LOCUS       sequence-295570-        7829 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 7829)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7829)
  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..7829
                     /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..1300
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1369..1490
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1380..1399)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1434..1453
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1605..1651
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1681..1817
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1862..1933
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2159..2747)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2239..2258)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3247..4456
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4293..4312
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4460..5488
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5515..5563
                     /note="synthetic polyadenylation signal"
     CDS             join(5829..5897,5898..6689)
                     /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(6047..6066)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6702..6838
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6983..7054
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7059..7105)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7123..7171
                     /note="synthetic polyadenylation signal"
     enhancer        7252..7555
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7557..7759
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7710..7730
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgtct gataatggac cccaaaatca gcgaaatgca ccccgcatta cgtttggtgg
       61 accctcagat tcaactggca gtaaccagaa tggagaacgc agtggggcgc gatcaaaaca
      121 acgtcggccc caaggtttac ccaataatac tgcgtcttgg ttcaccgctc tcactcaaca
      181 tggcaaggaa gaccttaaat tccctcgagg acaaggcgtt ccaattaaca ccaatagcag
      241 tccagatgac caaattggct actaccgaag agctaccaga cgaattcgtg gtggtgacgg
      301 taaaatgaaa gatctcagtc caagatggta tttctactac ctaggaactg ggccagaagc
      361 tggacttccc tatggtgcta acaaagacgg catcatatgg gttgcaactg agggagcctt
      421 gaatacacca aaagatcaca ttggcacccg caatcctgct aacaatgctg caatcgtgct
      481 acaacttcct caaggaacaa cattgccaaa aggcttctac gcagaaggga gcagaggcgg
      541 cagtcaagcc tcttctcgtt cctcatcacg tagtcgcaac agttcaagaa attcaactcc
      601 aggcagcagt aggggaactt ctcctgctag aatggctggc aatggcggtg atgctgctct
      661 tgctttgctg ctgcttgaca gattgaacca gcttgagagc aaaatgtctg gtaaaggcca
      721 acaacaacaa ggccaaactg tcactaagaa atctgctgct gaggcttcta agaagcctcg
      781 gcaaaaacgt actgccacta aagcatacaa tgtaacacaa gctttcggca gacgtggtcc
      841 agaacaaacc caaggaaatt ttggggacca ggaactaatc agacaaggaa ctgattacaa
      901 acattggccg caaattgcac aatttgcccc cagcgcttca gcgttcttcg gaatgtcgcg
      961 cattggcatg gaagtcacac cttcgggaac gtggttgacc tacacaggtg ccatcaaatt
     1021 ggatgacaaa gatccaaatt tcaaagatca agtcattttg ctgaataagc atattgacgc
     1081 atacaaaaca ttcccaccaa cagagcctaa aaaggacaaa aagaagaagg ctgatgaaac
     1141 tcaagcctta ccgcagagac agaagaaaca gcaaactgtg actcttcttc ctgctgcaga
     1201 tttggatgat ttctccaaac aattgcaaca atccatgagc agtgctgact caactcaggc
     1261 cgaaaacctt tactttcagt cgcaccatca ccatcaccat taaggatccg caggcctctg
     1321 ctagcttgac tgactgagat acagcgtacc ttcagctcac agacatgata agatacattg
     1381 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     1441 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     1501 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
     1561 aaaacctcta caaatgtggt attggcccat ctctatcggt atcgtagcat aaccccttgg
     1621 ggcctctaaa cgggtcttga ggggtttttt gtgcccctcg ggccggattg ctatctaccg
     1681 gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc
     1741 cagattcagc aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac
     1801 cagaaattta tccttaaggt cgtcagctat cctgcaggcg atctctcgat ttcgatcaag
     1861 acattccttt aatggtcttt tctggacacc actaggggtc agaagtagtt catcaaactt
     1921 tcttccctcc ctaatctcat tggttacctt gggctatcga aacttaatta accagtcaag
     1981 tcagctactt ggcgagatcg acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc
     2041 aagttcgatc tggtccttgc tattgcaccc gttctccgat tacgagtttc atttaaatca
     2101 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2161 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2221 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2281 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2341 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     2401 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2461 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2521 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2581 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2641 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     2701 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2761 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     2821 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     2881 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     2941 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcatttaaa tttccgaact
     3001 ctccaaggcc ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt gcaggctacc
     3061 tctcgaacga actatcgcaa gtctcttggc cggccttgcg ccttggctat tgcttggcag
     3121 cgcctatcgc caggtattac tccaatcccg aatatccgag atcgggatca cccgagagaa
     3181 gttcaaccta catcctcaat cccgatctat ccgagatccg aggaatatcg aaatcggggc
     3241 gcgcctggcc tccgcgccgg gttttggcgc ctcccgcggg cgcccccctc gtcacggcga
     3301 gcgctgccac gtcagacgaa gggcgcagga gcgtcctgat ccttccgccc ggacgctcag
     3361 gacagcggcc cgctgctcat aagactcggc cttagaaccc cagtatcagc agaaggacat
     3421 tttaggacgg gacttgggtg actctagggc actggttttc tttccagaga gcggaacagg
     3481 cgaggaaaag tagtcccttc tcggcgattc tgcggaggga tctccgtggg gcggtgaacg
     3541 ccgatgatta tataaggacg cgccgggtgt ggcacagcta gttccgtcgc agccgggatt
     3601 tgggtcgcgg ttcttgtttg tggatcgctg tgatcgtcac ttggtgagta gcgggctgct
     3661 gggctggccg gggctttcgt ggccgccggg ccgctcggtg ggacggaagc gtgtggagag
     3721 accgccaagg gctgtagtct gggtccgcga gcaaggttgc cctgaactgg gggttggggg
     3781 gagcgcagca aaatggcggc tgttcccgag tcttgaatgg aagacgcttg tgaggcgggc
     3841 tgtgaggtcg ttgaaacaag gtggggggca tggtgggcgg caagaaccca aggtcttgag
     3901 cccttcgcta atgcgggaaa gctcttattc gggtgagatg ggctgggcac catctgggga
     3961 ccctgacgtg aagtttgtca ctgactggag aactcggttt gtcgtctgtt gcgggggcgg
     4021 cagttatggc ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc gccctcgtcg
     4081 tgtcgtgacg tcacccgttc tgttggctta taatgcaggg tggggccacc tgccggtagg
     4141 tgtgcggtag gcttttctcc gtcgcaggac gcagggttcg ggcctagggt aggctctcct
     4201 gaatcgacag gcgccggacc tctggtgagg ggagggataa gtgaggcgtc agtttctttg
     4261 gtcggtttta tgtacctatc ttcttaagta gctgaagctc cggttttgaa ctatgcgctc
     4321 ggggttggcg agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt aatcatttgg
     4381 gtcaatatgt aattttcagt gttagacttg taaattgtcc gctaaattct ggccgttttt
     4441 ggcttttttg ttagacaaca tgggtaaaaa gcctgaactc accgcgacgt ctgtcgagaa
     4501 gtttctgatc gaaaagttcg acagcgtctc cgacctgatg cagctctcgg agggcgaaga
     4561 atctcgtgct ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg
     4621 cgccgatggt ttctacaaag atcgttatgt ttatcggcac tttgcatccg ccgcgctccc
     4681 gattccggaa gtgcttgaca ttggggagtt cagcgagagc ctgacctatt gcatctcccg
     4741 ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca
     4801 gccggtcgcg gaggcaatgg atgccatcgc tgccgccgat cttagccaga cgagcgggtt
     4861 cggcccattc ggaccgcaag gaatcggtca atacactaca tggcgtgatt tcatatgcgc
     4921 gattgctgat ccccatgtgt atcactggca aactgtgatg gacgacaccg tcagtgcgtc
     4981 cgtcgcgcag gctctcgatg agctgatgct ttgggccgag gactgccccg aagtccggca
     5041 cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg gacaatggcc gcataacagc
     5101 ggtcattgac tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt
     5161 cttctggagg ccgtggttgg cttgtatgga gcagcagacg cgctacttcg agcggaggca
     5221 tccggagctt gcaggatcgc cccggctccg ggcgtatatg ctccgcattg gtcttgacca
     5281 actctatcag agcttggttg acggcaattt cgatgatgca gcttgggcgc agggtcgatg
     5341 cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag
     5401 cgccgccgtc tggaccgatg gctgtgtaga agtactcgcc gatagtggaa accgacgccc
     5461 cagcactcgt ccgagggcaa aggaataagc tagtatgtaa gcctagtctt agataataaa
     5521 atcgctatcc atcgaagatg gatgtgtgtt ggttttttgt gtgtgtaacg ctaggcgcgc
     5581 ctggtgtacc gagaacgatc ctctcagtgc gagtctcgac gatccatatc gttgcttggc
     5641 agtcagccag tcggaatcca gcttgggacc caggaagtcc aatcgtcaga tattgtactc
     5701 aagcctggtc acggcagcgt accgatctgt ttaaacctag atattgatag tctgatcggt
     5761 caacgtataa tcgagtccta gcttttgcaa acatctatca agagacagga tcagcaggag
     5821 gctttcgcat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     5881 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     5941 tgggtgcgcg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     6001 ttcgccccga agaacgcttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     6061 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     6121 atgacttggt tgagtattca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     6181 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     6241 caacgattgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     6301 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     6361 ccacgatgcc tgtagcaatg gcaacaacct tgcgtaaact attaactggc gaactactta
     6421 ctctagcttc ccggcaacag ttgatagact ggatggaggc ggataaagtt gcaggaccac
     6481 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     6541 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     6601 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     6661 taggtgcctc actgattaag cattggtaac cgattctagg tgcattggcg cagaaaaaaa
     6721 tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac
     6781 ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaaga
     6841 tcgtttaaac tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta cgcgaacagc
     6901 cgtggcgctc atttgctcgt cgggcatcga atctcgtcag ctatcgtcag cttacctttt
     6961 tggcagcgat cgcggctccc gacatcttgg accattagct ccacaggtat cttcttccct
     7021 ctagtggtca taacagcagc ttcagctacc tctcaattca aaaaacccct caagacccgt
     7081 ttagaggccc caaggggtta tgctatcaat cgttgcgtta cacacacaaa aaaccaacac
     7141 acatccatct tcgatggata gcgattttat tatctaactg ctgatcgagt gtagccagat
     7201 ctagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     7261 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     7321 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     7381 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     7441 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     7501 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatgctgat
     7561 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
     7621 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     7681 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
     7741 ggtctatata agcagagctg gtttagtgaa ccgtcagatc agatctttgt cgatcctacc
     7801 atccactcga cacacccgcc agcggccgc
//
