LOCUS       sequence-307300-        7415 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 7415)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7415)
  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-m4133222
FEATURES             Location/Qualifiers
     source          1..7415
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    76..147
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(152..198)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    216..264
                     /note="synthetic polyadenylation signal"
     enhancer        345..648
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        650..852
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     803..823
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2184..2204
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2205..2246
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    2299..2420
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2310..2329)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2364..2383
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2535..2581
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2611..2747
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2792..2863
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3089..3677)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3169..3188)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4177..5386
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5223..5242
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5390..5989
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5390..5409)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5886..5906
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    6016..6064
                     /note="synthetic polyadenylation signal"
     CDS             join(6330..6398,6399..7190)
                     /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(6548..6567)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7203..7339
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 ctcatttgct cgtcgggcat cgaatctcgt cagctatcgt cagcttacct ttttggcagc
       61 gatcgcggct cccgacatct tggaccatta gctccacagg tatcttcttc cctctagtgg
      121 tcataacagc agcttcagct acctctcaat tcaaaaaacc cctcaagacc cgtttagagg
      181 ccccaagggg ttatgctatc aatcgttgcg ttacacacac aaaaaaccaa cacacatcca
      241 tcttcgatgg atagcgattt tattatctaa ctgctgatcg agtgtagcca gatctagtaa
      301 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
      361 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
      421 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
      481 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
      541 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
      601 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatgct gatgcggttt
      661 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
      721 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
      781 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
      841 ataagcagag ctggtttagt gaaccgtcag atcagatctt tgtcgatcct accatccact
      901 cgacacaccc gccagcggcc gccaccatgt ctgataatgg accccaaaat cagcgaaatg
      961 caccccgcat tacgtttggt ggaccctcag attcaactgg cagtaaccag aatggagaac
     1021 gcagtggggc gcgatcaaaa caacgtcggc cccaaggttt acccaataat actgcgtctt
     1081 ggttcaccgc tctcactcaa catggcaagg aagaccttaa attccctcga ggacaaggcg
     1141 ttccaattaa caccaatagc agtccagatg accaaattgg ctactaccga agagctacca
     1201 gacgaattcg tggtggtgac ggtaaaatga aagatctcag tccaagatgg tatttctact
     1261 acctaggaac tgggccagaa gctggacttc cctatggtgc taacaaagac ggcatcatat
     1321 gggttgcaac tgagggagcc ttgaatacac caaaagatca cattggcacc cgcaatcctg
     1381 ctaacaatgc tgcaatcgtg ctacaacttc ctcaaggaac aacattgcca aaaggcttct
     1441 acgcagaagg gagcagaggc ggcagtcaag cctcttctcg ttcctcatca cgtagtcgca
     1501 acagttcaag aaattcaact ccaggcagca gtaggggaac ttctcctgct agaatggctg
     1561 gcaatggcgg tgatgctgct cttgctttgc tgctgcttga cagattgaac cagcttgaga
     1621 gcaaaatgtc tggtaaaggc caacaacaac aaggccaaac tgtcactaag aaatctgctg
     1681 ctgaggcttc taagaagcct cggcaaaaac gtactgccac taaagcatac aatgtaacac
     1741 aagctttcgg cagacgtggt ccagaacaaa cccaaggaaa ttttggggac caggaactaa
     1801 tcagacaagg aactgattac aaacattggc cgcaaattgc acaatttgcc cccagcgctt
     1861 cagcgttctt cggaatgtcg cgcattggca tggaagtcac accttcggga acgtggttga
     1921 cctacacagg tgccatcaaa ttggatgaca aagatccaaa tttcaaagat caagtcattt
     1981 tgctgaataa gcatattgac gcatacaaaa cattcccacc aacagagcct aaaaaggaca
     2041 aaaagaagaa ggctgatgaa actcaagcct taccgcagag acagaagaaa cagcaaactg
     2101 tgactcttct tcctgctgca gatttggatg atttctccaa acaattgcaa caatccatga
     2161 gcagtgctga ctcaactcag gccgaaaacc tttactttca gtcggggaaa cctataccca
     2221 atccacttct cggcttagat tctacctaag ctagcttgac tgactgagat acagcgtacc
     2281 ttcagctcac agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     2341 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     2401 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     2461 gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt attggcccat
     2521 ctctatcggt atcgtagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
     2581 gtgcccctcg ggccggattg ctatctaccg gcattggcgc agaaaaaaat gcctgatgcg
     2641 acgctgcgcg tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa
     2701 cttgcccact tccatacgtg tcctccttac cagaaattta tccttaaggt cgtcagctat
     2761 cctgcaggcg atctctcgat ttcgatcaag acattccttt aatggtcttt tctggacacc
     2821 actaggggtc agaagtagtt catcaaactt tcttccctcc ctaatctcat tggttacctt
     2881 gggctatcga aacttaatta accagtcaag tcagctactt ggcgagatcg acttgtctgg
     2941 gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc tggtccttgc tattgcaccc
     3001 gttctccgat tacgagtttc atttaaatca tgtgagcaaa aggccagcaa aaggccagga
     3061 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3121 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3181 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3241 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3301 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3361 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3421 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3481 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     3541 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     3601 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     3661 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     3721 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     3781 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     3841 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     3901 catccatagt tgcatttaaa tttccgaact ctccaaggcc ctcgtcggaa aatcttcaaa
     3961 cctttcgtcc gatccatctt gcaggctacc tctcgaacga actatcgcaa gtctcttggc
     4021 cggccttgcg ccttggctat tgcttggcag cgcctatcgc caggtattac tccaatcccg
     4081 aatatccgag atcgggatca cccgagagaa gttcaaccta catcctcaat cccgatctat
     4141 ccgagatccg aggaatatcg aaatcggggc gcgcctggcc tccgcgccgg gttttggcgc
     4201 ctcccgcggg cgcccccctc gtcacggcga gcgctgccac gtcagacgaa gggcgcagga
     4261 gcgtcctgat ccttccgccc ggacgctcag gacagcggcc cgctgctcat aagactcggc
     4321 cttagaaccc cagtatcagc agaaggacat tttaggacgg gacttgggtg actctagggc
     4381 actggttttc tttccagaga gcggaacagg cgaggaaaag tagtcccttc tcggcgattc
     4441 tgcggaggga tctccgtggg gcggtgaacg ccgatgatta tataaggacg cgccgggtgt
     4501 ggcacagcta gttccgtcgc agccgggatt tgggtcgcgg ttcttgtttg tggatcgctg
     4561 tgatcgtcac ttggtgagta gcgggctgct gggctggccg gggctttcgt ggccgccggg
     4621 ccgctcggtg ggacggaagc gtgtggagag accgccaagg gctgtagtct gggtccgcga
     4681 gcaaggttgc cctgaactgg gggttggggg gagcgcagca aaatggcggc tgttcccgag
     4741 tcttgaatgg aagacgcttg tgaggcgggc tgtgaggtcg ttgaaacaag gtggggggca
     4801 tggtgggcgg caagaaccca aggtcttgag cccttcgcta atgcgggaaa gctcttattc
     4861 gggtgagatg ggctgggcac catctgggga ccctgacgtg aagtttgtca ctgactggag
     4921 aactcggttt gtcgtctgtt gcgggggcgg cagttatggc ggtgccgttg ggcagtgcac
     4981 ccgtaccttt gggagcgcgc gccctcgtcg tgtcgtgacg tcacccgttc tgttggctta
     5041 taatgcaggg tggggccacc tgccggtagg tgtgcggtag gcttttctcc gtcgcaggac
     5101 gcagggttcg ggcctagggt aggctctcct gaatcgacag gcgccggacc tctggtgagg
     5161 ggagggataa gtgaggcgtc agtttctttg gtcggtttta tgtacctatc ttcttaagta
     5221 gctgaagctc cggttttgaa ctatgcgctc ggggttggcg agtgtgtttt gtgaagtttt
     5281 ttaggcacct tttgaaatgt aatcatttgg gtcaatatgt aattttcagt gttagacttg
     5341 taaattgtcc gctaaattct ggccgttttt ggcttttttg ttagacaaca tgaccgagta
     5401 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc
     5461 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga
     5521 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt
     5581 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc
     5641 gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc
     5701 cgcgcagcaa cagatggagg gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt
     5761 cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt
     5821 gctccccgga gtggaggccg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc
     5881 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt
     5941 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctagg ctagtatgta
     6001 agcctagtct tagataataa aatcgctatc catcgaagat ggatgtgtgt tggttttttg
     6061 tgtgtgtaac gctaggcgcg cctggtgtac cgagaacgat cctctcagtg cgagtctcga
     6121 cgatccatat cgttgcttgg cagtcagcca gtcggaatcc agcttgggac ccaggaagtc
     6181 caatcgtcag atattgtact caagcctggt cacggcagcg taccgatctg tttaaaccta
     6241 gatattgata gtctgatcgg tcaacgtata atcgagtcct agcttttgca aacatctatc
     6301 aagagacagg atcagcagga ggctttcgca tgagtattca acatttccgt gtcgccctta
     6361 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     6421 taaaagatgc tgaagatcag ttgggtgcgc gagtgggtta catcgaactg gatctcaaca
     6481 gcggtaagat ccttgagagt tttcgccccg aagaacgctt tccaatgatg agcactttta
     6541 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     6601 gccgcataca ctattctcag aatgacttgg ttgagtattc accagtcaca gaaaagcatc
     6661 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     6721 ctgcggccaa cttacttctg acaacgattg gaggaccgaa ggagctaacc gcttttttgc
     6781 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     6841 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacc ttgcgtaaac
     6901 tattaactgg cgaactactt actctagctt cccggcaaca gttgatagac tggatggagg
     6961 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     7021 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     7081 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     7141 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ccgattctag
     7201 gtgcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat
     7261 gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt
     7321 accagaaatt tatccttaag atcccgaatc gtttaaactc gactctggct ctatcgaatc
     7381 tccgtcgttt cgagcttacg cgaacagccg tggcg
//
