LOCUS       sequence-300229-        7845 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 7845)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7845)
  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-m4134336
FEATURES             Location/Qualifiers
     source          1..7845
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        100..403
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        405..607
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     558..578
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1939..1959
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1960..1986
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2055..2176
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2066..2085)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2120..2139
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2291..2337
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2367..2503
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2548..2619
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2845..3433)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2925..2944)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3933..5142
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4979..4998
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5146..6174
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    6201..6249
                     /note="synthetic polyadenylation signal"
     CDS             join(6515..6583,6584..7375)
                     /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(6733..6752)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7388..7524
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7676..7747
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7752..7798)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    join(7816..7845,1..19)
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 gatggatagc gattttatta tctaactgct gatcgagtgt agccagatct agtaatcaat
       61 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
      121 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
      181 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
      241 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
      301 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
      361 tacttggcag tacatctacg tattagtcat cgctattacc atgctgatgc ggttttggca
      421 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
      481 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
      541 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
      601 cagagctggt ttagtgaacc gtcagatcag atctttgtcg atcctaccat ccactcgaca
      661 cacccgccag cggccgccac catgtctgat aatggacccc aaaatcagcg aaatgcaccc
      721 cgcattacgt ttggtggacc ctcagattca actggcagta accagaatgg agaacgcagt
      781 ggggcgcgat caaaacaacg tcggccccaa ggtttaccca ataatactgc gtcttggttc
      841 accgctctca ctcaacatgg caaggaagac cttaaattcc ctcgaggaca aggcgttcca
      901 attaacacca atagcagtcc agatgaccaa attggctact accgaagagc taccagacga
      961 attcgtggtg gtgacggtaa aatgaaagat ctcagtccaa gatggtattt ctactaccta
     1021 ggaactgggc cagaagctgg acttccctat ggtgctaaca aagacggcat catatgggtt
     1081 gcaactgagg gagccttgaa tacaccaaaa gatcacattg gcacccgcaa tcctgctaac
     1141 aatgctgcaa tcgtgctaca acttcctcaa ggaacaacat tgccaaaagg cttctacgca
     1201 gaagggagca gaggcggcag tcaagcctct tctcgttcct catcacgtag tcgcaacagt
     1261 tcaagaaatt caactccagg cagcagtagg ggaacttctc ctgctagaat ggctggcaat
     1321 ggcggtgatg ctgctcttgc tttgctgctg cttgacagat tgaaccagct tgagagcaaa
     1381 atgtctggta aaggccaaca acaacaaggc caaactgtca ctaagaaatc tgctgctgag
     1441 gcttctaaga agcctcggca aaaacgtact gccactaaag catacaatgt aacacaagct
     1501 ttcggcagac gtggtccaga acaaacccaa ggaaattttg gggaccagga actaatcaga
     1561 caaggaactg attacaaaca ttggccgcaa attgcacaat ttgcccccag cgcttcagcg
     1621 ttcttcggaa tgtcgcgcat tggcatggaa gtcacacctt cgggaacgtg gttgacctac
     1681 acaggtgcca tcaaattgga tgacaaagat ccaaatttca aagatcaagt cattttgctg
     1741 aataagcata ttgacgcata caaaacattc ccaccaacag agcctaaaaa ggacaaaaag
     1801 aagaaggctg atgaaactca agccttaccg cagagacaga agaaacagca aactgtgact
     1861 cttcttcctg ctgcagattt ggatgatttc tccaaacaat tgcaacaatc catgagcagt
     1921 gctgactcaa ctcaggccga aaacctttac tttcagtcgt acccctatga cgtaccagat
     1981 tatgcgtaag gatccgcagg cctctgctag cttgactgac tgagatacag cgtaccttca
     2041 gctcacagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     2101 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     2161 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     2221 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtattg gcccatctct
     2281 atcggtatcg tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgtgc
     2341 ccctcgggcc ggattgctat ctaccggcat tggcgcagaa aaaaatgcct gatgcgacgc
     2401 tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg
     2461 cccacttcca tacgtgtcct ccttaccaga aatttatcct taaggtcgtc agctatcctg
     2521 caggcgatct ctcgatttcg atcaagacat tcctttaatg gtcttttctg gacaccacta
     2581 ggggtcagaa gtagttcatc aaactttctt ccctccctaa tctcattggt taccttgggc
     2641 tatcgaaact taattaacca gtcaagtcag ctacttggcg agatcgactt gtctgggttt
     2701 cgactacgct cagaattgcg tcagtcaagt tcgatctggt ccttgctatt gcacccgttc
     2761 tccgattacg agtttcattt aaatcatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     2821 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     2881 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     2941 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     3001 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     3061 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     3121 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     3181 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     3241 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     3301 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     3361 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     3421 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     3481 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     3541 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     3601 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     3661 catagttgca tttaaatttc cgaactctcc aaggccctcg tcggaaaatc ttcaaacctt
     3721 tcgtccgatc catcttgcag gctacctctc gaacgaacta tcgcaagtct cttggccggc
     3781 cttgcgcctt ggctattgct tggcagcgcc tatcgccagg tattactcca atcccgaata
     3841 tccgagatcg ggatcacccg agagaagttc aacctacatc ctcaatcccg atctatccga
     3901 gatccgagga atatcgaaat cggggcgcgc ctggcctccg cgccgggttt tggcgcctcc
     3961 cgcgggcgcc cccctcgtca cggcgagcgc tgccacgtca gacgaagggc gcaggagcgt
     4021 cctgatcctt ccgcccggac gctcaggaca gcggcccgct gctcataaga ctcggcctta
     4081 gaaccccagt atcagcagaa ggacatttta ggacgggact tgggtgactc tagggcactg
     4141 gttttctttc cagagagcgg aacaggcgag gaaaagtagt cccttctcgg cgattctgcg
     4201 gagggatctc cgtggggcgg tgaacgccga tgattatata aggacgcgcc gggtgtggca
     4261 cagctagttc cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga tcgctgtgat
     4321 cgtcacttgg tgagtagcgg gctgctgggc tggccggggc tttcgtggcc gccgggccgc
     4381 tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt ccgcgagcaa
     4441 ggttgccctg aactgggggt tggggggagc gcagcaaaat ggcggctgtt cccgagtctt
     4501 gaatggaaga cgcttgtgag gcgggctgtg aggtcgttga aacaaggtgg ggggcatggt
     4561 gggcggcaag aacccaaggt cttgagccct tcgctaatgc gggaaagctc ttattcgggt
     4621 gagatgggct gggcaccatc tggggaccct gacgtgaagt ttgtcactga ctggagaact
     4681 cggtttgtcg tctgttgcgg gggcggcagt tatggcggtg ccgttgggca gtgcacccgt
     4741 acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac ccgttctgtt ggcttataat
     4801 gcagggtggg gccacctgcc ggtaggtgtg cggtaggctt ttctccgtcg caggacgcag
     4861 ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc cggacctctg gtgaggggag
     4921 ggataagtga ggcgtcagtt tctttggtcg gttttatgta cctatcttct taagtagctg
     4981 aagctccggt tttgaactat gcgctcgggg ttggcgagtg tgttttgtga agttttttag
     5041 gcaccttttg aaatgtaatc atttgggtca atatgtaatt ttcagtgtta gacttgtaaa
     5101 ttgtccgcta aattctggcc gtttttggct tttttgttag acaacatggg taaaaagcct
     5161 gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag cgtctccgac
     5221 ctgatgcagc tctcggaggg cgaagaatct cgtgctttca gcttcgatgt aggagggcgt
     5281 ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg ttatgtttat
     5341 cggcactttg catccgccgc gctcccgatt ccggaagtgc ttgacattgg ggagttcagc
     5401 gagagcctga cctattgcat ctcccgccgt gcacagggtg tcacgttgca agacctgcct
     5461 gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg caatggatgc catcgctgcc
     5521 gccgatctta gccagacgag cgggttcggc ccattcggac cgcaaggaat cggtcaatac
     5581 actacatggc gtgatttcat atgcgcgatt gctgatcccc atgtgtatca ctggcaaact
     5641 gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg
     5701 gccgaggact gccccgaagt ccggcacctc gtgcacgcgg atttcggctc caacaatgtc
     5761 ctgacggaca atggccgcat aacagcggtc attgactgga gcgaggcgat gttcggggat
     5821 tcccaatacg aggtcgccaa catcttcttc tggaggccgt ggttggcttg tatggagcag
     5881 cagacgcgct acttcgagcg gaggcatccg gagcttgcag gatcgccccg gctccgggcg
     5941 tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg caatttcgat
     6001 gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc
     6061 gggcgtacac aaatcgcccg cagaagcgcc gccgtctgga ccgatggctg tgtagaagta
     6121 ctcgccgata gtggaaaccg acgccccagc actcgtccga gggcaaagga ataagctagt
     6181 atgtaagcct agtcttagat aataaaatcg ctatccatcg aagatggatg tgtgttggtt
     6241 ttttgtgtgt gtaacgctag gcgcgcctgg tgtaccgaga acgatcctct cagtgcgagt
     6301 ctcgacgatc catatcgttg cttggcagtc agccagtcgg aatccagctt gggacccagg
     6361 aagtccaatc gtcagatatt gtactcaagc ctggtcacgg cagcgtaccg atctgtttaa
     6421 acctagatat tgatagtctg atcggtcaac gtataatcga gtcctagctt ttgcaaacat
     6481 ctatcaagag acaggatcag caggaggctt tcgcatgagt attcaacatt tccgtgtcgc
     6541 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     6601 gaaagtaaaa gatgctgaag atcagttggg tgcgcgagtg ggttacatcg aactggatct
     6661 caacagcggt aagatccttg agagttttcg ccccgaagaa cgctttccaa tgatgagcac
     6721 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     6781 cggtcgccgc atacactatt ctcagaatga cttggttgag tattcaccag tcacagaaaa
     6841 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     6901 taacactgcg gccaacttac ttctgacaac gattggagga ccgaaggagc taaccgcttt
     6961 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     7021 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caaccttgcg
     7081 taaactatta actggcgaac tacttactct agcttcccgg caacagttga tagactggat
     7141 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     7201 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     7261 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     7321 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaaccgat
     7381 tctaggtgca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca
     7441 catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc atacgtgtcc
     7501 tccttaccag aaatttatcc ttaagatccc gaatcgttta aactcgactc tggctctatc
     7561 gaatctccgt cgtttcgagc ttacgcgaac agccgtggcg ctcatttgct cgtcgggcat
     7621 cgaatctcgt cagctatcgt cagcttacct ttttggcagc gatcgcggct cccgacatct
     7681 tggaccatta gctccacagg tatcttcttc cctctagtgg tcataacagc agcttcagct
     7741 acctctcaat tcaaaaaacc cctcaagacc cgtttagagg ccccaagggg ttatgctatc
     7801 aatcgttgcg ttacacacac aaaaaaccaa cacacatcca tcttc
//
