LOCUS       sequence-317273-        7836 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 7836)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7836)
  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-m4133784
FEATURES             Location/Qualifiers
     source          1..7836
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1840..1860
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1861..1878
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1947..2068
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1958..1977)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2012..2031
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2183..2229
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2259..2395
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2440..2511
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2737..3325)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2817..2836)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3825..5034
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4871..4890
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5038..6066
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    6093..6141
                     /note="synthetic polyadenylation signal"
     CDS             join(6407..6475,6476..7267)
                     /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(6625..6644)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7280..7416
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7568..7639
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7644..7690)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7708..7756
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
      541 gatcctacca tccactcgac acacccgcca gcggccgcca ccatgtctga taatggaccc
      601 caaaatcagc gaaatgcacc ccgcattacg tttggtggac cctcagattc aactggcagt
      661 aaccagaatg gagaacgcag tggggcgcga tcaaaacaac gtcggcccca aggtttaccc
      721 aataatactg cgtcttggtt caccgctctc actcaacatg gcaaggaaga ccttaaattc
      781 cctcgaggac aaggcgttcc aattaacacc aatagcagtc cagatgacca aattggctac
      841 taccgaagag ctaccagacg aattcgtggt ggtgacggta aaatgaaaga tctcagtcca
      901 agatggtatt tctactacct aggaactggg ccagaagctg gacttcccta tggtgctaac
      961 aaagacggca tcatatgggt tgcaactgag ggagccttga atacaccaaa agatcacatt
     1021 ggcacccgca atcctgctaa caatgctgca atcgtgctac aacttcctca aggaacaaca
     1081 ttgccaaaag gcttctacgc agaagggagc agaggcggca gtcaagcctc ttctcgttcc
     1141 tcatcacgta gtcgcaacag ttcaagaaat tcaactccag gcagcagtag gggaacttct
     1201 cctgctagaa tggctggcaa tggcggtgat gctgctcttg ctttgctgct gcttgacaga
     1261 ttgaaccagc ttgagagcaa aatgtctggt aaaggccaac aacaacaagg ccaaactgtc
     1321 actaagaaat ctgctgctga ggcttctaag aagcctcggc aaaaacgtac tgccactaaa
     1381 gcatacaatg taacacaagc tttcggcaga cgtggtccag aacaaaccca aggaaatttt
     1441 ggggaccagg aactaatcag acaaggaact gattacaaac attggccgca aattgcacaa
     1501 tttgccccca gcgcttcagc gttcttcgga atgtcgcgca ttggcatgga agtcacacct
     1561 tcgggaacgt ggttgaccta cacaggtgcc atcaaattgg atgacaaaga tccaaatttc
     1621 aaagatcaag tcattttgct gaataagcat attgacgcat acaaaacatt cccaccaaca
     1681 gagcctaaaa aggacaaaaa gaagaaggct gatgaaactc aagccttacc gcagagacag
     1741 aagaaacagc aaactgtgac tcttcttcct gctgcagatt tggatgattt ctccaaacaa
     1801 ttgcaacaat ccatgagcag tgctgactca actcaggccg aaaaccttta ctttcagtcg
     1861 caccatcacc atcaccatta aggatccgca ggcctctgct agcttgactg actgagatac
     1921 agcgtacctt cagctcacag acatgataag atacattgat gagtttggac aaaccacaac
     1981 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     2041 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     2101 ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat
     2161 tggcccatct ctatcggtat cgtagcataa ccccttgggg cctctaaacg ggtcttgagg
     2221 ggttttttgt gcccctcggg ccggattgct atctaccggc attggcgcag aaaaaaatgc
     2281 ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc
     2341 ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaaggtcg
     2401 tcagctatcc tgcaggcgat ctctcgattt cgatcaagac attcctttaa tggtcttttc
     2461 tggacaccac taggggtcag aagtagttca tcaaactttc ttccctccct aatctcattg
     2521 gttaccttgg gctatcgaaa cttaattaac cagtcaagtc agctacttgg cgagatcgac
     2581 ttgtctgggt ttcgactacg ctcagaattg cgtcagtcaa gttcgatctg gtccttgcta
     2641 ttgcacccgt tctccgatta cgagtttcat ttaaatcatg tgagcaaaag gccagcaaaa
     2701 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2761 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2821 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2881 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2941 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3001 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3061 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3121 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     3181 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3241 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3301 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3361 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3421 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3481 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     3541 atttcgttca tccatagttg catttaaatt tccgaactct ccaaggccct cgtcggaaaa
     3601 tcttcaaacc tttcgtccga tccatcttgc aggctacctc tcgaacgaac tatcgcaagt
     3661 ctcttggccg gccttgcgcc ttggctattg cttggcagcg cctatcgcca ggtattactc
     3721 caatcccgaa tatccgagat cgggatcacc cgagagaagt tcaacctaca tcctcaatcc
     3781 cgatctatcc gagatccgag gaatatcgaa atcggggcgc gcctggcctc cgcgccgggt
     3841 tttggcgcct cccgcgggcg cccccctcgt cacggcgagc gctgccacgt cagacgaagg
     3901 gcgcaggagc gtcctgatcc ttccgcccgg acgctcagga cagcggcccg ctgctcataa
     3961 gactcggcct tagaacccca gtatcagcag aaggacattt taggacggga cttgggtgac
     4021 tctagggcac tggttttctt tccagagagc ggaacaggcg aggaaaagta gtcccttctc
     4081 ggcgattctg cggagggatc tccgtggggc ggtgaacgcc gatgattata taaggacgcg
     4141 ccgggtgtgg cacagctagt tccgtcgcag ccgggatttg ggtcgcggtt cttgtttgtg
     4201 gatcgctgtg atcgtcactt ggtgagtagc gggctgctgg gctggccggg gctttcgtgg
     4261 ccgccgggcc gctcggtggg acggaagcgt gtggagagac cgccaagggc tgtagtctgg
     4321 gtccgcgagc aaggttgccc tgaactgggg gttgggggga gcgcagcaaa atggcggctg
     4381 ttcccgagtc ttgaatggaa gacgcttgtg aggcgggctg tgaggtcgtt gaaacaaggt
     4441 ggggggcatg gtgggcggca agaacccaag gtcttgagcc cttcgctaat gcgggaaagc
     4501 tcttattcgg gtgagatggg ctgggcacca tctggggacc ctgacgtgaa gtttgtcact
     4561 gactggagaa ctcggtttgt cgtctgttgc gggggcggca gttatggcgg tgccgttggg
     4621 cagtgcaccc gtacctttgg gagcgcgcgc cctcgtcgtg tcgtgacgtc acccgttctg
     4681 ttggcttata atgcagggtg gggccacctg ccggtaggtg tgcggtaggc ttttctccgt
     4741 cgcaggacgc agggttcggg cctagggtag gctctcctga atcgacaggc gccggacctc
     4801 tggtgagggg agggataagt gaggcgtcag tttctttggt cggttttatg tacctatctt
     4861 cttaagtagc tgaagctccg gttttgaact atgcgctcgg ggttggcgag tgtgttttgt
     4921 gaagtttttt aggcaccttt tgaaatgtaa tcatttgggt caatatgtaa ttttcagtgt
     4981 tagacttgta aattgtccgc taaattctgg ccgtttttgg cttttttgtt agacaacatg
     5041 ggtaaaaagc ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac
     5101 agcgtctccg acctgatgca gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat
     5161 gtaggagggc gtggatatgt cctgcgggta aatagctgcg ccgatggttt ctacaaagat
     5221 cgttatgttt atcggcactt tgcatccgcc gcgctcccga ttccggaagt gcttgacatt
     5281 ggggagttca gcgagagcct gacctattgc atctcccgcc gtgcacaggg tgtcacgttg
     5341 caagacctgc ctgaaaccga actgcccgct gttctgcagc cggtcgcgga ggcaatggat
     5401 gccatcgctg ccgccgatct tagccagacg agcgggttcg gcccattcgg accgcaagga
     5461 atcggtcaat acactacatg gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat
     5521 cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag
     5581 ctgatgcttt gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc
     5641 tccaacaatg tcctgacgga caatggccgc ataacagcgg tcattgactg gagcgaggcg
     5701 atgttcgggg attcccaata cgaggtcgcc aacatcttct tctggaggcc gtggttggct
     5761 tgtatggagc agcagacgcg ctacttcgag cggaggcatc cggagcttgc aggatcgccc
     5821 cggctccggg cgtatatgct ccgcattggt cttgaccaac tctatcagag cttggttgac
     5881 ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga
     5941 gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg ccgccgtctg gaccgatggc
     6001 tgtgtagaag tactcgccga tagtggaaac cgacgcccca gcactcgtcc gagggcaaag
     6061 gaataagcta gtatgtaagc ctagtcttag ataataaaat cgctatccat cgaagatgga
     6121 tgtgtgttgg ttttttgtgt gtgtaacgct aggcgcgcct ggtgtaccga gaacgatcct
     6181 ctcagtgcga gtctcgacga tccatatcgt tgcttggcag tcagccagtc ggaatccagc
     6241 ttgggaccca ggaagtccaa tcgtcagata ttgtactcaa gcctggtcac ggcagcgtac
     6301 cgatctgttt aaacctagat attgatagtc tgatcggtca acgtataatc gagtcctagc
     6361 ttttgcaaac atctatcaag agacaggatc agcaggaggc tttcgcatga gtattcaaca
     6421 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     6481 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcgcgag tgggttacat
     6541 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgctttcc
     6601 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     6661 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtattcacc
     6721 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     6781 aaccatgagt gataacactg cggccaactt acttctgaca acgattggag gaccgaagga
     6841 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     6901 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     6961 aacaaccttg cgtaaactat taactggcga actacttact ctagcttccc ggcaacagtt
     7021 gatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     7081 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     7141 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     7201 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     7261 ttggtaaccg attctaggtg cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt
     7321 cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac ttgcccactt
     7381 ccatacgtgt cctccttacc agaaatttat ccttaagatc ccgaatcgtt taaactcgac
     7441 tctggctcta tcgaatctcc gtcgtttcga gcttacgcga acagccgtgg cgctcatttg
     7501 ctcgtcgggc atcgaatctc gtcagctatc gtcagcttac ctttttggca gcgatcgcgg
     7561 ctcccgacat cttggaccat tagctccaca ggtatcttct tccctctagt ggtcataaca
     7621 gcagcttcag ctacctctca attcaaaaaa cccctcaaga cccgtttaga ggccccaagg
     7681 ggttatgcta tcaatcgttg cgttacacac acaaaaaacc aacacacatc catcttcgat
     7741 ggatagcgat tttattatct aactgctgat cgagtgtagc cagatctagt aatcaattac
     7801 ggggtcatta gttcatagcc catatatgga gttccg
//
