LOCUS       sequence-307281-        7863 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 7863)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7863)
  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-m4133956
FEATURES             Location/Qualifiers
     source          1..7863
                     /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..1905
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    1974..2095
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1985..2004)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2039..2058
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2210..2256
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2286..2422
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2467..2538
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2764..3352)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2844..2863)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3852..5061
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4898..4917
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5065..6093
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    6120..6168
                     /note="synthetic polyadenylation signal"
     CDS             join(6434..6502,6503..7294)
                     /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(6652..6671)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7307..7443
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7595..7666
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7671..7717)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7735..7783
                     /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 aaagaaaccg cggcagctaa gttcgagagg cagcacatgg acagttaagg atccgcaggc
     1921 ctctgctagc ttgactgact gagatacagc gtaccttcag ctcacagaca tgataagata
     1981 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     2041 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     2101 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag
     2161 caagtaaaac ctctacaaat gtggtattgg cccatctcta tcggtatcgt agcataaccc
     2221 cttggggcct ctaaacgggt cttgaggggt tttttgtgcc cctcgggccg gattgctatc
     2281 taccggcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca
     2341 tatgccagat tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc
     2401 cttaccagaa atttatcctt aaggtcgtca gctatcctgc aggcgatctc tcgatttcga
     2461 tcaagacatt cctttaatgg tcttttctgg acaccactag gggtcagaag tagttcatca
     2521 aactttcttc cctccctaat ctcattggtt accttgggct atcgaaactt aattaaccag
     2581 tcaagtcagc tacttggcga gatcgacttg tctgggtttc gactacgctc agaattgcgt
     2641 cagtcaagtt cgatctggtc cttgctattg cacccgttct ccgattacga gtttcattta
     2701 aatcatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     2761 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     2821 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     2881 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2941 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3001 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3061 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3121 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3181 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     3241 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3301 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     3361 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3421 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     3481 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     3541 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcat ttaaatttcc
     3601 gaactctcca aggccctcgt cggaaaatct tcaaaccttt cgtccgatcc atcttgcagg
     3661 ctacctctcg aacgaactat cgcaagtctc ttggccggcc ttgcgccttg gctattgctt
     3721 ggcagcgcct atcgccaggt attactccaa tcccgaatat ccgagatcgg gatcacccga
     3781 gagaagttca acctacatcc tcaatcccga tctatccgag atccgaggaa tatcgaaatc
     3841 ggggcgcgcc tggcctccgc gccgggtttt ggcgcctccc gcgggcgccc ccctcgtcac
     3901 ggcgagcgct gccacgtcag acgaagggcg caggagcgtc ctgatccttc cgcccggacg
     3961 ctcaggacag cggcccgctg ctcataagac tcggccttag aaccccagta tcagcagaag
     4021 gacattttag gacgggactt gggtgactct agggcactgg ttttctttcc agagagcgga
     4081 acaggcgagg aaaagtagtc ccttctcggc gattctgcgg agggatctcc gtggggcggt
     4141 gaacgccgat gattatataa ggacgcgccg ggtgtggcac agctagttcc gtcgcagccg
     4201 ggatttgggt cgcggttctt gtttgtggat cgctgtgatc gtcacttggt gagtagcggg
     4261 ctgctgggct ggccggggct ttcgtggccg ccgggccgct cggtgggacg gaagcgtgtg
     4321 gagagaccgc caagggctgt agtctgggtc cgcgagcaag gttgccctga actgggggtt
     4381 ggggggagcg cagcaaaatg gcggctgttc ccgagtcttg aatggaagac gcttgtgagg
     4441 cgggctgtga ggtcgttgaa acaaggtggg gggcatggtg ggcggcaaga acccaaggtc
     4501 ttgagccctt cgctaatgcg ggaaagctct tattcgggtg agatgggctg ggcaccatct
     4561 ggggaccctg acgtgaagtt tgtcactgac tggagaactc ggtttgtcgt ctgttgcggg
     4621 ggcggcagtt atggcggtgc cgttgggcag tgcacccgta cctttgggag cgcgcgccct
     4681 cgtcgtgtcg tgacgtcacc cgttctgttg gcttataatg cagggtgggg ccacctgccg
     4741 gtaggtgtgc ggtaggcttt tctccgtcgc aggacgcagg gttcgggcct agggtaggct
     4801 ctcctgaatc gacaggcgcc ggacctctgg tgaggggagg gataagtgag gcgtcagttt
     4861 ctttggtcgg ttttatgtac ctatcttctt aagtagctga agctccggtt ttgaactatg
     4921 cgctcggggt tggcgagtgt gttttgtgaa gttttttagg caccttttga aatgtaatca
     4981 tttgggtcaa tatgtaattt tcagtgttag acttgtaaat tgtccgctaa attctggccg
     5041 tttttggctt ttttgttaga caacatgggt aaaaagcctg aactcaccgc gacgtctgtc
     5101 gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct ctcggagggc
     5161 gaagaatctc gtgctttcag cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat
     5221 agctgcgccg atggtttcta caaagatcgt tatgtttatc ggcactttgc atccgccgcg
     5281 ctcccgattc cggaagtgct tgacattggg gagttcagcg agagcctgac ctattgcatc
     5341 tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt
     5401 ctgcagccgg tcgcggaggc aatggatgcc atcgctgccg ccgatcttag ccagacgagc
     5461 gggttcggcc cattcggacc gcaaggaatc ggtcaataca ctacatggcg tgatttcata
     5521 tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg tgatggacga caccgtcagt
     5581 gcgtccgtcg cgcaggctct cgatgagctg atgctttggg ccgaggactg ccccgaagtc
     5641 cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa tggccgcata
     5701 acagcggtca ttgactggag cgaggcgatg ttcggggatt cccaatacga ggtcgccaac
     5761 atcttcttct ggaggccgtg gttggcttgt atggagcagc agacgcgcta cttcgagcgg
     5821 aggcatccgg agcttgcagg atcgccccgg ctccgggcgt atatgctccg cattggtctt
     5881 gaccaactct atcagagctt ggttgacggc aatttcgatg atgcagcttg ggcgcagggt
     5941 cgatgcgacg caatcgtccg atccggagcc gggactgtcg ggcgtacaca aatcgcccgc
     6001 agaagcgccg ccgtctggac cgatggctgt gtagaagtac tcgccgatag tggaaaccga
     6061 cgccccagca ctcgtccgag ggcaaaggaa taagctagta tgtaagccta gtcttagata
     6121 ataaaatcgc tatccatcga agatggatgt gtgttggttt tttgtgtgtg taacgctagg
     6181 cgcgcctggt gtaccgagaa cgatcctctc agtgcgagtc tcgacgatcc atatcgttgc
     6241 ttggcagtca gccagtcgga atccagcttg ggacccagga agtccaatcg tcagatattg
     6301 tactcaagcc tggtcacggc agcgtaccga tctgtttaaa cctagatatt gatagtctga
     6361 tcggtcaacg tataatcgag tcctagcttt tgcaaacatc tatcaagaga caggatcagc
     6421 aggaggcttt cgcatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     6481 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     6541 tcagttgggt gcgcgagtgg gttacatcga actggatctc aacagcggta agatccttga
     6601 gagttttcgc cccgaagaac gctttccaat gatgagcact tttaaagttc tgctatgtgg
     6661 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     6721 tcagaatgac ttggttgagt attcaccagt cacagaaaag catcttacgg atggcatgac
     6781 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     6841 tctgacaacg attggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     6901 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     6961 tgacaccacg atgcctgtag caatggcaac aaccttgcgt aaactattaa ctggcgaact
     7021 acttactcta gcttcccggc aacagttgat agactggatg gaggcggata aagttgcagg
     7081 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     7141 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     7201 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     7261 tgagataggt gcctcactga ttaagcattg gtaaccgatt ctaggtgcat tggcgcagaa
     7321 aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga ttcagcaacg
     7381 gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga aatttatcct
     7441 taagatcccg aatcgtttaa actcgactct ggctctatcg aatctccgtc gtttcgagct
     7501 tacgcgaaca gccgtggcgc tcatttgctc gtcgggcatc gaatctcgtc agctatcgtc
     7561 agcttacctt tttggcagcg atcgcggctc ccgacatctt ggaccattag ctccacaggt
     7621 atcttcttcc ctctagtggt cataacagca gcttcagcta cctctcaatt caaaaaaccc
     7681 ctcaagaccc gtttagaggc cccaaggggt tatgctatca atcgttgcgt tacacacaca
     7741 aaaaaccaac acacatccat cttcgatgga tagcgatttt attatctaac tgctgatcga
     7801 gtgtagccag atctagtaat caattacggg gtcattagtt catagcccat atatggagtt
     7861 ccg
//
