LOCUS       sequence-295604-        7889 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 7889)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7889)
  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-m4134518
FEATURES             Location/Qualifiers
     source          1..7889
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1262..1282
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1283..1360
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     polyA_signal    1429..1550
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1440..1459)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1494..1513
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1665..1711
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1741..1877
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1922..1993
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2219..2807)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2299..2318)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3307..4516
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4353..4372
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4520..5548
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5575..5623
                     /note="synthetic polyadenylation signal"
     CDS             join(5889..5957,5958..6749)
                     /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(6107..6126)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6762..6898
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7043..7114
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7119..7165)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7183..7231
                     /note="synthetic polyadenylation signal"
     enhancer        7312..7615
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7617..7819
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7770..7790
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgtct gataatggac cacagaacca acgcaatgca ccccgaatca cttttggcgg
       61 tccgtccgac agtaccggga gcaaccagaa cggcgaaaga tcgggagctc gttcaaaaca
      121 gcggaggcct caagggttac ccaataacac ggccagctgg ttcacagcgt tgacccagca
      181 tggcaaggag gacctcaagt tccctcgggg tcagggggtc ccaataaata caaacagttc
      241 ccctgatgac caaattggat actatagacg cgctactagg aggatccgtg gcggggatgg
      301 aaaaatgaag gacctttctc ccagatggta cttttattac ctgggtactg gcccagaggc
      361 cggtctaccg tatggagcaa acaaagatgg cattatttgg gtggccacgg aaggagcact
      421 gaatacccca aaggatcaca tcggcacccg gaatccagcc aacaatgctg ctatcgttct
      481 gcaactcccc caggggacaa cattgcctaa aggcttctat gccgagggca gtagaggtgg
      541 cagccaggcg tcctcacgca gctcttctcg atctcggaac agctcaagaa attccactcc
      601 cgggagttcc cgaggaacca gcccagcaag gatggctggc aatggtggag atgccgctct
      661 ggccctactt ctgctggacc gcctcaatca gttagaatca aagatgtcgg ggaaggggca
      721 gcaacagcag ggacagaccg taactaaaaa gagtgcagcg gaggcctcta aaaagcctcg
      781 gcagaaacgt acggcaacaa aggcttacaa cgtgacccag gcctttggcc gtagaggacc
      841 cgaacagaca caagggaact ttggtgacca agagttgata aggcagggaa ccgactacaa
      901 acactggcct cagatcgccc aattcgcacc gagcgcgagc gcatttttcg gaatgtcacg
      961 aattggcatg gaggtgacac cttcgggaac ctggctgaca tacactggcg ccataaagct
     1021 cgatgacaaa gatcccaact tcaaggatca agtcatcttg ctgaataagc atattgacgc
     1081 gtataaaact tttccaccca cggaaccgaa gaaggacaag aaaaagaaag ctgatgagac
     1141 acaggcgctt ccacaaaggc agaaaaagca gcagacggtt actctactgc ccgccgcaga
     1201 cctagatgac ttctccaaac agcttcagca gtctatgagt agcgccgact caacccaagc
     1261 tgaaaacctt tactttcagt cgaagaggcg atggaaaaag aattttatcg cggtgagtgc
     1321 agccaacaga ttcaaaaaga tatcctcgtc tggtgccctc taaggatccg caggcctctg
     1381 ctagcttgac tgactgagat acagcgtacc ttcagctcac agacatgata agatacattg
     1441 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     1501 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     1561 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
     1621 aaaacctcta caaatgtggt attggcccat ctctatcggt atcgtagcat aaccccttgg
     1681 ggcctctaaa cgggtcttga ggggtttttt gtgcccctcg ggccggattg ctatctaccg
     1741 gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc
     1801 cagattcagc aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac
     1861 cagaaattta tccttaaggt cgtcagctat cctgcaggcg atctctcgat ttcgatcaag
     1921 acattccttt aatggtcttt tctggacacc actaggggtc agaagtagtt catcaaactt
     1981 tcttccctcc ctaatctcat tggttacctt gggctatcga aacttaatta accagtcaag
     2041 tcagctactt ggcgagatcg acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc
     2101 aagttcgatc tggtccttgc tattgcaccc gttctccgat tacgagtttc atttaaatca
     2161 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2221 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2281 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2341 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2401 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     2461 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2521 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2581 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2641 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2701 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     2761 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2821 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     2881 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     2941 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3001 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcatttaaa tttccgaact
     3061 ctccaaggcc ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt gcaggctacc
     3121 tctcgaacga actatcgcaa gtctcttggc cggccttgcg ccttggctat tgcttggcag
     3181 cgcctatcgc caggtattac tccaatcccg aatatccgag atcgggatca cccgagagaa
     3241 gttcaaccta catcctcaat cccgatctat ccgagatccg aggaatatcg aaatcggggc
     3301 gcgcctggcc tccgcgccgg gttttggcgc ctcccgcggg cgcccccctc gtcacggcga
     3361 gcgctgccac gtcagacgaa gggcgcagga gcgtcctgat ccttccgccc ggacgctcag
     3421 gacagcggcc cgctgctcat aagactcggc cttagaaccc cagtatcagc agaaggacat
     3481 tttaggacgg gacttgggtg actctagggc actggttttc tttccagaga gcggaacagg
     3541 cgaggaaaag tagtcccttc tcggcgattc tgcggaggga tctccgtggg gcggtgaacg
     3601 ccgatgatta tataaggacg cgccgggtgt ggcacagcta gttccgtcgc agccgggatt
     3661 tgggtcgcgg ttcttgtttg tggatcgctg tgatcgtcac ttggtgagta gcgggctgct
     3721 gggctggccg gggctttcgt ggccgccggg ccgctcggtg ggacggaagc gtgtggagag
     3781 accgccaagg gctgtagtct gggtccgcga gcaaggttgc cctgaactgg gggttggggg
     3841 gagcgcagca aaatggcggc tgttcccgag tcttgaatgg aagacgcttg tgaggcgggc
     3901 tgtgaggtcg ttgaaacaag gtggggggca tggtgggcgg caagaaccca aggtcttgag
     3961 cccttcgcta atgcgggaaa gctcttattc gggtgagatg ggctgggcac catctgggga
     4021 ccctgacgtg aagtttgtca ctgactggag aactcggttt gtcgtctgtt gcgggggcgg
     4081 cagttatggc ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc gccctcgtcg
     4141 tgtcgtgacg tcacccgttc tgttggctta taatgcaggg tggggccacc tgccggtagg
     4201 tgtgcggtag gcttttctcc gtcgcaggac gcagggttcg ggcctagggt aggctctcct
     4261 gaatcgacag gcgccggacc tctggtgagg ggagggataa gtgaggcgtc agtttctttg
     4321 gtcggtttta tgtacctatc ttcttaagta gctgaagctc cggttttgaa ctatgcgctc
     4381 ggggttggcg agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt aatcatttgg
     4441 gtcaatatgt aattttcagt gttagacttg taaattgtcc gctaaattct ggccgttttt
     4501 ggcttttttg ttagacaaca tgggtaaaaa gcctgaactc accgcgacgt ctgtcgagaa
     4561 gtttctgatc gaaaagttcg acagcgtctc cgacctgatg cagctctcgg agggcgaaga
     4621 atctcgtgct ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg
     4681 cgccgatggt ttctacaaag atcgttatgt ttatcggcac tttgcatccg ccgcgctccc
     4741 gattccggaa gtgcttgaca ttggggagtt cagcgagagc ctgacctatt gcatctcccg
     4801 ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca
     4861 gccggtcgcg gaggcaatgg atgccatcgc tgccgccgat cttagccaga cgagcgggtt
     4921 cggcccattc ggaccgcaag gaatcggtca atacactaca tggcgtgatt tcatatgcgc
     4981 gattgctgat ccccatgtgt atcactggca aactgtgatg gacgacaccg tcagtgcgtc
     5041 cgtcgcgcag gctctcgatg agctgatgct ttgggccgag gactgccccg aagtccggca
     5101 cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg gacaatggcc gcataacagc
     5161 ggtcattgac tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt
     5221 cttctggagg ccgtggttgg cttgtatgga gcagcagacg cgctacttcg agcggaggca
     5281 tccggagctt gcaggatcgc cccggctccg ggcgtatatg ctccgcattg gtcttgacca
     5341 actctatcag agcttggttg acggcaattt cgatgatgca gcttgggcgc agggtcgatg
     5401 cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag
     5461 cgccgccgtc tggaccgatg gctgtgtaga agtactcgcc gatagtggaa accgacgccc
     5521 cagcactcgt ccgagggcaa aggaataagc tagtatgtaa gcctagtctt agataataaa
     5581 atcgctatcc atcgaagatg gatgtgtgtt ggttttttgt gtgtgtaacg ctaggcgcgc
     5641 ctggtgtacc gagaacgatc ctctcagtgc gagtctcgac gatccatatc gttgcttggc
     5701 agtcagccag tcggaatcca gcttgggacc caggaagtcc aatcgtcaga tattgtactc
     5761 aagcctggtc acggcagcgt accgatctgt ttaaacctag atattgatag tctgatcggt
     5821 caacgtataa tcgagtccta gcttttgcaa acatctatca agagacagga tcagcaggag
     5881 gctttcgcat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     5941 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     6001 tgggtgcgcg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     6061 ttcgccccga agaacgcttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     6121 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     6181 atgacttggt tgagtattca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     6241 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     6301 caacgattgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     6361 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     6421 ccacgatgcc tgtagcaatg gcaacaacct tgcgtaaact attaactggc gaactactta
     6481 ctctagcttc ccggcaacag ttgatagact ggatggaggc ggataaagtt gcaggaccac
     6541 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     6601 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     6661 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     6721 taggtgcctc actgattaag cattggtaac cgattctagg tgcattggcg cagaaaaaaa
     6781 tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac
     6841 ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaaga
     6901 tcgtttaaac tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta cgcgaacagc
     6961 cgtggcgctc atttgctcgt cgggcatcga atctcgtcag ctatcgtcag cttacctttt
     7021 tggcagcgat cgcggctccc gacatcttgg accattagct ccacaggtat cttcttccct
     7081 ctagtggtca taacagcagc ttcagctacc tctcaattca aaaaacccct caagacccgt
     7141 ttagaggccc caaggggtta tgctatcaat cgttgcgtta cacacacaaa aaaccaacac
     7201 acatccatct tcgatggata gcgattttat tatctaactg ctgatcgagt gtagccagat
     7261 ctagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     7321 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     7381 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     7441 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     7501 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     7561 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatgctgat
     7621 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
     7681 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     7741 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
     7801 ggtctatata agcagagctg gtttagtgaa ccgtcagatc agatctttgt cgatcctacc
     7861 atccactcga cacacccgcc agcggccgc
//
