LOCUS       sequence-295629-        7408 bp    DNA     circular SYN 15-APR-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 7408)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7408)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGBW-m4134550
FEATURES             Location/Qualifiers
     source          1..7408
                     /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..1324
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    1377..1498
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1388..1407)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1442..1461
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1613..1659
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1689..1825
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1870..1941
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2167..2755)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2247..2266)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3255..4464
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4301..4320
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4468..5067
                     /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(4468..4487)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4964..4984
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5094..5142
                     /note="synthetic polyadenylation signal"
     CDS             join(5408..5476,5477..6268)
                     /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(5626..5645)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6281..6417
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6562..6633
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6638..6684)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6702..6750
                     /note="synthetic polyadenylation signal"
     enhancer        6831..7134
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7136..7338
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7289..7309
                     /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 cggggaaacc tatacccaat ccacttctcg gcttagattc
     1321 tacctaagct agcttgactg actgagatac agcgtacctt cagctcacag acatgataag
     1381 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     1441 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     1501 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
     1561 aagcaagtaa aacctctaca aatgtggtat tggcccatct ctatcggtat cgtagcataa
     1621 ccccttgggg cctctaaacg ggtcttgagg ggttttttgt gcccctcggg ccggattgct
     1681 atctaccggc attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc
     1741 acatatgcca gattcagcaa cggatacggc ttccccaact tgcccacttc catacgtgtc
     1801 ctccttacca gaaatttatc cttaaggtcg tcagctatcc tgcaggcgat ctctcgattt
     1861 cgatcaagac attcctttaa tggtcttttc tggacaccac taggggtcag aagtagttca
     1921 tcaaactttc ttccctccct aatctcattg gttaccttgg gctatcgaaa cttaattaac
     1981 cagtcaagtc agctacttgg cgagatcgac ttgtctgggt ttcgactacg ctcagaattg
     2041 cgtcagtcaa gttcgatctg gtccttgcta ttgcacccgt tctccgatta cgagtttcat
     2101 ttaaatcatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     2161 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     2221 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     2281 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     2341 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     2401 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     2461 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     2521 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     2581 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     2641 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     2701 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     2761 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     2821 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     2881 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     2941 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg catttaaatt
     3001 tccgaactct ccaaggccct cgtcggaaaa tcttcaaacc tttcgtccga tccatcttgc
     3061 aggctacctc tcgaacgaac tatcgcaagt ctcttggccg gccttgcgcc ttggctattg
     3121 cttggcagcg cctatcgcca ggtattactc caatcccgaa tatccgagat cgggatcacc
     3181 cgagagaagt tcaacctaca tcctcaatcc cgatctatcc gagatccgag gaatatcgaa
     3241 atcggggcgc gcctggcctc cgcgccgggt tttggcgcct cccgcgggcg cccccctcgt
     3301 cacggcgagc gctgccacgt cagacgaagg gcgcaggagc gtcctgatcc ttccgcccgg
     3361 acgctcagga cagcggcccg ctgctcataa gactcggcct tagaacccca gtatcagcag
     3421 aaggacattt taggacggga cttgggtgac tctagggcac tggttttctt tccagagagc
     3481 ggaacaggcg aggaaaagta gtcccttctc ggcgattctg cggagggatc tccgtggggc
     3541 ggtgaacgcc gatgattata taaggacgcg ccgggtgtgg cacagctagt tccgtcgcag
     3601 ccgggatttg ggtcgcggtt cttgtttgtg gatcgctgtg atcgtcactt ggtgagtagc
     3661 gggctgctgg gctggccggg gctttcgtgg ccgccgggcc gctcggtggg acggaagcgt
     3721 gtggagagac cgccaagggc tgtagtctgg gtccgcgagc aaggttgccc tgaactgggg
     3781 gttgggggga gcgcagcaaa atggcggctg ttcccgagtc ttgaatggaa gacgcttgtg
     3841 aggcgggctg tgaggtcgtt gaaacaaggt ggggggcatg gtgggcggca agaacccaag
     3901 gtcttgagcc cttcgctaat gcgggaaagc tcttattcgg gtgagatggg ctgggcacca
     3961 tctggggacc ctgacgtgaa gtttgtcact gactggagaa ctcggtttgt cgtctgttgc
     4021 gggggcggca gttatggcgg tgccgttggg cagtgcaccc gtacctttgg gagcgcgcgc
     4081 cctcgtcgtg tcgtgacgtc acccgttctg ttggcttata atgcagggtg gggccacctg
     4141 ccggtaggtg tgcggtaggc ttttctccgt cgcaggacgc agggttcggg cctagggtag
     4201 gctctcctga atcgacaggc gccggacctc tggtgagggg agggataagt gaggcgtcag
     4261 tttctttggt cggttttatg tacctatctt cttaagtagc tgaagctccg gttttgaact
     4321 atgcgctcgg ggttggcgag tgtgttttgt gaagtttttt aggcaccttt tgaaatgtaa
     4381 tcatttgggt caatatgtaa ttttcagtgt tagacttgta aattgtccgc taaattctgg
     4441 ccgtttttgg cttttttgtt agacaacatg accgagtaca agcccacggt gcgcctcgcc
     4501 acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc
     4561 gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa
     4621 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc
     4681 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc
     4741 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggagggc
     4801 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg
     4861 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggccgcc
     4921 gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac
     4981 gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg
     5041 tgcatgaccc gcaagcccgg tgcctaggct agtatgtaag cctagtctta gataataaaa
     5101 tcgctatcca tcgaagatgg atgtgtgttg gttttttgtg tgtgtaacgc taggcgcgcc
     5161 tggtgtaccg agaacgatcc tctcagtgcg agtctcgacg atccatatcg ttgcttggca
     5221 gtcagccagt cggaatccag cttgggaccc aggaagtcca atcgtcagat attgtactca
     5281 agcctggtca cggcagcgta ccgatctgtt taaacctaga tattgatagt ctgatcggtc
     5341 aacgtataat cgagtcctag cttttgcaaa catctatcaa gagacaggat cagcaggagg
     5401 ctttcgcatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     5461 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     5521 gggtgcgcga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     5581 tcgccccgaa gaacgctttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     5641 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     5701 tgacttggtt gagtattcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     5761 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     5821 aacgattgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     5881 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     5941 cacgatgcct gtagcaatgg caacaacctt gcgtaaacta ttaactggcg aactacttac
     6001 tctagcttcc cggcaacagt tgatagactg gatggaggcg gataaagttg caggaccact
     6061 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     6121 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     6181 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     6241 aggtgcctca ctgattaagc attggtaacc gattctaggt gcattggcgc agaaaaaaat
     6301 gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc aacggatacg
     6361 gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta tccttaagat
     6421 cgtttaaact cgactctggc tctatcgaat ctccgtcgtt tcgagcttac gcgaacagcc
     6481 gtggcgctca tttgctcgtc gggcatcgaa tctcgtcagc tatcgtcagc ttaccttttt
     6541 ggcagcgatc gcggctcccg acatcttgga ccattagctc cacaggtatc ttcttccctc
     6601 tagtggtcat aacagcagct tcagctacct ctcaattcaa aaaacccctc aagacccgtt
     6661 tagaggcccc aaggggttat gctatcaatc gttgcgttac acacacaaaa aaccaacaca
     6721 catccatctt cgatggatag cgattttatt atctaactgc tgatcgagtg tagccagatc
     6781 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     6841 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     6901 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     6961 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     7021 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     7081 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catgctgatg
     7141 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
     7201 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
     7261 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
     7321 gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc gatcctacca
     7381 tccactcgac acacccgcca gcggccgc
//
