LOCUS       sequence-307391-        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-m4134533
FEATURES             Location/Qualifiers
     source          1..7845
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     259..278
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             426..1454
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    1481..1529
                     /note="synthetic polyadenylation signal"
     CDS             join(1795..1863,1864..2655)
                     /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(2013..2032)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      2668..2804
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2956..3027
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3032..3078)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3096..3144
                     /note="synthetic polyadenylation signal"
     enhancer        3225..3528
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3530..3732
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3683..3703
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             5064..5084
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             5085..5111
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    5180..5301
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5191..5210)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5245..5264
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      5416..5462
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      5492..5628
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5673..5744
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(5970..6558)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6050..6069)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(7058..7845,1..422)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
ORIGIN
        1 ccgttgggca gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac
       61 ccgttctgtt ggcttataat gcagggtggg gccacctgcc ggtaggtgtg cggtaggctt
      121 ttctccgtcg caggacgcag ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc
      181 cggacctctg gtgaggggag ggataagtga ggcgtcagtt tctttggtcg gttttatgta
      241 cctatcttct taagtagctg aagctccggt tttgaactat gcgctcgggg ttggcgagtg
      301 tgttttgtga agttttttag gcaccttttg aaatgtaatc atttgggtca atatgtaatt
      361 ttcagtgtta gacttgtaaa ttgtccgcta aattctggcc gtttttggct tttttgttag
      421 acaacatggg taaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa
      481 agttcgacag cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca
      541 gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct
      601 acaaagatcg ttatgtttat cggcactttg catccgccgc gctcccgatt ccggaagtgc
      661 ttgacattgg ggagttcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg
      721 tcacgttgca agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg
      781 caatggatgc catcgctgcc gccgatctta gccagacgag cgggttcggc ccattcggac
      841 cgcaaggaat cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc
      901 atgtgtatca ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc
      961 tcgatgagct gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg
     1021 atttcggctc caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga
     1081 gcgaggcgat gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt
     1141 ggttggcttg tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag
     1201 gatcgccccg gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct
     1261 tggttgacgg caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc
     1321 gatccggagc cgggactgtc gggcgtacac aaatcgcccg cagaagcgcc gccgtctgga
     1381 ccgatggctg tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga
     1441 gggcaaagga ataagctagt atgtaagcct agtcttagat aataaaatcg ctatccatcg
     1501 aagatggatg tgtgttggtt ttttgtgtgt gtaacgctag gcgcgcctgg tgtaccgaga
     1561 acgatcctct cagtgcgagt ctcgacgatc catatcgttg cttggcagtc agccagtcgg
     1621 aatccagctt gggacccagg aagtccaatc gtcagatatt gtactcaagc ctggtcacgg
     1681 cagcgtaccg atctgtttaa acctagatat tgatagtctg atcggtcaac gtataatcga
     1741 gtcctagctt ttgcaaacat ctatcaagag acaggatcag caggaggctt tcgcatgagt
     1801 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     1861 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcgcgagtg
     1921 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     1981 cgctttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     2041 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     2101 tattcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     2161 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gattggagga
     2221 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     2281 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     2341 gcaatggcaa caaccttgcg taaactatta actggcgaac tacttactct agcttcccgg
     2401 caacagttga tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     2461 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     2521 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     2581 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     2641 attaagcatt ggtaaccgat tctaggtgca ttggcgcaga aaaaaatgcc tgatgcgacg
     2701 ctgcgcgtct tatactccca catatgccag attcagcaac ggatacggct tccccaactt
     2761 gcccacttcc atacgtgtcc tccttaccag aaatttatcc ttaagatccc gaatcgttta
     2821 aactcgactc tggctctatc gaatctccgt cgtttcgagc ttacgcgaac agccgtggcg
     2881 ctcatttgct cgtcgggcat cgaatctcgt cagctatcgt cagcttacct ttttggcagc
     2941 gatcgcggct cccgacatct tggaccatta gctccacagg tatcttcttc cctctagtgg
     3001 tcataacagc agcttcagct acctctcaat tcaaaaaacc cctcaagacc cgtttagagg
     3061 ccccaagggg ttatgctatc aatcgttgcg ttacacacac aaaaaaccaa cacacatcca
     3121 tcttcgatgg atagcgattt tattatctaa ctgctgatcg agtgtagcca gatctagtaa
     3181 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     3241 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     3301 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     3361 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     3421 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     3481 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatgct gatgcggttt
     3541 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
     3601 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     3661 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     3721 ataagcagag ctggtttagt gaaccgtcag atcagatctt tgtcgatcct accatccact
     3781 cgacacaccc gccagcggcc gccaccatgt ctgataatgg accacagaac caacgcaatg
     3841 caccccgaat cacttttggc ggtccgtccg acagtaccgg gagcaaccag aacggcgaaa
     3901 gatcgggagc tcgttcaaaa cagcggaggc ctcaagggtt acccaataac acggccagct
     3961 ggttcacagc gttgacccag catggcaagg aggacctcaa gttccctcgg ggtcaggggg
     4021 tcccaataaa tacaaacagt tcccctgatg accaaattgg atactataga cgcgctacta
     4081 ggaggatccg tggcggggat ggaaaaatga aggacctttc tcccagatgg tacttttatt
     4141 acctgggtac tggcccagag gccggtctac cgtatggagc aaacaaagat ggcattattt
     4201 gggtggccac ggaaggagca ctgaataccc caaaggatca catcggcacc cggaatccag
     4261 ccaacaatgc tgctatcgtt ctgcaactcc cccaggggac aacattgcct aaaggcttct
     4321 atgccgaggg cagtagaggt ggcagccagg cgtcctcacg cagctcttct cgatctcgga
     4381 acagctcaag aaattccact cccgggagtt cccgaggaac cagcccagca aggatggctg
     4441 gcaatggtgg agatgccgct ctggccctac ttctgctgga ccgcctcaat cagttagaat
     4501 caaagatgtc ggggaagggg cagcaacagc agggacagac cgtaactaaa aagagtgcag
     4561 cggaggcctc taaaaagcct cggcagaaac gtacggcaac aaaggcttac aacgtgaccc
     4621 aggcctttgg ccgtagagga cccgaacaga cacaagggaa ctttggtgac caagagttga
     4681 taaggcaggg aaccgactac aaacactggc ctcagatcgc ccaattcgca ccgagcgcga
     4741 gcgcattttt cggaatgtca cgaattggca tggaggtgac accttcggga acctggctga
     4801 catacactgg cgccataaag ctcgatgaca aagatcccaa cttcaaggat caagtcatct
     4861 tgctgaataa gcatattgac gcgtataaaa cttttccacc cacggaaccg aagaaggaca
     4921 agaaaaagaa agctgatgag acacaggcgc ttccacaaag gcagaaaaag cagcagacgg
     4981 ttactctact gcccgccgca gacctagatg acttctccaa acagcttcag cagtctatga
     5041 gtagcgccga ctcaacccaa gctgaaaacc tttactttca gtcgtacccc tatgacgtac
     5101 cagattatgc gtaaggatcc gcaggcctct gctagcttga ctgactgaga tacagcgtac
     5161 cttcagctca cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     5221 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     5281 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     5341 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tattggccca
     5401 tctctatcgg tatcgtagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     5461 tgtgcccctc gggccggatt gctatctacc ggcattggcg cagaaaaaaa tgcctgatgc
     5521 gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca
     5581 acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaagg tcgtcagcta
     5641 tcctgcaggc gatctctcga tttcgatcaa gacattcctt taatggtctt ttctggacac
     5701 cactaggggt cagaagtagt tcatcaaact ttcttccctc cctaatctca ttggttacct
     5761 tgggctatcg aaacttaatt aaccagtcaa gtcagctact tggcgagatc gacttgtctg
     5821 ggtttcgact acgctcagaa ttgcgtcagt caagttcgat ctggtccttg ctattgcacc
     5881 cgttctccga ttacgagttt catttaaatc atgtgagcaa aaggccagca aaaggccagg
     5941 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     6001 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     6061 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     6121 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     6181 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     6241 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     6301 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6361 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     6421 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6481 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     6541 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     6601 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6661 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     6721 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     6781 tcatccatag ttgcatttaa atttccgaac tctccaaggc cctcgtcgga aaatcttcaa
     6841 acctttcgtc cgatccatct tgcaggctac ctctcgaacg aactatcgca agtctcttgg
     6901 ccggccttgc gccttggcta ttgcttggca gcgcctatcg ccaggtatta ctccaatccc
     6961 gaatatccga gatcgggatc acccgagaga agttcaacct acatcctcaa tcccgatcta
     7021 tccgagatcc gaggaatatc gaaatcgggg cgcgcctggc ctccgcgccg ggttttggcg
     7081 cctcccgcgg gcgcccccct cgtcacggcg agcgctgcca cgtcagacga agggcgcagg
     7141 agcgtcctga tccttccgcc cggacgctca ggacagcggc ccgctgctca taagactcgg
     7201 ccttagaacc ccagtatcag cagaaggaca ttttaggacg ggacttgggt gactctaggg
     7261 cactggtttt ctttccagag agcggaacag gcgaggaaaa gtagtccctt ctcggcgatt
     7321 ctgcggaggg atctccgtgg ggcggtgaac gccgatgatt atataaggac gcgccgggtg
     7381 tggcacagct agttccgtcg cagccgggat ttgggtcgcg gttcttgttt gtggatcgct
     7441 gtgatcgtca cttggtgagt agcgggctgc tgggctggcc ggggctttcg tggccgccgg
     7501 gccgctcggt gggacggaag cgtgtggaga gaccgccaag ggctgtagtc tgggtccgcg
     7561 agcaaggttg ccctgaactg ggggttgggg ggagcgcagc aaaatggcgg ctgttcccga
     7621 gtcttgaatg gaagacgctt gtgaggcggg ctgtgaggtc gttgaaacaa ggtggggggc
     7681 atggtgggcg gcaagaaccc aaggtcttga gcccttcgct aatgcgggaa agctcttatt
     7741 cgggtgagat gggctgggca ccatctgggg accctgacgt gaagtttgtc actgactgga
     7801 gaactcggtt tgtcgtctgt tgcgggggcg gcagttatgg cggtg
//
