LOCUS       sequence-295607-        7856 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 7856)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7856)
  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-m4134556
FEATURES             Location/Qualifiers
     source          1..7856
                     /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..1327
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    1396..1517
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1407..1426)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1461..1480
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1632..1678
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1708..1844
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1889..1960
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2186..2774)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2266..2285)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3274..4483
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4320..4339
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4487..5515
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5542..5590
                     /note="synthetic polyadenylation signal"
     CDS             join(5856..5924,5925..6716)
                     /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(6074..6093)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6729..6865
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7010..7081
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7086..7132)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7150..7198
                     /note="synthetic polyadenylation signal"
     enhancer        7279..7582
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7584..7786
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7737..7757
                     /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 cgaaagaaac cgcggcagct aagttcgaga ggcagcacat
     1321 ggacagttaa ggatccgcag gcctctgcta gcttgactga ctgagataca gcgtaccttc
     1381 agctcacaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     1441 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     1501 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     1561 aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatt ggcccatctc
     1621 tatcggtatc gtagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgtg
     1681 cccctcgggc cggattgcta tctaccggca ttggcgcaga aaaaaatgcc tgatgcgacg
     1741 ctgcgcgtct tatactccca catatgccag attcagcaac ggatacggct tccccaactt
     1801 gcccacttcc atacgtgtcc tccttaccag aaatttatcc ttaaggtcgt cagctatcct
     1861 gcaggcgatc tctcgatttc gatcaagaca ttcctttaat ggtcttttct ggacaccact
     1921 aggggtcaga agtagttcat caaactttct tccctcccta atctcattgg ttaccttggg
     1981 ctatcgaaac ttaattaacc agtcaagtca gctacttggc gagatcgact tgtctgggtt
     2041 tcgactacgc tcagaattgc gtcagtcaag ttcgatctgg tccttgctat tgcacccgtt
     2101 ctccgattac gagtttcatt taaatcatgt gagcaaaagg ccagcaaaag gccaggaacc
     2161 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     2221 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     2281 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     2341 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     2401 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     2461 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     2521 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     2581 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     2641 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     2701 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     2761 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     2821 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     2881 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     2941 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     3001 ccatagttgc atttaaattt ccgaactctc caaggccctc gtcggaaaat cttcaaacct
     3061 ttcgtccgat ccatcttgca ggctacctct cgaacgaact atcgcaagtc tcttggccgg
     3121 ccttgcgcct tggctattgc ttggcagcgc ctatcgccag gtattactcc aatcccgaat
     3181 atccgagatc gggatcaccc gagagaagtt caacctacat cctcaatccc gatctatccg
     3241 agatccgagg aatatcgaaa tcggggcgcg cctggcctcc gcgccgggtt ttggcgcctc
     3301 ccgcgggcgc ccccctcgtc acggcgagcg ctgccacgtc agacgaaggg cgcaggagcg
     3361 tcctgatcct tccgcccgga cgctcaggac agcggcccgc tgctcataag actcggcctt
     3421 agaaccccag tatcagcaga aggacatttt aggacgggac ttgggtgact ctagggcact
     3481 ggttttcttt ccagagagcg gaacaggcga ggaaaagtag tcccttctcg gcgattctgc
     3541 ggagggatct ccgtggggcg gtgaacgccg atgattatat aaggacgcgc cgggtgtggc
     3601 acagctagtt ccgtcgcagc cgggatttgg gtcgcggttc ttgtttgtgg atcgctgtga
     3661 tcgtcacttg gtgagtagcg ggctgctggg ctggccgggg ctttcgtggc cgccgggccg
     3721 ctcggtggga cggaagcgtg tggagagacc gccaagggct gtagtctggg tccgcgagca
     3781 aggttgccct gaactggggg ttggggggag cgcagcaaaa tggcggctgt tcccgagtct
     3841 tgaatggaag acgcttgtga ggcgggctgt gaggtcgttg aaacaaggtg gggggcatgg
     3901 tgggcggcaa gaacccaagg tcttgagccc ttcgctaatg cgggaaagct cttattcggg
     3961 tgagatgggc tgggcaccat ctggggaccc tgacgtgaag tttgtcactg actggagaac
     4021 tcggtttgtc gtctgttgcg ggggcggcag ttatggcggt gccgttgggc agtgcacccg
     4081 tacctttggg agcgcgcgcc ctcgtcgtgt cgtgacgtca cccgttctgt tggcttataa
     4141 tgcagggtgg ggccacctgc cggtaggtgt gcggtaggct tttctccgtc gcaggacgca
     4201 gggttcgggc ctagggtagg ctctcctgaa tcgacaggcg ccggacctct ggtgagggga
     4261 gggataagtg aggcgtcagt ttctttggtc ggttttatgt acctatcttc ttaagtagct
     4321 gaagctccgg ttttgaacta tgcgctcggg gttggcgagt gtgttttgtg aagtttttta
     4381 ggcacctttt gaaatgtaat catttgggtc aatatgtaat tttcagtgtt agacttgtaa
     4441 attgtccgct aaattctggc cgtttttggc ttttttgtta gacaacatgg gtaaaaagcc
     4501 tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga
     4561 cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg taggagggcg
     4621 tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc gttatgttta
     4681 tcggcacttt gcatccgccg cgctcccgat tccggaagtg cttgacattg gggagttcag
     4741 cgagagcctg acctattgca tctcccgccg tgcacagggt gtcacgttgc aagacctgcc
     4801 tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag gcaatggatg ccatcgctgc
     4861 cgccgatctt agccagacga gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata
     4921 cactacatgg cgtgatttca tatgcgcgat tgctgatccc catgtgtatc actggcaaac
     4981 tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg
     5041 ggccgaggac tgccccgaag tccggcacct cgtgcacgcg gatttcggct ccaacaatgt
     5101 cctgacggac aatggccgca taacagcggt cattgactgg agcgaggcga tgttcgggga
     5161 ttcccaatac gaggtcgcca acatcttctt ctggaggccg tggttggctt gtatggagca
     5221 gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgcccc ggctccgggc
     5281 gtatatgctc cgcattggtc ttgaccaact ctatcagagc ttggttgacg gcaatttcga
     5341 tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt
     5401 cgggcgtaca caaatcgccc gcagaagcgc cgccgtctgg accgatggct gtgtagaagt
     5461 actcgccgat agtggaaacc gacgccccag cactcgtccg agggcaaagg aataagctag
     5521 tatgtaagcc tagtcttaga taataaaatc gctatccatc gaagatggat gtgtgttggt
     5581 tttttgtgtg tgtaacgcta ggcgcgcctg gtgtaccgag aacgatcctc tcagtgcgag
     5641 tctcgacgat ccatatcgtt gcttggcagt cagccagtcg gaatccagct tgggacccag
     5701 gaagtccaat cgtcagatat tgtactcaag cctggtcacg gcagcgtacc gatctgttta
     5761 aacctagata ttgatagtct gatcggtcaa cgtataatcg agtcctagct tttgcaaaca
     5821 tctatcaaga gacaggatca gcaggaggct ttcgcatgag tattcaacat ttccgtgtcg
     5881 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     5941 tgaaagtaaa agatgctgaa gatcagttgg gtgcgcgagt gggttacatc gaactggatc
     6001 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgctttcca atgatgagca
     6061 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     6121 tcggtcgccg catacactat tctcagaatg acttggttga gtattcacca gtcacagaaa
     6181 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     6241 ataacactgc ggccaactta cttctgacaa cgattggagg accgaaggag ctaaccgctt
     6301 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     6361 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaaccttgc
     6421 gtaaactatt aactggcgaa ctacttactc tagcttcccg gcaacagttg atagactgga
     6481 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     6541 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     6601 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     6661 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaaccga
     6721 ttctaggtgc attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc
     6781 acatatgcca gattcagcaa cggatacggc ttccccaact tgcccacttc catacgtgtc
     6841 ctccttacca gaaatttatc cttaagatcg tttaaactcg actctggctc tatcgaatct
     6901 ccgtcgtttc gagcttacgc gaacagccgt ggcgctcatt tgctcgtcgg gcatcgaatc
     6961 tcgtcagcta tcgtcagctt acctttttgg cagcgatcgc ggctcccgac atcttggacc
     7021 attagctcca caggtatctt cttccctcta gtggtcataa cagcagcttc agctacctct
     7081 caattcaaaa aacccctcaa gacccgttta gaggccccaa ggggttatgc tatcaatcgt
     7141 tgcgttacac acacaaaaaa ccaacacaca tccatcttcg atggatagcg attttattat
     7201 ctaactgctg atcgagtgta gccagatcta gtaatcaatt acggggtcat tagttcatag
     7261 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     7321 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     7381 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     7441 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     7501 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     7561 attagtcatc gctattacca tgctgatgcg gttttggcag tacatcaatg ggcgtggata
     7621 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     7681 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
     7741 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg
     7801 tcagatcaga tctttgtcga tcctaccatc cactcgacac acccgccagc ggccgc
//
