LOCUS       sequence-295506-        5484 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 5484)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5484)
  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-m4133747
FEATURES             Location/Qualifiers
     source          1..5484
                     /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    1353..1474
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1364..1383)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1418..1437
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1589..1635
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1665..1801
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1846..1917
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2143..2731)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2223..2242)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3477..3545,3546..4337)
                     /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(3695..3714)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4350..4486
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4638..4709
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4714..4760)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4778..4826
                     /note="synthetic polyadenylation signal"
     enhancer        4907..5210
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5212..5414
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5365..5385
                     /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 taagctagct tgactgactg
     1321 agatacagcg taccttcagc tcacagacat gataagatac attgatgagt ttggacaaac
     1381 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     1441 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     1501 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     1561 tggtattggc ccatctctat cggtatcgta gcataacccc ttggggcctc taaacgggtc
     1621 ttgaggggtt ttttgtgccc ctcgggccgg attgctatct accggcattg gcgcagaaaa
     1681 aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga
     1741 tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta
     1801 aggtcgtcag ctatcctgca ggcgatctct cgatttcgat caagacattc ctttaatggt
     1861 cttttctgga caccactagg ggtcagaagt agttcatcaa actttcttcc ctccctaatc
     1921 tcattggtta ccttgggcta tcgaaactta attaaccagt caagtcagct acttggcgag
     1981 atcgacttgt ctgggtttcg actacgctca gaattgcgtc agtcaagttc gatctggtcc
     2041 ttgctattgc acccgttctc cgattacgag tttcatttaa atcatgtgag caaaaggcca
     2101 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     2161 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     2221 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     2281 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     2341 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     2401 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     2461 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     2521 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     2581 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     2641 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2701 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     2761 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     2821 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     2881 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     2941 ctgtctattt cgttcatcca tagttgcatt taaatttccg aactctccaa ggccctcgtc
     3001 ggaaaatctt caaacctttc gtccgatcca tcttgcaggc tacctctcga acgaactatc
     3061 gcaagtctct tggccggcct tgcgccttgg ctattgcttg gcagcgccta tcgccaggta
     3121 ttactccaat cccgaatatc cgagatcggg atcacccgag agaagttcaa cctacatcct
     3181 caatcccgat ctatccgaga tccgaggaat atcgaaatcg gggcgcgcct ggtgtaccga
     3241 gaacgatcct ctcagtgcga gtctcgacga tccatatcgt tgcttggcag tcagccagtc
     3301 ggaatccagc ttgggaccca ggaagtccaa tcgtcagata ttgtactcaa gcctggtcac
     3361 ggcagcgtac cgatctgttt aaacctagat attgatagtc tgatcggtca acgtataatc
     3421 gagtcctagc ttttgcaaac atctatcaag agacaggatc agcaggaggc tttcgcatga
     3481 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     3541 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcgcgag
     3601 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     3661 aacgctttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     3721 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     3781 agtattcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     3841 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgattggag
     3901 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     3961 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     4021 tagcaatggc aacaaccttg cgtaaactat taactggcga actacttact ctagcttccc
     4081 ggcaacagtt gatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     4141 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     4201 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     4261 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     4321 tgattaagca ttggtaaccg attctaggtg cattggcgca gaaaaaaatg cctgatgcga
     4381 cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac
     4441 ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaagatc ccgaatcgtt
     4501 taaactcgac tctggctcta tcgaatctcc gtcgtttcga gcttacgcga acagccgtgg
     4561 cgctcatttg ctcgtcgggc atcgaatctc gtcagctatc gtcagcttac ctttttggca
     4621 gcgatcgcgg ctcccgacat cttggaccat tagctccaca ggtatcttct tccctctagt
     4681 ggtcataaca gcagcttcag ctacctctca attcaaaaaa cccctcaaga cccgtttaga
     4741 ggccccaagg ggttatgcta tcaatcgttg cgttacacac acaaaaaacc aacacacatc
     4801 catcttcgat ggatagcgat tttattatct aactgctgat cgagtgtagc cagatctagt
     4861 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
     4921 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     4981 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     5041 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     5101 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     5161 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg ctgatgcggt
     5221 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
     5281 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
     5341 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
     5401 atataagcag agctggttta gtgaaccgtc agatcagatc tttgtcgatc ctaccatcca
     5461 ctcgacacac ccgccagcgg ccgc
//
