgact
     2221 caacccaagc tgaaaacctt tactttcagt cgatggatta taaggaccac gatggagact
     2281 acaaagatca tgacattgat tacaaggacg acgatgacaa ataaggatcc gcaggcctct
     2341 gctagcttga ctgactgaga tacagcgtac cttcagctca cagacatgat aagatacatt
     2401 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     2461 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     2521 aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag
     2581 taaaacctct acaaatgtgg tattggccca tctctatcgg tatcgtagca taaccccttg
     2641 gggcctctaa acgggtcttg aggggttttt tgtgcccctc gggccggatt gctatctacc
     2701 ggcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact cccacatatg
     2761 ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt gtcctcctta
     2821 ccagaaattt atccttaagg tcgtcagcta tcctgcaggc gatctctcga tttcgatcaa
     2881 gacattcctt taatggtctt ttctggacac cactaggggt cagaagtagt tcatcaaact
     2941 ttcttccctc cctaatctca ttggttacct tgggctatcg aaacttaatt aaccagtcaa
     3001 gtcagctact tggcgagatc gacttgtctg ggtttcgact acgctcagaa ttgcgtcagt
     3061 caagttcgat ctggtccttg ctattgcacc cgttctccga ttacgagttt catttaaatc
     3121 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     3181 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     3241 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     3301 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     3361 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     3421 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     3481 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     3541 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     3601 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3661 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     3721 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3781 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     3841 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     3901 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     3961 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcatttaa atttccgaac
     4021 tctccaaggc cctcgtcgga aaatcttcaa acctttcgtc cgatccatct tgcaggctac
     4081 ctctcgaacg aactatcgca agtctcttgg ccggccttgc gccttggcta ttgcttggca
     4141 gcgcctatcg ccaggtatta ctccaatccc gaatatccga gatcgggatc acccgagaga
     4201 agttcaacct acatcctcaa tcccgatcta tccgagatcc gaggaatatc gaaatcgggg
     4261 cgcgcctggc ctccgcgccg ggttttggcg cctcccgcgg gcgcccccct cgtcacggcg
     4321 agcgctgcca cgtcagacga agggcgcagg agcgtcctga tccttccgcc cggacgctca
     4381 ggacagcggc ccgctgctca taagactcgg ccttagaacc ccagtatcag cagaaggaca
     4441 ttttaggacg ggacttgggt gactctaggg cactggtttt ctttccagag agcggaacag
     4501 gcgaggaaaa gtagtccctt ctcggcgatt ctgcggaggg atctccgtgg ggcggtgaac
     4561 gccgatgatt atataaggac gcgccgggtg tggcacagct agttccgtcg cagccgggat
     4621 ttgggtcgcg gttcttgttt gtggatcgct gtgatcgtca cttggtgagt agcgggctgc
     4681 tgggctggcc ggggctttcg tggccgccgg gccgctcggt gggacggaag cgtgtggaga
     4741 gaccgccaag ggctgtagtc tgggtccgcg agcaaggttg ccctgaactg ggggttgggg
     4801 ggagcgcagc aaaatggcgg ctgttcccga gtcttgaatg gaagacgctt gtgaggcggg
     4861 ctgtgaggtc gttgaaacaa ggtggggggc atggtgggcg gcaagaaccc aaggtcttga
     4921 gcccttcgct aatgcgggaa agctcttatt cgggtgagat gggctgggca ccatctgggg
     4981 accctgacgt gaagtttgtc actgactgga gaactcggtt tgtcgtctgt tgcgggggcg
     5041 gcagttatgg cggtgccgtt gggcagtgca cccgtacctt tgggagcgcg cgccctcgtc
     5101 gtgtcgtgac gtcacccgtt ctgttggctt ataatgcagg gtggggccac ctgccggtag
     5161 gtgtgcggta ggcttttctc cgtcgcagga cgcagggttc gggcctaggg taggctctcc
     5221 tgaatcgaca ggcgccggac ctctggtgag gggagggata agtgaggcgt cagtttcttt
     5281 ggtcggtttt atgtacctat cttcttaagt agctgaagct ccggttttga actatgcgct
     5341 cggggttggc gagtgtgttt tgtgaagttt tttaggcacc ttttgaaatg taatcatttg
     5401 ggtcaatatg taattttcag tgttagactt gtaaattgtc cgctaaattc tggccgtttt
     5461 tggctttttt gttagacaac atgggtaaaa agcctgaact caccgcgacg tctgtcgaga
     5521 agtttctgat cgaaaagttc gacagcgtct ccgacctgat gcagctctcg gagggcgaag
     5581 aatctcgtgc tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct
     5641 gcgccgatgg tttctacaaa gatcgttatg tttatcggca ctttgcatcc gccgcgctcc
     5701 cgattccgga agtgcttgac attggggagt tcagcgagag cctgacctat tgcatctccc
     5761 gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc
     5821 agccggtcgc ggaggcaatg gatgccatcg ctgccgccga tcttagccag acgagcgggt
     5881 tcggcccatt cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg
     5941 cgattgctga tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt
     6001 ccgtcgcgca ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc
     6061 acctcgtgca cgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag
     6121 cggtcattga ctggagcgag gcgatgttcg gggattccca atacgaggtc gccaacatct
     6181 tcttctggag gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc
     6241 atccggagct tgcaggatcg ccccggctcc gggcgtatat gctccgcatt ggtcttgacc
     6301 aactctatca gagcttggtt gacggcaatt tcgatgatgc agcttgggcg cagggtcgat
     6361 gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa
     6421 gcgccgccgt ctggaccgat ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc
     6481 ccagcactcg tccgagggca aaggaataag ctagtatgta agcctagtct tagataataa
     6541 aatcgctatc catcgaagat ggatgtgtgt tggttttttg tgtgtgtaac gctaggcgcg
     6601 cctggtgtac cgagaacgat cctctcagtg cgagtctcga cgatccatat cgttgcttgg
     6661 cagtcagcca gtcggaatcc agcttgggac ccaggaagtc caatcgtcag atattgtact
     6721 caagcctggt cacggcagcg taccgatctg tttaaaccta gatattgata gtctgatcgg
     6781 tcaacgtata atcgagtcct agcttttgca aacatctatc aagagacagg atcagcagga
     6841 ggctttcgca tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     6901 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     6961 ttgggtgcgc gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     7021 tttcgccccg aagaacgctt tccaatgatg agcactttta aagttctgct atgtggcgcg
     7081 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     7141 aatgacttgg ttgagtattc accagtcaca gaaaagcatc ttacggatgg catgacagta
     7201 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     7261 acaacgattg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     7321 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     7381 accacgatgc ctgtagcaat ggcaacaacc ttgcgtaaac tattaactgg cgaactactt
     7441 actctagctt cccggcaaca gttgatagac tggatggagg cggataaagt tgcaggacca
     7501 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     7561 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     7621 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     7681 ataggtgcct cactgattaa gcattggtaa ccgattctag gtgcattggc gcagaaaaaa
     7741 atgcctgatg cgacgctgcg cgtcttatac tcccacatat gccagattca gcaacggata
     7801 cggcttcccc aacttgccca cttccatacg tgtcctcctt accagaaatt tatccttaag
     7861 atcccgaatc gtttaaactc gactctg
//
