LOCUS       sequence-306182-        7860 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 7860)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7860)
  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-m4134545
FEATURES             Location/Qualifiers
     source          1..7860
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    67..138
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(143..189)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    207..255
                     /note="synthetic polyadenylation signal"
     enhancer        336..639
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        641..843
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     794..814
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2175..2195
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2196..2237
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    2306..2427
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2317..2336)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2371..2390
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2542..2588
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2618..2754
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2799..2870
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3096..3684)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3176..3195)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4184..5393
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5230..5249
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5397..6425
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    6452..6500
                     /note="synthetic polyadenylation signal"
     CDS             join(6766..6834,6835..7626)
                     /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(6984..7003)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7639..7775
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 tcgtcgggca tcgaatctcg tcagctatcg tcagcttacc tttttggcag cgatcgcggc
       61 tcccgacatc ttggaccatt agctccacag gtatcttctt ccctctagtg gtcataacag
      121 cagcttcagc tacctctcaa ttcaaaaaac ccctcaagac ccgtttagag gccccaaggg
      181 gttatgctat caatcgttgc gttacacaca caaaaaacca acacacatcc atcttcgatg
      241 gatagcgatt ttattatcta actgctgatc gagtgtagcc agatctagta atcaattacg
      301 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
      361 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
      421 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
      481 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
      541 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
      601 tggcagtaca tctacgtatt agtcatcgct attaccatgc tgatgcggtt ttggcagtac
      661 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
      721 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
      781 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
      841 gctggtttag tgaaccgtca gatcagatct ttgtcgatcc taccatccac tcgacacacc
      901 cgccagcggc cgccaccatg tctgataatg gaccacagaa ccaacgcaat gcaccccgaa
      961 tcacttttgg cggtccgtcc gacagtaccg ggagcaacca gaacggcgaa agatcgggag
     1021 ctcgttcaaa acagcggagg cctcaagggt tacccaataa cacggccagc tggttcacag
     1081 cgttgaccca gcatggcaag gaggacctca agttccctcg gggtcagggg gtcccaataa
     1141 atacaaacag ttcccctgat gaccaaattg gatactatag acgcgctact aggaggatcc
     1201 gtggcgggga tggaaaaatg aaggaccttt ctcccagatg gtacttttat tacctgggta
     1261 ctggcccaga ggccggtcta ccgtatggag caaacaaaga tggcattatt tgggtggcca
     1321 cggaaggagc actgaatacc ccaaaggatc acatcggcac ccggaatcca gccaacaatg
     1381 ctgctatcgt tctgcaactc ccccagggga caacattgcc taaaggcttc tatgccgagg
     1441 gcagtagagg tggcagccag gcgtcctcac gcagctcttc tcgatctcgg aacagctcaa
     1501 gaaattccac tcccgggagt tcccgaggaa ccagcccagc aaggatggct ggcaatggtg
     1561 gagatgccgc tctggcccta cttctgctgg accgcctcaa tcagttagaa tcaaagatgt
     1621 cggggaaggg gcagcaacag cagggacaga ccgtaactaa aaagagtgca gcggaggcct
     1681 ctaaaaagcc tcggcagaaa cgtacggcaa caaaggctta caacgtgacc caggcctttg
     1741 gccgtagagg acccgaacag acacaaggga actttggtga ccaagagttg ataaggcagg
     1801 gaaccgacta caaacactgg cctcagatcg cccaattcgc accgagcgcg agcgcatttt
     1861 tcggaatgtc acgaattggc atggaggtga caccttcggg aacctggctg acatacactg
     1921 gcgccataaa gctcgatgac aaagatccca acttcaagga tcaagtcatc ttgctgaata
     1981 agcatattga cgcgtataaa acttttccac ccacggaacc gaagaaggac aagaaaaaga
     2041 aagctgatga gacacaggcg cttccacaaa ggcagaaaaa gcagcagacg gttactctac
     2101 tgcccgccgc agacctagat gacttctcca aacagcttca gcagtctatg agtagcgccg
     2161 actcaaccca agctgaaaac ctttactttc agtcggggaa acctataccc aatccacttc
     2221 tcggcttaga ttctacctaa ggatccgcag gcctctgcta gcttgactga ctgagataca
     2281 gcgtaccttc agctcacaga catgataaga tacattgatg agtttggaca aaccacaact
     2341 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     2401 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     2461 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatt
     2521 ggcccatctc tatcggtatc gtagcataac cccttggggc ctctaaacgg gtcttgaggg
     2581 gttttttgtg cccctcgggc cggattgcta tctaccggca ttggcgcaga aaaaaatgcc
     2641 tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac ggatacggct
     2701 tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc ttaaggtcgt
     2761 cagctatcct gcaggcgatc tctcgatttc gatcaagaca ttcctttaat ggtcttttct
     2821 ggacaccact aggggtcaga agtagttcat caaactttct tccctcccta atctcattgg
     2881 ttaccttggg ctatcgaaac ttaattaacc agtcaagtca gctacttggc gagatcgact
     2941 tgtctgggtt tcgactacgc tcagaattgc gtcagtcaag ttcgatctgg tccttgctat
     3001 tgcacccgtt ctccgattac gagtttcatt taaatcatgt gagcaaaagg ccagcaaaag
     3061 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3121 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3181 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3241 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3301 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3361 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3421 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3481 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     3541 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3601 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3661 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3721 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     3781 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     3841 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     3901 tttcgttcat ccatagttgc atttaaattt ccgaactctc caaggccctc gtcggaaaat
     3961 cttcaaacct ttcgtccgat ccatcttgca ggctacctct cgaacgaact atcgcaagtc
     4021 tcttggccgg ccttgcgcct tggctattgc ttggcagcgc ctatcgccag gtattactcc
     4081 aatcccgaat atccgagatc gggatcaccc gagagaagtt caacctacat cctcaatccc
     4141 gatctatccg agatccgagg aatatcgaaa tcggggcgcg cctggcctcc gcgccgggtt
     4201 ttggcgcctc ccgcgggcgc ccccctcgtc acggcgagcg ctgccacgtc agacgaaggg
     4261 cgcaggagcg tcctgatcct tccgcccgga cgctcaggac agcggcccgc tgctcataag
     4321 actcggcctt agaaccccag tatcagcaga aggacatttt aggacgggac ttgggtgact
     4381 ctagggcact ggttttcttt ccagagagcg gaacaggcga ggaaaagtag tcccttctcg
     4441 gcgattctgc ggagggatct ccgtggggcg gtgaacgccg atgattatat aaggacgcgc
     4501 cgggtgtggc acagctagtt ccgtcgcagc cgggatttgg gtcgcggttc ttgtttgtgg
     4561 atcgctgtga tcgtcacttg gtgagtagcg ggctgctggg ctggccgggg ctttcgtggc
     4621 cgccgggccg ctcggtggga cggaagcgtg tggagagacc gccaagggct gtagtctggg
     4681 tccgcgagca aggttgccct gaactggggg ttggggggag cgcagcaaaa tggcggctgt
     4741 tcccgagtct tgaatggaag acgcttgtga ggcgggctgt gaggtcgttg aaacaaggtg
     4801 gggggcatgg tgggcggcaa gaacccaagg tcttgagccc ttcgctaatg cgggaaagct
     4861 cttattcggg tgagatgggc tgggcaccat ctggggaccc tgacgtgaag tttgtcactg
     4921 actggagaac tcggtttgtc gtctgttgcg ggggcggcag ttatggcggt gccgttgggc
     4981 agtgcacccg tacctttggg agcgcgcgcc ctcgtcgtgt cgtgacgtca cccgttctgt
     5041 tggcttataa tgcagggtgg ggccacctgc cggtaggtgt gcggtaggct tttctccgtc
     5101 gcaggacgca gggttcgggc ctagggtagg ctctcctgaa tcgacaggcg ccggacctct
     5161 ggtgagggga gggataagtg aggcgtcagt ttctttggtc ggttttatgt acctatcttc
     5221 ttaagtagct gaagctccgg ttttgaacta tgcgctcggg gttggcgagt gtgttttgtg
     5281 aagtttttta ggcacctttt gaaatgtaat catttgggtc aatatgtaat tttcagtgtt
     5341 agacttgtaa attgtccgct aaattctggc cgtttttggc ttttttgtta gacaacatgg
     5401 gtaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca
     5461 gcgtctccga cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg
     5521 taggagggcg tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc
     5581 gttatgttta tcggcacttt gcatccgccg cgctcccgat tccggaagtg cttgacattg
     5641 gggagttcag cgagagcctg acctattgca tctcccgccg tgcacagggt gtcacgttgc
     5701 aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag gcaatggatg
     5761 ccatcgctgc cgccgatctt agccagacga gcgggttcgg cccattcgga ccgcaaggaa
     5821 tcggtcaata cactacatgg cgtgatttca tatgcgcgat tgctgatccc catgtgtatc
     5881 actggcaaac tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc
     5941 tgatgctttg ggccgaggac tgccccgaag tccggcacct cgtgcacgcg gatttcggct
     6001 ccaacaatgt cctgacggac aatggccgca taacagcggt cattgactgg agcgaggcga
     6061 tgttcgggga ttcccaatac gaggtcgcca acatcttctt ctggaggccg tggttggctt
     6121 gtatggagca gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgcccc
     6181 ggctccgggc gtatatgctc cgcattggtc ttgaccaact ctatcagagc ttggttgacg
     6241 gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag
     6301 ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc cgccgtctgg accgatggct
     6361 gtgtagaagt actcgccgat agtggaaacc gacgccccag cactcgtccg agggcaaagg
     6421 aataagctag tatgtaagcc tagtcttaga taataaaatc gctatccatc gaagatggat
     6481 gtgtgttggt tttttgtgtg tgtaacgcta ggcgcgcctg gtgtaccgag aacgatcctc
     6541 tcagtgcgag tctcgacgat ccatatcgtt gcttggcagt cagccagtcg gaatccagct
     6601 tgggacccag gaagtccaat cgtcagatat tgtactcaag cctggtcacg gcagcgtacc
     6661 gatctgttta aacctagata ttgatagtct gatcggtcaa cgtataatcg agtcctagct
     6721 tttgcaaaca tctatcaaga gacaggatca gcaggaggct ttcgcatgag tattcaacat
     6781 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     6841 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcgcgagt gggttacatc
     6901 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgctttcca
     6961 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     7021 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtattcacca
     7081 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     7141 accatgagtg ataacactgc ggccaactta cttctgacaa cgattggagg accgaaggag
     7201 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     7261 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     7321 acaaccttgc gtaaactatt aactggcgaa ctacttactc tagcttcccg gcaacagttg
     7381 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     7441 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     7501 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     7561 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     7621 tggtaaccga ttctaggtgc attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc
     7681 ttatactccc acatatgcca gattcagcaa cggatacggc ttccccaact tgcccacttc
     7741 catacgtgtc ctccttacca gaaatttatc cttaagatcc cgaatcgttt aaactcgact
     7801 ctggctctat cgaatctccg tcgtttcgag cttacgcgaa cagccgtggc gctcatttgc
//
