LOCUS       sequence-306186-        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-m4134525
FEATURES             Location/Qualifiers
     source          1..7836
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    103..174
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(179..225)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    243..291
                     /note="synthetic polyadenylation signal"
     enhancer        372..675
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        677..879
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     830..850
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2211..2231
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2232..2249
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2318..2439
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2329..2348)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2383..2402
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2554..2600
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2630..2766
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2811..2882
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3108..3696)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3188..3207)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4196..5405
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5242..5261
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5409..6437
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    6464..6512
                     /note="synthetic polyadenylation signal"
     CDS             join(6778..6846,6847..7638)
                     /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(6996..7015)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7651..7787
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 ttcgagctta cgcgaacagc cgtggcgctc atttgctcgt cgggcatcga atctcgtcag
       61 ctatcgtcag cttacctttt tggcagcgat cgcggctccc gacatcttgg accattagct
      121 ccacaggtat cttcttccct ctagtggtca taacagcagc ttcagctacc tctcaattca
      181 aaaaacccct caagacccgt ttagaggccc caaggggtta tgctatcaat cgttgcgtta
      241 cacacacaaa aaaccaacac acatccatct tcgatggata gcgattttat tatctaactg
      301 ctgatcgagt gtagccagat ctagtaatca attacggggt cattagttca tagcccatat
      361 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
      421 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      481 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      541 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      601 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      661 atcgctatta ccatgctgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      721 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      781 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
      841 ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc
      901 agatctttgt cgatcctacc atccactcga cacacccgcc agcggccgcc accatgtctg
      961 ataatggacc acagaaccaa cgcaatgcac cccgaatcac ttttggcggt ccgtccgaca
     1021 gtaccgggag caaccagaac ggcgaaagat cgggagctcg ttcaaaacag cggaggcctc
     1081 aagggttacc caataacacg gccagctggt tcacagcgtt gacccagcat ggcaaggagg
     1141 acctcaagtt ccctcggggt cagggggtcc caataaatac aaacagttcc cctgatgacc
     1201 aaattggata ctatagacgc gctactagga ggatccgtgg cggggatgga aaaatgaagg
     1261 acctttctcc cagatggtac ttttattacc tgggtactgg cccagaggcc ggtctaccgt
     1321 atggagcaaa caaagatggc attatttggg tggccacgga aggagcactg aataccccaa
     1381 aggatcacat cggcacccgg aatccagcca acaatgctgc tatcgttctg caactccccc
     1441 aggggacaac attgcctaaa ggcttctatg ccgagggcag tagaggtggc agccaggcgt
     1501 cctcacgcag ctcttctcga tctcggaaca gctcaagaaa ttccactccc gggagttccc
     1561 gaggaaccag cccagcaagg atggctggca atggtggaga tgccgctctg gccctacttc
     1621 tgctggaccg cctcaatcag ttagaatcaa agatgtcggg gaaggggcag caacagcagg
     1681 gacagaccgt aactaaaaag agtgcagcgg aggcctctaa aaagcctcgg cagaaacgta
     1741 cggcaacaaa ggcttacaac gtgacccagg cctttggccg tagaggaccc gaacagacac
     1801 aagggaactt tggtgaccaa gagttgataa ggcagggaac cgactacaaa cactggcctc
     1861 agatcgccca attcgcaccg agcgcgagcg catttttcgg aatgtcacga attggcatgg
     1921 aggtgacacc ttcgggaacc tggctgacat acactggcgc cataaagctc gatgacaaag
     1981 atcccaactt caaggatcaa gtcatcttgc tgaataagca tattgacgcg tataaaactt
     2041 ttccacccac ggaaccgaag aaggacaaga aaaagaaagc tgatgagaca caggcgcttc
     2101 cacaaaggca gaaaaagcag cagacggtta ctctactgcc cgccgcagac ctagatgact
     2161 tctccaaaca gcttcagcag tctatgagta gcgccgactc aacccaagct gaaaaccttt
     2221 actttcagtc gcaccatcac catcaccatt aaggatccgc aggcctctgc tagcttgact
     2281 gactgagata cagcgtacct tcagctcaca gacatgataa gatacattga tgagtttgga
     2341 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     2401 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     2461 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac
     2521 aaatgtggta ttggcccatc tctatcggta tcgtagcata accccttggg gcctctaaac
     2581 gggtcttgag gggttttttg tgcccctcgg gccggattgc tatctaccgg cattggcgca
     2641 gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc agattcagca
     2701 acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc agaaatttat
     2761 ccttaaggtc gtcagctatc ctgcaggcga tctctcgatt tcgatcaaga cattccttta
     2821 atggtctttt ctggacacca ctaggggtca gaagtagttc atcaaacttt cttccctccc
     2881 taatctcatt ggttaccttg ggctatcgaa acttaattaa ccagtcaagt cagctacttg
     2941 gcgagatcga cttgtctggg tttcgactac gctcagaatt gcgtcagtca agttcgatct
     3001 ggtccttgct attgcacccg ttctccgatt acgagtttca tttaaatcat gtgagcaaaa
     3061 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     3121 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3181 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     3241 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     3301 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     3361 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3421 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     3481 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     3541 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     3601 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     3661 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     3721 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     3781 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     3841 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     3901 gcgatctgtc tatttcgttc atccatagtt gcatttaaat ttccgaactc tccaaggccc
     3961 tcgtcggaaa atcttcaaac ctttcgtccg atccatcttg caggctacct ctcgaacgaa
     4021 ctatcgcaag tctcttggcc ggccttgcgc cttggctatt gcttggcagc gcctatcgcc
     4081 aggtattact ccaatcccga atatccgaga tcgggatcac ccgagagaag ttcaacctac
     4141 atcctcaatc ccgatctatc cgagatccga ggaatatcga aatcggggcg cgcctggcct
     4201 ccgcgccggg ttttggcgcc tcccgcgggc gcccccctcg tcacggcgag cgctgccacg
     4261 tcagacgaag ggcgcaggag cgtcctgatc cttccgcccg gacgctcagg acagcggccc
     4321 gctgctcata agactcggcc ttagaacccc agtatcagca gaaggacatt ttaggacggg
     4381 acttgggtga ctctagggca ctggttttct ttccagagag cggaacaggc gaggaaaagt
     4441 agtcccttct cggcgattct gcggagggat ctccgtgggg cggtgaacgc cgatgattat
     4501 ataaggacgc gccgggtgtg gcacagctag ttccgtcgca gccgggattt gggtcgcggt
     4561 tcttgtttgt ggatcgctgt gatcgtcact tggtgagtag cgggctgctg ggctggccgg
     4621 ggctttcgtg gccgccgggc cgctcggtgg gacggaagcg tgtggagaga ccgccaaggg
     4681 ctgtagtctg ggtccgcgag caaggttgcc ctgaactggg ggttgggggg agcgcagcaa
     4741 aatggcggct gttcccgagt cttgaatgga agacgcttgt gaggcgggct gtgaggtcgt
     4801 tgaaacaagg tggggggcat ggtgggcggc aagaacccaa ggtcttgagc ccttcgctaa
     4861 tgcgggaaag ctcttattcg ggtgagatgg gctgggcacc atctggggac cctgacgtga
     4921 agtttgtcac tgactggaga actcggtttg tcgtctgttg cgggggcggc agttatggcg
     4981 gtgccgttgg gcagtgcacc cgtacctttg ggagcgcgcg ccctcgtcgt gtcgtgacgt
     5041 cacccgttct gttggcttat aatgcagggt ggggccacct gccggtaggt gtgcggtagg
     5101 cttttctccg tcgcaggacg cagggttcgg gcctagggta ggctctcctg aatcgacagg
     5161 cgccggacct ctggtgaggg gagggataag tgaggcgtca gtttctttgg tcggttttat
     5221 gtacctatct tcttaagtag ctgaagctcc ggttttgaac tatgcgctcg gggttggcga
     5281 gtgtgttttg tgaagttttt taggcacctt ttgaaatgta atcatttggg tcaatatgta
     5341 attttcagtg ttagacttgt aaattgtccg ctaaattctg gccgtttttg gcttttttgt
     5401 tagacaacat gggtaaaaag cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg
     5461 aaaagttcga cagcgtctcc gacctgatgc agctctcgga gggcgaagaa tctcgtgctt
     5521 tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc gccgatggtt
     5581 tctacaaaga tcgttatgtt tatcggcact ttgcatccgc cgcgctcccg attccggaag
     5641 tgcttgacat tggggagttc agcgagagcc tgacctattg catctcccgc cgtgcacagg
     5701 gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg
     5761 aggcaatgga tgccatcgct gccgccgatc ttagccagac gagcgggttc ggcccattcg
     5821 gaccgcaagg aatcggtcaa tacactacat ggcgtgattt catatgcgcg attgctgatc
     5881 cccatgtgta tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg
     5941 ctctcgatga gctgatgctt tgggccgagg actgccccga agtccggcac ctcgtgcacg
     6001 cggatttcgg ctccaacaat gtcctgacgg acaatggccg cataacagcg gtcattgact
     6061 ggagcgaggc gatgttcggg gattcccaat acgaggtcgc caacatcttc ttctggaggc
     6121 cgtggttggc ttgtatggag cagcagacgc gctacttcga gcggaggcat ccggagcttg
     6181 caggatcgcc ccggctccgg gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga
     6241 gcttggttga cggcaatttc gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg
     6301 tccgatccgg agccgggact gtcgggcgta cacaaatcgc ccgcagaagc gccgccgtct
     6361 ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc agcactcgtc
     6421 cgagggcaaa ggaataagct agtatgtaag cctagtctta gataataaaa tcgctatcca
     6481 tcgaagatgg atgtgtgttg gttttttgtg tgtgtaacgc taggcgcgcc tggtgtaccg
     6541 agaacgatcc tctcagtgcg agtctcgacg atccatatcg ttgcttggca gtcagccagt
     6601 cggaatccag cttgggaccc aggaagtcca atcgtcagat attgtactca agcctggtca
     6661 cggcagcgta ccgatctgtt taaacctaga tattgatagt ctgatcggtc aacgtataat
     6721 cgagtcctag cttttgcaaa catctatcaa gagacaggat cagcaggagg ctttcgcatg
     6781 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     6841 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcgcga
     6901 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     6961 gaacgctttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     7021 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     7081 gagtattcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     7141 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgattgga
     7201 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     7261 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     7321 gtagcaatgg caacaacctt gcgtaaacta ttaactggcg aactacttac tctagcttcc
     7381 cggcaacagt tgatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     7441 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     7501 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     7561 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     7621 ctgattaagc attggtaacc gattctaggt gcattggcgc agaaaaaaat gcctgatgcg
     7681 acgctgcgcg tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa
     7741 cttgcccact tccatacgtg tcctccttac cagaaattta tccttaagat cccgaatcgt
     7801 ttaaactcga ctctggctct atcgaatctc cgtcgt
//
